B Y T

TRNG I HC DC H NI

TRN TH BCH NGC

NGHIN CU TC DNG H TR

IU TR GT TRN THC NGHIM

CA HT CN TY

(APIUM GRAVEOLENS L., APIACEAE)

KHA LUN TT NGHIP DC S

H NI 2014

B Y T

TRNG I HC DC H NI

TRN TH BCH NGC

NGHIN CU TC DNG H TR

IU TR GT TRN THC NGHIM

CA HT CN TY

(APIUM GRAVEOLENS L., APIACEAE)

KHA LUN TT NGHIP DC S

Ngi hng dn:

TS. Nguyn Thy Dng

DS. V Th Phng Tho

Ni thc hin:

B mn Dc lc

H NI - 2014

LI CM N

hon thnh kha lun ny, ti nhn c s gip rt nhiu ca

thy c, bn b v ngi thn.

Trc tin, ti xin by t lng bit n su sc ti TS. Nguyn Thy Dng

v DS. V Th Phng Tho, nhng ngi thy lun quan tm, gip , hng

dn v ng vin ti trong sut qu trnh hon thnh ti ny.

Xin chn thnh cm n cc thy, c v anh ch k thut vin B mn Dc

lc, gip v to iu kin cho ti trong sut qu trnh nghin cu v thc

hin ti.

Xin chn thnh cm n cc cn b phng o to, cc b mn, phng ban

khc ca trng i hc Dc H Ni.

Li cui cng, ti xin gi li cm n n gia nh v bn b, nhng ngi

lun ng vin, gip v ng gp kin cho ti hon thnh kha lun ny.

H Ni, thng 5, nm 2014.

Sinh vin

Trn Th Bch Ngc

MC LC

LI CM N

DANH MC CC BNG

DANH MC CC HNH

DANH MC CC K HIU, CH VIT TT

T VN ............................................................................................................. 1

CHNG 1: TNG QUAN ....................................................................................... 2

1.1. I CNG V BNH GT ............................................................................ 2

1.1.1. nh ngha ......................................................................................................... 2

1.1.2. Qu trnh sinh tng hp acid uric ..................................................................... 2

1.1.3. C ch bnh sinh ca gt .................................................................................. 3

1.1.4. C ch gy vim cp ca tinh th urat .............................................................. 3

1.1.5. Thuc s dng trong iu tr gt v tng acid uric mu .................................. 4

1.1.5.1. Thuc chng vim ................................................................................... 4

1.1.5.2. Thuc gim tng hp acid uric..6

1.1.5.3. Thuc gy tng thi acid uric ................................................................. 7

1.1.5.4. Thuc tiu acid uric ................................................................................ 8

1.2. TNG QUAN V CY CN TY .................................................................... 8

1.2.1. Tn gi ............................................................................................................... 8

1.2.2. c im thc vt ............................................................................................. 9

1.2.3. Phn b ........................................................................................................... 10

1.2.4. Thnh phn ha hc ca ht cn ty .............................................................. 10

1.2.5. Tc dng sinh hc ca ht cn ty .................................................................. 13

1.2.6. Cng dng ca ht cn ty.............................................................................. 15

CHNG 2: I TNG V PHNG PHP NGHIN CU ........................ 16

2.1. NGUYN LIU NGHIN CU ...................................................................... 16

2.1.1. Dc liu nghin cu ...................................................................................... 16

2.1.2. Chun b dch chit dc liu ......................................................................... 16

2.2. PHNG TIN NGHIN CU ....................................................................... 17

2.2.1. ng vt th nghim ........................................................................................ 17

2.2.2. Ha cht, thuc th ......................................................................................... 17

2.2.3. Thit b ............................................................................................................ 18

2.3. NI DUNG NGHIN CU .............................................................................. 19

2.4. PHNG PHP NGHIN CU ...................................................................... 20

2.4.1. Phng php nh gi tc dng h acid uric huyt thanh trn m hnh gy

tng acid uric cp thc nghim. ................................................................................ 20

2.4.2. Phng php nh gi tc dng c ch xanthin oxidase in vitro v in vivo .. 21

2.4.3. Phng php nh gi tc dng chng vim thc nghim ............................. 24

2.4.3.1. nh gi tc dng chng vim cp trn m hnh gy ph bn chn

chut bng carrageenan .................................................................................... 24

2.4.3.2.Phng php nh gi tc dng chng vim trn m hnh gy vim

mng hot dch khp gi ................................................................................... 25

2.4.4. Phng php x l s liu ............................................................................... 26

CHNG 3: KT QU ........................................................................................... 27

3.1. KT QU NH GI TC DNG H ACID URIC CA CAO TON

PHN HT CN TY ............................................................................................ 27

3.2. KT QU NH GI TC DNG C CH XO IN VITRO V IN VIVO

CA CAO TON PHN HT CN TY ............................................................ 29

3.2.1. Kt qu nh gi tc dng c ch XO in vitro ca cao ton phn ht cn

ty29

3.2.2. Kt qu nh gi tc dng c ch XO in vivo ca cao ton phn ht cn

ty ............................................................................................................ 30

3.3. KT QU NH GI TC DNG CHNG VIM CA CAO TON

PHN HT CN TY ........................................................................................... 32

3.3.1. Kt qu nh gi tc dng chng vim trn m hnh gy ph bn chn chut

bng carrageenan ..................................................................................................... 32

3.3.2. Kt qu nh gi tc dng chng vim trn m hnh gy vim mng hot dch

khp gi ................................................................................................................. 34

CHNG 4: BN LUN ........................................................................................ 37

4.1. V TC DNG H ACID URIC CA CAO TON PHN HT CN TY

... .................................................................................................................... 37

4.2. V TC DNG C CH XANTHIN OXIDASE CA CAO TON PHN

HT CN TY ........................................................................................................ 38

4.3. V TC DNG CHNG VIM CA CAO TON PHN HT CN

TY. .................................................................................................. 41

KT LUN V XUT ...................................................................................... 44

TI LIU THAM KHO

DANH MC CC BNG

STT K hiu Tn bng Trang

1 1.1 Cc hp cht flavonoid c trong ht cn ty 11

2 2.1 Thnh phn mu chng v mu th 21

3 2.2 Thang im nh gi mc vim da trn triu

chng 26

4 3.1

Kt qu nh gi tc dng h acid uric ca cao

ton phn ht cn ty trn m hnh gy tng acid

uric cp

28

5 3.2 Kt qu nh gi tc dng c ch XO in vitro ca

cao ton phn ht cn ty 29

6 3.3 Kt qu nh gi tc dng c ch XO in vivo ca

cao ton phn ht cn ty 31

7 3.4 Kt qu nh gi tc dng chng vim trn m

hnh gy ph bn chn chut bng carrageenan 33

8 3.5

Kt qu nh gi tc dng chng vim trn m

hnh gy vim mng hot dch khp gi bng tinh

th natri urat

35

DANH MC CC HNH

STT K hiu Tn hnh Trang

1 1.1 Qa trnh hnh thnh acid uric trong c th 2

2 1.2 C ch gy vim ca tinh th urat 4

3 1.3 C ch chng vim ca NSAIDs v glucocorticoid 6

4 1.4 Cy cn ty (Apium graveolens L.) 9

5 1.5

Khung cu trc ca cc hp cht flavonoid c trong

ht cn ty 11

6 2.1 Quy trnh chit xut cao ton phn ht cn ty 17

7 2.2 Thit k nghin cu 19

8 2.3 Quy trnh th nghim nh gi tc dng h acid uric

mu trn m hnh gy tng cp bng kali oxonat 20

9 2.4 Quy trnh th nghim nh gi tc dng c ch XO

in vitro 22

10 2.5 Quy trnh nh gi tc dng chng vim trn m

hnh gy ph bn chn chut bng carrageenan 24

11 3.1 Kt qu nh gi tc dng h acid uric ca cao ton

phn ht cn ty trn m hnh gy tng acid uric cp 28

12 3.2 Kt qu nh gi tc dng c ch XO in vivo ca

cao ton phn ht cn ty 34

13 3.3 Kt qu nh gi tc dng chng vim trn m hnh

gy ph bn chn chut bng carrageenan 35

DANH MC CC K HIU, CH VIT TT

XO : xanthin oxidase

NSAIDs : non steroidal anti inflammatory drug

1

T VN

Gt l mt bnh ri lon chuyn ha vi biu hin lm sng l tng acid uric

mu. Ti Vit Nam, vi s pht trin ca kinh t x hi cng nh ch dinh

dng thiu hp l, bnh gt ngy cng tr nn ph bin hn [3]. Bnh c th cp

v mn tnh u c nhng triu chng in hnh nh vim, au d di, kh vn

ng lm nh hng khng nh ti cht lng cuc sng ca bnh nhn. Vic s

dng cc thuc ha dc gim au, chng vim, h acid uric mu mc d cho hiu

qu cao song vn cn hn ch v tc dng khng mong mun v gi thnh, nht l

khi bnh nhn phi dng thuc lu di.

Vit Nam vi ngun dc liu a dng v phong ph c nhiu cy thuc

c s dng iu tr bnh gt. Mt s nghin cu c thc hin nhm tm

kim nhng dc liu mi c kh nng iu tr gt, pht hin nhiu dc liu c

kh nng c ch hot ng ca enzym xanthin oxidase in vitro, trong c cy cn

ty (Apium graveolens L., h Cn, Apiaceae) [19], [25]. Mt khc, trong y hc c

truyn, cn ty c dng iu tr bnh thng phong (bnh gt theo y hc c

truyn) v cc bnh lin quan v khp nh thp khp, au khp, vim khp [8],

[13], [20]. pht trin dc liu ny thnh ch phm dng trong iu tr gt, cn

nh gi tip tc dng ca cy cn ty trn ng vt thc nghim. Vit Nam hin

cha c nghin cu no c trin khai nh gi tc dng iu tr gt ca cy

cn ty. Vi mong mun gp phn iu tr v d phng ti pht bnh gt bng

thuc c ngun gc dc liu da trn c s khoa hc, ti Nghin cu tc

dng h tr iu tr gt trn thc nghim ca ht cn ty (Apium graveolens

L., Apiaceae) c thc hin vi ba mc tiu:

1. nh gi tc dng h acid uric ca ht cn ty trn thc nghim.

2. nh gi tc dng c ch xanthin oxidase in vitro v in vivo ca ht

cn ty.

3. nh gi tc dng chng vim ca ht cn ty trn thc nghim.

2

CHNG 1: TNG QUAN

1.1. I CNG V BNH GT

1.1.1. nh ngha

Gt l bnh ri lon chuyn ha acid uric, c c im chnh l tng acid uric

mu. Khi acid uric bo ha dch ngoi bo s gy lng ng cc tinh th

monodium urat cc m. Ty theo v tr tinh th urat b tch ly m bnh biu hin

bi mt hay nhiu triu chng lm sng: vim khp v cnh khp cp v/hoc mn

tnh, thng c gi l vim khp do gt; ht tophi m mm; bnh thn do gt

hoc si tit niu [3].

1.1.2. Qu trnh sinh tng hp acid uric

cid uric l sn phm ca qu trnh thoi ging cc nucleo-protein c cha

nhn purin adenin, guanin trong c th vi s tham gia ca cc enzym:

phosphoribosyl pyrophosphatase (PRPP), nucleoside phosphorylase, xanthin

oxidase (XO) [15] hnh1.1 .

Hnh 1.1. Qu trnh hnh thnh acid uric trong c th [15]

3

1.1.3. C ch bnh sinh ca gt

C ch bnh sinh chnh ca gt l s tng acid uric mu. Tng acid uric c

nh ngha l khi nng acid uric mu vt qu gii hn ha tan ti a ca acid

uric huyt tng: nam trn 7,0 mg/dL tc trn 420 mol/L ; n trn 6,0 mg/dL (tc

trn 360 mol/L [12]. Trong mu, lng acid uric thng t n mc ha tan

nht nh [50]. Hng ngy c khong 700mg acid uric c to ra bao gm ni sinh

(600mg) v ngoi sinh 100mg . Trong , 30% b phn hy bi vi khun ng

rut, cn li 70% c bi tit qua thn [16]. Khi nng acid uric trong huyt

thanh cao hn nng bo ha, dn ti tch ly tinh th urat ti cc m, to nn cc

ht microtophi. S lng ng ca cc ht ny ti cc v tr khc nhau s gy nn cc

tn thng khc nhau. Ti sn khp, khi cc ht ny v ra, s khi pht cn gt

cp; s lng ng ca cc vi tinh th cnh khp, trong mng hot dch, trong m

sn v m xng s dn n bnh xng khp mn tnh do gt; s c mt vi tinh

th urat ti m mm, bao gn to nn ht tophi; v cui cng, lng ng ti t chc

k ca thn gy vim thn k [3].

Nh vy, vic tc ng vo cc yu t tham gia vo qu trnh sinh tng hp v

chuyn ha acid uric s l mt trong cc cch lm thay i nng acid uric mu.

1.1.4. C ch gy vim cp ca tinh th urat

Trong bnh gt, phn ng vim l nguyn nhn chnh gy ra triu chng au d

di bnh nhn. Cc nghin cu gn y xc nh c c ch gy vim do tinh

th urat bnh gt. Phn ng vim chia lm 2 giai on [29], [34]:

- Giai on 1: Sn xut ra IL-1 interleukin-1 . u tin, bch cu n nhn

trung tnh thc bo cc tinh th urat, dn ti mt lot cc qu trnh truyn tn

hiu thng qua cc cht truyn tin MyD88 (l protein p ng t bo ty bit

ha s cp 88), NALPT (Nacht, Leucin rich repeat v PYD-containing protein

3) l mt loi protein gy vim, lm hot ha dng tin IL-1 thnh IL-1 hot

ha nh enzym caspase-1.

4

- Giai on 2: Khi c tn hiu ca IL-1, IL-1R v TIR-MyD88 (TIR hay

Toll/IL-1 c t bo khng ngun gc ty xng , MyD88 c hot ha.

Do m cc chemokin v cc cht trung gian gy vim c gii phng.

ng thi chnh cc cht ny kch thch sn sinh ra IL-1, TNF- tip tc gy

vim trm trng hn.

C ch gy vim c m t hnh 1.2.

Hnh 1.2. C ch gy vim ca tinh th urat [3]

1.1.5. Thuc s dng trong iu tr gt v tng acid uric mu

1.1.5.1. Thuc chng vim

NSAIDs

5

Tc dng: Gim au mc nh v trung bnh, c tc dng tt i vi cc loi

au do vim. Thuc c tc dng chng vim trn hu ht cc loi vim, khng k

nguyn nhn.

C ch: Cc thuc ny c tc dng chng vim do c ch enzym

cyclooxygenase COX , ngn cn tng hp prostagrandin l cht trung gian ha hc

gy vim, do lm gim qu trnh vim. Ngoi ra, thuc cn ngn cn qu trnh

bin i protein, do lm vng bn mng lysosom v i khng vi tc dng ca

cc cht trung gian ha hc nh: bradykinin, histamin, serotonin...[2] c ch c

th hin hnh 1.3).

Ch nh: iu tr cn gt cp tnh [2].

Tc dng khng mong mun: trn tiu ha (lot d dy), trn mu (chy mu),

trn thn (vim thn k) [2].

Corticoid

Tc dng: Thuc c tc dng chng vim mnh, do mi nguyn nhn c hc,

min dch v nhim khun). Do c ch phospholipase C nn thuc lm gim gii

phng histamin v cc cht trung gian ha hc gy d ng. c bit, glucocorticoid

c ch min dch do gim s lng t bo lympho, c ch chc nng thc bo v

sn xut khng th. Ngoi ra, thuc cn c tc dng trn chuyn ha (glucid, lipid,

protid, chuyn ha mui nc) [2].

C ch chng vim: Glucocorticoid c ch phospholipase A2 thng qua kch

thch tng hp lipocortin, lm gim tng hp leukotrien v prostaglandin. Ngoi ra

thuc cn c tc dng c ch dng bch cu n nhn, a nhn lympho bo i vo

m gy khi pht vim [2] c ch c th hin hnh 1.3).

Ch nh: iu tr gt chng ch nh vi NSAIDs v colchicin [17], [31].

Tc dng khng mong mun: Glucocorticoid thng gy long xng, lot d

dy t trng, ph, tng huyt p v cc ri lon khc [2].

Colchicin

Cochicin l alkaloid ca cy Cholchicum autumnale c s dng cho bnh

au khp t th k 16.

6

Tc dng: Thuc c tc dng tt trong au do gt, khng c tc dng trong

trng hp khng phi bnh gt [2].

C ch: Colchicin lm gim s di chuyn ca bch cu, gim s tp trung ca

bch cu ht t chc vim, c ch thc bo cc tinh th urat v do km hm sn

xut lactic, gi cho pH ti ch c bnh thng [2], [17].

Ch nh: iu tr gt cp [3].

Tc dng khng mong mun: Colchicin c tc dng nhanh, mnh nhng

thng gp bun nn, a chy v dit ty xng [31], [38].

Hnh 1.3. C ch chng vim ca NSAIDs v glucocorticoid

1.1.5.2. Thuc gim tng hp acid uric

Allopurinol

Tc dng: Thuc lm gim nng acid uric mu. Ngoi ra, thuc lm tng

bi xut cc tin cht ca acid uric qua nc tiu [2].

7

C ch: Allopurinol c ch enzym xanthin oxidase, l enzym c vai tr

chuyn tin cht hypoxanthin v xanthin thnh acid uric, nh lm gim nng

acid uric mu [2].

Ch nh: iu tr gt mn [1].

Tc dng khng mong mun: Thng gp kch ng tiu ha, c vi gan [10].

Febuxostat

Fubuxostat l thuc mi c a vo th trng trong nhng nm gn y

nh mt thuc thay th allopurinol [29].

Tc dng v c ch: Thuc c tc dng h nng acid uric mu theo c ch

c ch enzym xanthin oxidase, tng t allopurinol [29].

Ch nh: iu tr gt khi c vng da , tn thng. c bit l d phng khi

khng c colchicin [38].

Tc dng khng mong mun: Tc dng khng mong mun thng gp nht

khi s dng febuxostat l lm bt thng chc nng gan, tiu chy, au u, bun

nn, pht ban, cn gt cp [38].

1.1.5.3. Thuc gy tng thi acid uric

Probenecid

Tc dng: Thuc c tc dng ln s bi tit acid uric qua nc tiu theo c hai

chiu. liu iu tr thuc lm tng thi acid uric, liu thp thuc c tc dng

ngc li [33].

C ch: Probenecid c ch cnh tranh h vn chuyn anion - urat gy c ch

ti hp thu acid uric, gy tng thi tr acid uric ng thn [2].

Ch nh: iu tr gt mn [2].

Tc dng khng mong mun: Probenecid c th gy si tit niu,. V vy

thng trnh dng thuc cho nhng bnh nhn c tin s si tit niu [21].

Sulfinpyrazon

Tc dng v c ch: Sulfinpyrazon c c ch v tc dng thi tr urat tng t

nhng hiu qu cao hn probenecid. Tc dng ca thuc ph thuc vo liu dng

tng t probenecid [50].

8

Ch nh: Phi hp vi colchicin hoc NS IDS d phng cn gt cp.

Tc dng khng mong mun: C th gp au bng, bun nn, nn, lot d dy

t trng hoc tai bin trn mu ging nh phenylbutazon [7].

Ngoi ra, trong nhm thuc ny cn c: benzbromaron, fenofibrat v losartan.

1.1.5.4. Thuc tiu acid uric

Trong c th hu ht cc loi ng vt c v u c enzym uricase, enzym ny

c vai tr chuyn urat thnh allantoin d tan v d bi tit hn, v th chng c nng

acid urat mu thp v khng b gt. Nm 1957, London v Hudson xc nh rng

dng uricase tinh khit tim tnh mch c tc dng phn gii acid uric [6].

Rasburicase

Tc dng v c ch: Thuc c tc dng h nng urat mu. Rasburicase xc

tc phn ng oxi ha ca acid uric thnh cht chuyn ha allantoin khng hot tnh,

sn phm chuyn ha ha tan nhiu trong nc hn acid uric.

Ch nh: FDA gi c th dng rasburicase trn ngi ln (tuy nhin c

khuyn co hn ch). Thuc c thi gian bn thi ngn, dung np km, tnh khng

nguyn cao nn cha c p dng iu tr gt [7].

Pegylat uricase

Pegylat uricase l mt dng uricase ci tin, c gn thm chui polyethylen

glycol nhm lm gim tnh khng nguyn v ko di thi gian bn thi ca uricase.

Cc th nghim lm sng giai on I v II cho thy thuc c hiu qu h acid uric

mu, dung np tt. Hin nay, thuc vn ang trong giai on th nghim lm sng

[7].

1.2. TNG QUAN V CY CN TY

1.2.1. Tn gi

- Tn khoa hc: Apium graveolens L. h Cn (Apiaceae) [9], [11], [13].

- Tn nc ngoi: Celery ( nh ; celeri, prersildes marais, ache des marais

(Php) [13].

- Tn thng gi: cy cn ty, cy cn tu [9].

9

1.2.2. c im thc vt

Cy tho, cao 15-150cm, ton thn c mi thm, sng 1-2 nm. Thn mc

thng ng, nhn, c nhiu rnh dc, phn nhnh nhiu [11], [13]. L hnh thun

hoc hnh trng ngc, di 7-18cm, rng 3,5-8cm, chia lm 3 thy hoc x 3, thy

cui c hnh thoi, kch thc 1,2-2,5 0,8-2,5cm, c rng ca hoc kha tai bo. L

pha trn c cung ngn, phin l hnh tam gic rng, x su 3 thy, thy cui c

hnh trng [32]. L gc c cung, b to rng, c nhiu sng, hnh tam gic

thun hoc dng 5 cnh c gc ct, x 3-5 thy hnh tam gic, u t, mp kha rng

to, khng lng. Cm hoa dng tn, rng 1,5-4cm, mc i din vi l, gm nhiu

tn di, gn khng u, cc tn u c cung di hn cc tn bn trong v c kch

thc 4-15mm [13], [32], tn kp mang 8-12 tn n [9], [13], mi tn hoa c 7-25

hoa, kch thc 6-9mm theo chiu ngang. Hoa pha ngoi c 3-8(-16) cnh hoa

mnh, kch thc 0,5-2,5cm [32]. Hoa nh mu trng hoc lc nht, trng c cnh

khum, bu nh [13]. Qu i dng trng, hi dt, nhn, c cnh li chy dc thn

[13], c 5 cnh, 2n=22 [9], kch thc 1,3-1,5 1-2mm. Cung qu di 1-1,5mm,

qu c kch thc 1,3-1,5 1-2mm [32].

Ma ra hoa v qu t thng 4 n thng 7 [32].

Hnh 1.4. Cy cn ty ( Apium graveolens L.) [11]

10

1.2.3. Phn b

Loi Apium graveolens L. c ngun gc b bin i Ty Dng v a

Trung Hi, c trng lu i cc nc phng Ty [4], thch hp v phn b

nhiu vng kh hu n i. Loi Apium graveolens L. c 3 th sau [13]:

- A.graveolens L. var. secalium Alef (cn ty cho l): trng nhiu Trung

Quc, Vit Nam v cc nc ng Nam khc.

- A.graveolens L. var. dulce (Miller) Pers (cn ty cho cung): trng nhiu

vng n i m ca chu u hay chu (ty Lin bang Nga, Ucraina .

- A.graveolens L. var. rapaceum (Miller) Gaudin (cn ty cho c): trng

chu u.

Nhn chung, tt c cc loi cn ty u a kh hu m mt, nhit trung bnh

t 15 n 210C ( Vit Nam v ng Nam [13]. V ma ng, cy chu c

nhit di 50C trong vi ngy, khng chu c nng nng. Qun th cn ty

mc hoang di chu u v mt vi ging cy trng Trung Quc c th chu

c hn. Cy ra hoa, qu nhiu; ti sinh t nhin ch yu t qu [5], [13].

Vit Nam, loi A. graveolens L. thng gi l cy cn ty. Cy mi di

nhp vo nc ta v c trng rng ri nhiu ni lm rau n.

1.2.4. Thnh phn ha hc ca ht cn ty

Cc nghin cu v cn ty cho thy c 89 cht c pht hin c trong ht

cn ty, ch yu thuc 3 nhm cht flavonoid, tinh du v coumarin, ngoi ra cn

c glycoside v mt s nhm cht khc. C th:

- Flavonoid: 10 cht.

- Tinh du: 28 cht.

- Coumarin: 22 cht.

- Glycosid: 16 cht.

- Cc nhm cht khc: 13 cht.

1.2.4.1. Flavonoid

Cc flavonoid c trong ht cn ty gm 10 cht c khung cu trc c trnh

by hnh 1.5 v c tm tt bng 1.1.

11

Hnh 1.5. Khung cu trc ca cc hp cht flavonoid c trong ht cn ty

Bng 1.1. Cc hp cht flavonoid c trong ht cn ty

ST

T Tn cht R1 R2 R3

Tc dng

sinh hc

(nu c)

TL

TK

1 Apigenin -H -H -H

Gin mch,

chng kt

tp tiu cu,

c ch men

chuyn v

chn knh

canxi

[36]

[37]

2

Apigenin-7-O-

[2-O- 5-O-

feruloyl)--d-

apiofuranosyl]-

-d-

glucopyranosid

-H

-H [46]

3

Apiin

(Apigenin-7-O-

apiosyl

glucosid)

-H

-H Chng vim [23]

12

4

Chrysoerinol-7-

O-apiosyl

glucosid

-OCH3

-H [23]

5 Chrysoerinol-7-

O-glucosid -OCH3

-H [46]

6

Chrysoeriol-7-

O-[2-O- 5-O-

feruloyl)--d-

apiofuranosyl]-

-d-

glucopyranosid

-OCH3

-H [46]

7 Luteolin -OH -H -H [25]

8

Luteolin-3-

methylether-7-

apiosyl glycosid

-OH

-OCH3 [13]

9 Luteolin-7-O-

apiosyl glucosid -OH

-H [23]

10 Luteolin-7-O-

glucosid -OH

-H [23]

Cc flavonoid c trong cn ty ch yu c khung flavon, l dn xut ca

apigenin hoc luteolin. Apigenin v dn cht l nhm cht c tc dng sinh hc

mnh vi nhiu tc dng quan trng nh tc dng chng vim, chng ngng tp

tiu cu, chng ung th.

13

1.2.4.2. Tinh du

Ht cn ty cha 2 - 3% tinh du [13]. Kt qu nh lng tinh du trong ht

cn ty mt s a phng cho thy ht cn ty H Ni cha 0,08% tinh du [1],

ht cn ty Nam nh cha 0,88% tinh du [1].

Cho n nay theo cc ti liu cng b, bng phng php sc k kh v khi

ph pht hin ra c 28 thnh phn trong tinh du ht cn ty, trong :

- 7 hp cht phthalid, chim khong 37,4 - 42,3%.

- 11 hp cht monoterpenoid.

- 6 hp cht sesquiterpenoid.

- 4 hp cht khc.

Tin hnh phn tch c th cc cht trong nhm hp cht trn thy: tinh du c

trong ht cn ty ch yu l limonen v -selinen, sedanolid, sedanenolid,

3nBTrong , 3nB v sedanoid l nhng hp cht c nhiu tc dng sinh hc:

3nB c tc dng h huyt p, chng vim, bo v gan, chng ung th v lm gim

c tnh ca acrylamid. Sedanoid c tc dng gii c gan, chng lot d dy - t

trng, chng oxy ha.

1.2.4.3. Coumarin

Cc nghin cu v cn ty cho thy c 22 cht thuc nhm coumarin, trong

11 cht thuc nhm fruocoumarin, 6 cht thuc nhm coumarin glycoside v 5 cht

thuc nhm coumarin n gin.

1.2.5. Tc dng sinh hc ca ht cn ty

1.2.5.1. Tc dng chng vim

Nhiu nghin cu gn y trn ng vt thc nghim chng minh tc

dng chng vim tt ca dch chit cn ty. Nghin cu ca Mencherini T v cng

s chng minh cc flavonoid trong l cn ty c tc dng chng vim trn thc

nghim [25]. Mt nghin cu khc c cng b cho thy tc dng chng vim tt

ca luteolin - mt flavonoid c trong cn ty trn thc nghim [49]. Cc nghin cu

thc hin trn m hnh gy vim lot d dy cng cho thy cn ty cho tc dng

chng vim tt.

14

1.2.5.2. Tc dng h acid uric mu

Dch chit ether du ha v dch chit methanol t ht cn ty c tc dng h acid

uric mu. Nghin cu c thc hin trn chut cng trng mc liu 500 mg/kg.

Kt qu cho thy nng acid uric ca l ung dch chit ether du ha v l ung

dch chit methanol t ht cn ty gim ln lt 41% v 38% so vi l chng ti

thi im 3 gi sau khi gy tng cp acid uric. Ti thi im 6 gi, nng acid

uric ca l ung dch chit ether du ha v l ung dch chit methanol t ht cn

ty gim ln lt 44% v 31% so vi l chng [14].

1.2.5.3. Tc dng chng oxy ha

Nm 2006, mt nghin cu chng minh rng dch chit r v l cn ty c

tc dng dn dp cc gc t do v lm gim cng lipoperoxy ha trong

liposome. Dch chit n-butanol ca cn ty cho hiu qu chng oxy ha cao nht

[11]. Mt nghin cu cng b nm 2011 cho thy hp cht phenolic trong ht cn

ty c tc dng kh gc t do, chng oxy ha mnh. ng lu , trong cn ty c 2

flavonoid l apigenin v kaempferol c chng minh c tc dng c ch enzym

xanthin oxidase [45].

1.2.5.4. Tc dng chng ung th

Nm polyacetylen trong dch chit cn ty c chng minh c tc dng gy

c t bo gm: falcarinol, falcarindiol, panaxydiol, 8-O-methylfalcarindiol. Kh

nng ny c nh gi bng xt nghim V-PI annexin. Falcarinol c chng

minh l hot cht c c tnh r rt trn dng t bo bch cu lympho trong bnh u

lympho cp. Dch chit cn ty cng chng t tc dng dit t bo ung th gan [35].

1.2.5.5. Tc dng khng khun

Dch chit cn ty c tc dng khng khun mnh. Theo nghin cu ca

Magrinya v cng s , dch chit cn ty c tc dng khng Staphylococus carnosus

[24]. Theo mt nghin cu khc, carvacrol v cinnamaldehyd t cn ty lm bt

hot vi khun Salmonella enteric khng khng sinh [30]. Dch chit ht cn ty c

tc dng chng vi khun Helicobacter pylori, mt trong nhng nguyn nhn gy

lot d dy - t trng [47].

15

1.2.5.6. Tc dng h huyt p

Nghin cu in vitro cho thy, dch chit methanol ht cn ty c tc dng c

ch men chuyn angiotensin. Tuy nhin, tc dng ny cn km xa so vi lisinopril

[41]. Tc dng h huyt p ca ht cn ty l do hp cht 3-n-butylphtalid (3nB) c

trong tinh du ht cn ty. C ch ca tc dng ny c th do s phong ta knh

Ca2+

thng qua in p hot ng hoc th th trn knh Ca2+ [40].

1.2.6. Cng dng ca ht cn ty

Ht cn ty c s dng trong cc bi thuc y hc c truyn cha thng

phong [8], vim bng quang, gip kh trng bng quang v ng dn nc tiu [8],

[13]; kch thch tiu ha, gy trung tin, b sung vitamin C v bi b c th [13];

cha cc bnh suyn, hen ph qun [8]; cha cc bnh v khp nh thp khp, vim

khp, au khp [13]. Ngoi ra, trong y hc c truyn, tinh du cn ty c s dng

trong bnh ph thng, au bng quang, cc trng hp co tht c trn.

16

CHNG 2: I TNG V PHNG PHP NGHIN CU

2.1. NGUYN LIU NGHIN CU

2.1.1. Dc liu nghin cu

- Cy cn ty c hoa v ht c thu hi ti x Hi Nam - Hi Hu - Nam

nh vo thng 11/2012.

- X l mu:

Mu cy c hoa c p tiu bn, lu tr ti phng tiu bn - b mn Thc

vt - trng i hc Dc H Ni vi s hiu HNIP/17860/13.

Cn c vo c im hnh thi ca mu nghin cu, s dng kha phn loi

chi pium v i chiu vi bn m t loi theo ti liu [39], mu cn ty nghin

cu c xc nh tn khoa hc l Apium graveolens L., h Cn ( Apiaceae).

Ht cn ty c em phi kh, bo qun ni kh ro, thong mt.

2.1.2. Chun b dch chit dc liu

Ngm bt dc liu xay th vi dung mi ethanol 96% nhit phng

trong 72 gi. Lp li 3 ln.

Dch chit sau 3 ln ngm c ct thu hi dung mi di p sut gim. Loi

b cht bo bng cch lc vi ether du ha. Sn phm thu c c sy trong t

sy chn khng n khi tr thnh cao kh.

Quy trnh chit cao ton phn ht cn ty c thc hin nh hnh 2.1.

17

Hnh 2.1. Quy trnh chit xut cao ton phn ht cn ty

2.2. PHNG TIN NGHIN CU

2.2.1. ng vt th nghim

- Chut nht trng chng Swiss, ging c, cn nng t 18-22g, khe mnh

do Vin V sinh dch t trung ng cung cp.

- Chut cng trng chng Wistar, ging c, 8 tun tui, cn nng t 130-

150g, khe mnh do Hc vin Qun y cung cp.

ng vt c nui n nh vi iu kin phng th nghim t nht 5 ngy

trc khi thc hin nghin cu, c nui dng bng thc n tiu chun do Vin

V sinh dch t trung ng cung cp, ung nc t do.

2.2.2. Ha cht, thuc th

- Allopurinol (Sigma Aldrich).

18

- cid uric, 99%, dng tinh th, dng cho nghin cu v pht trin (Sigma

Aldrich).

- B ha cht nh lng acid uric huyt thanh gm ha cht phn ng v

acid uric chun (Teco Diagnostics).

- Carrageenan (Sigma Aldrich).

- Indomethacin, tiu chun phn tch (Vin Kim nghim thuc trung ng -

B Y t).

- Ha cht pha mu dc liu: dimethyl sulfoxid (Merck).

- Kali oxonat, 97%, dng cho nghin cu v pht trin (Sigma Aldrich).

- Xanthin, 99% Sigma ldrich .

- Xanthine oxidase t sa b (0,8 U/mg protein, 9 mg protein/ml, Sigma

Aldrich).

- Cc dung mi cho chit xut: ethanol, ether du ha t tiu chun cng

nghip.

- Cc dung mi, h m dng cho phn ng enzym t tiu chun phn tch.

- Cc ha cht, dung mi khc t tiu chun dc dng.

2.2.3. Thit b

- My sinh ha TC 84 plus (Teco Diagnostics).

- My o ph chn chut LE 7500.

- My ly tm HERMLE Z300.

- My ly tm lnh Sigma 3-18K (Sartorius).

- Thit b nghin ng th.

- H thng ELISA gm my c khay vi tinh th Biotek, Hoa K v my

lc khay (Awareness, Hoa K).

- My o pH EUTECH).

- My ct quay Buchi Rotavapor R210.

- Cn phn tch AY 220 (SHIMADZU).

- Cn k thut Presica-BJ610C, TE412 (Sartorius).

19

- T m iu nhit Memmert.

- Cc dng c s dng ly mu v xt nghim: micropipet v u cn tng

ng, dng c thy tinh, bm kim tim cc loi, mao qun, ng nghim cc loi.

2.3. NI DUNG NGHIN CU

t c cc mc tiu nghin cu t ra, ti c thit k vi cc

ni dung tin hnh nh sau:

- nh gi tc dng h acid uric ca cy cn ty trn thc nghim.

- nh gi tc dng c ch xanthin oxidase in vitro v in vivo.

- nh gi tc dng chng vim ca cy cn ty trn thc nghim.

Thit k ni dung nghin cu c m t hnh 2.2.

Hnh 2.2. Thit k nghin cu

20

2.4. PHNG PHP NGHIN CU

2.4.1. Phng php nh gi tc dng h acid uric huyt thanh trn m hnh gy

tng acid uric cp thc nghim.

Thit k th nghim

nh gi tc dng h acid uric huyt thanh trn m hnh gy tng acid uric cp

bng kali oxonat [6], [48], [26].

Chut nht trng, c, trng thnh c chia ngu nhin thnh cc l:

- L chng: ung dung mi.

- L i chiu: ung thuc i chiu allopurinol.

- Cc l th: ung cao ton phn ht cn ty cc mc liu th nghim

Quy trnh th nghim c m t hnh 2.3

4 ngy Ngy th nm

Tim kali oxonat

Nhn n Ung thuc Ly mu

Ung thuc

hng ngy 30 pht 60 pht 120 pht

Hnh 2.3. Quy trnh th nghim nh gi tc dng h acid uric mu trn m

hnh gy tng cp bng kali oxonat

Thng s nh gi

- Nng acid uric trong huyt thanh chut th nghim.

- T l gim nng acid huyt thanh ca l th so vi l chng.

I (%)=

Trong :

Cc, Ct: nng acid uric huyt thanh ca l chng, l th.

I (%): t l gim acid uric huyt thanh ca th so vi chng trng.

21

2.4.2. Phng php nh gi tc dng c ch xanthin oxidase in vitro v in vivo

2.4.2.1. Phng php nh gi tc dng c ch xanthin oxidase in vitro

Nguyn tc

XO xc tc cho qu trnh oxy ha xanthin thnh acid uric:

Xanthin + O2 + H2O uric acid + H2O2

Hot XO t l vi lng acid uric nh lng c bng phng php o

quang bc sng 290nm.

Thit k th nghim

Th nghim c thc hin trn a 96 ging.

B tr trong mt a gm: mu chng v mu th.

Thnh phn trong mu chng v mu th c th hin bng 2.1.

Bng 2.1. Thnh phn mu chng v mu th

Mu chng Mu th

Dung dch th 0 50l

Dung dch m

phosphate pH 7,5 85 l 35l

Xanthin 150M 60l 60l

XO 0,01U/ml 30l 30l

HCl 1M 25l 25l

Tng th tch 200l 200l

XO

22

Quy trnh th nghim c m t hnh 2.4

Hnh 2.4. Quy trnh th nghim nh gi tc dng c ch XO in vitro

Thng s nh gi

- OD:

ODchng = ODchng - ODtrng chng.

ODth = ODth - ODtrng th

- I (%) c ch tnh theo cng thc:

- IC50 c tnh: l nng l thuyt m ti c ch 50% hot xanthin

oxidase.

2.4.2.2. Phng php nh gi tc dng c ch xanthin oxidase in vivo

Thit k th nghim

p dng m hnh nh gi nh hng ln hot XO gan chut thc nghim [6].

Chut nht trng, c, trng thnh c chia ngu nhin vo cc l:

- L chng: ung dung mi.

- L i chiu: ung thuc i chiu allopurinol.

- Cc l th: ung cao ton phn ht cn ty cc mc liu th nghim.

23

Phng php chit enzym

Kt thc thc nghim, git chut bng cch ko dn t sng c, ly ngay

gan tng chut. Cn ngay 1g gan, ct thnh cc mnh nh, cho vo ng nghin ng

th lnh v c sn 1ml dung dch m phosphate lnh (pH = 7,5). Nghin gan

trong iu kin lnh n khi thu c dch ng th, sau thm dn m lnh

thu c dch ng th vi t l gan : m tng ng l 1:10. Dch nghin thu c

em ly tm lnh 00C vi tc 3000 vng trong 10 pht. Ly phn dch ni tip

tc em ly tm lnh 00C vi tc 10000 vng trong 60 pht. Phn dch trong thu

c sau khi ly tm dng xc nh hot enzym. Ton b cc bc ca th

nghim c thc hin trong iu kin lnh (mi trng nc ang tan .

Phn ng enzym

Phn ng c thc hin trong ng nghim gm 2,90ml m phosphate

(0,05M, pH 7,5); 2,50ml dung dch m c cht gm xanthin 0,12mM, EDTA

0,192mM, kali oxonat 2,2mM. Trc khi phn ng, dch enzym v c cht c

370C trong 15 pht. Thi gian phn ng c tnh t lc thm 0,1ml dch enzym

vo dung dch c cht. Tip tc trong 30 pht. Ht 200l dch sau phn ng ra a

Costar 96 ging tin hnh o quang bc sng 290nm.

Song song tin hnh vi mu trng: 5,40ml m phosphate pH 7,5 + 0,1ml

enzym.

Thng s nh gi

nh hng ca cao ton phn ht cn ty ln hot XO trn gan chut thc

nghim s c nh gi thng qua s bin thin mt quang OD ca acid uric

sn phm c to ra trong phn ng gia c cht xanthin vi enzym XO c

chit ra t gan chut ca l th so vi l chng.

- Mt quang o c ca tng mu:

ODmu = ODth - ODtrng ca th

Trong :

ODth: mt quang ca mu th.

ODtrng: mt quang ca mu trng ca th.

24

- T l gim mt quang ca l th so vi l chng trng.

I(%) =

Trong :

ODc, ODt: mt quang ca cc mu l chng, l th.

I (%):t l gim mt quang acid uric ca l th so vi l chng.

2.4.3. Phng php nh gi tc dng chng vim thc nghim

2.4.3.1. nh gi tc dng chng vim cp trn m hnh gy ph bn chn chut

bng carrageenan

Thit k th nghim

S dng m hnh gy ph bng carrageenan theo Winter [42].

Chut cng trng c c chia ngu nhin thnh cc l:

- L chng: ung dung mi.

- L i chiu: ung indomethacin.

- Cc l th: ung cao ton phn ht cn ty cc mc liu th nghim.

Quy trnh th nghim c m t hnh 2.5.

4 ngy Ngy th nm

- Ung thuc

- Gy vim

Nhn n

Ung thuc

hng ngy 1,5 gi

V0 V1 V3 V5 V7

o th tch bn chn gy vim

Hnh 2.5. Quy trnh nh gi tc dng chng vim cp trn m hnh gy ph

bn chn chut bng carrageenan

25

Thng s nh gi

- Th tch bn chn sau phi ca tng chut.

- Mc ph bn chn sau phi ca tng chut c tnh theo cng thc:

V (%) = [(Vt-V0)/Vo] 100

V: Mc ph bn chn chut ti thi im t gi sau khi gy vim.

V0, Vt: Th tch bn chn chut trc v sau khi gy vim t gi.

- % c ch ph ca cc l th so vi l chng c tnh theo cng thc:

I (%) =

I (%): Phn trm c ch ph ca l th so vi l chng.

Vc, Vt: Mc ph chn chut trung bnh l chng v l th.

2.4.3.2.Phng php nh gi tc dng chng vim trn m hnh gy vim mng

hot dch khp gi

S dng m hnh gy vim mng hot dch khp gi bng tinh th natri urat ca

McCarty v Faires [42].

Chun b hn dch tinh th natri urat

Ha tan 0,4g 0,01 mol natri hydroxid trong 400ml nc ct. Thm 1,68g (0,01

mol acid uric. qua m nhit phng. Lc ht chn khng thu c tinh th,

ra 3 ln bng nc mui sinh l lnh. To li hn dch bng nc mui sinh l v

tit trng trong ni hp. Hn dch natri urat thu c cui cng c nng 48

mg/ml.

Thit k th nghim

Chut cng trng c c chia ngu nhin thnh cc l:

- L chng: ung dung mi.

- L i chiu: ung thuc i chiu indomethacin.

- Cc l th: ung cao ton phn ht cn ty cc mc liu th nghim.

. Thng s nh gi

S dng bng thang im nh gi mc vim da trn triu chng nh

bng 2.2 sau:

26

Bng 2.2. Thang im nh gi mc vim da trn triu chng

Cc quan st vin c hun luyn cch quan st v thng nht cch nh gi

trc khi lm thc nghim chnh thc. Kt qu cui cng c ghi nhn t quan st

ca 3 quan st vin c lp v p dng phng php cho im m.

2.4.4. Phng php x l s liu

Kt qu c biu din di dng M SE (M: gi tr trung bnh ca tng l,

SE: sai s chun). So snh gi tr trung bnh gia cc l bng one-way ANOVA,

dng hu kim so snh gi tr trung bnh ca cc l th so vi l chng.

Vi cc s liu khng thuc phn phi chun, kt qu c trnh by di

dng trung v v khong dao ng (gi tr cc tiu - gi tr cc i). Dng thut ton

Kruskal Wallis so snh gia cc l, Mann-Whitney test so snh kt qu gia

l th v l chng.

IC50 v khong tin cy 95% c xc nh bng phng php hi quy probit.

Phn tch thng k bng phn mm SPSS 20.0, s khc bit gia cc l c

coi l c ngha khi p < 0,05.

Triu chng M t im

Khng th hin r triu

chng

Chut khng c hoc khng th hin r biu

hin di chuyn bt thng 1

i khp khing Chut i tp tnh trong ton b thi gian di

chuyn, chn c khp b au vn tham gia

vo qu trnh di chuyn

2

i bng 3 chn khng

lin tc

Chut i tp tnh nhng thnh thong chn c

khp b au khng th tham gia vo qu trnh

di chuyn, phi ko l hoc co chn

3

i hon ton bng 3

chn

Chn c khp b au khng th tham gia vo

qu trnh di chuyn, khng th c ng khp

b vim, phi ko l hoc co chn

4

27

CHNG 3: THC NGHIM V KT QU

3.1. KT QU NH GI TC DNG H ACID URIC CA CAO TON

PHN HT CN TY

Chut nht c nui n nh trong phng th nghim v c chia ngu

nhin thnh 4 l:

- L chng: ung dung mi pha thuc Na-CMC 0,5%.

- L i chiu: ung allopurinol 10 mg/kg.

- L cao ton phn 1: ung cao ton phn ht cn ty liu 250 mg/kg pha

trong Na-CMC 0,5%.

- L cao ton phn 2: ung cao ton phn ht cn ty liu 500 mg/kg pha

trong Na-CMC 0,5%.

Chut c ung thuc vi cng th tch 0,1ml/10g vo mt gi nht nh

hng ngy trong vng 5 ngy trc khi lm th nghim. Trc khi dng dung mi

hoc thuc 1,5 gi, chut khng c n nhng c ung nc bnh thng. Ngy

th 5, chut cc l c tim mng bng kali oxonat vi liu 500 mg/kg 1 gi

trc khi ung thuc ln cui. Sau khi ung thuc 2 gi, ly mu t xoang hc mt

ca chut, lng t nhin nhit phng 1 gi trc khi em ly tm vi tc

3500 vng trong 10 pht ly huyt thanh. Xc nh nng acid uric trong huyt

thanh chut th nghim.

Kt qu c biu din bng 3.1 v hnh 3.1.

28

Bng 3.1. Kt qu nh gi tc dng h acid uric ca cao ton phn ht cn

ty trn m hnh gy tng acid uric cp

L Liu dng

( mg/kg) n

Nng acid uric

(mol/l)

T l gim so vi

chng bnh (%)

Chng bnh - 15 259,01 22,87

Allopurinol 10 17 52,56 4,75** 79,7

Cao ton phn 1 250 17 105,41 10,41** 59,3

Cao ton phn 2 500 15 122,81 12,87** 52,5

**, p < 0,01 khi so snh vi l chng bnh

Hnh 3.1. Kt qu nh gi tc dng h acid uric ca cao ton phn ht cn ty

trn m hnh gy tng acid uric cp, ** p < 0,01 khi so snh vi l chng bnh

Nhn xt:

- Cao ton phn cn ty vi liu 250 mg/kg v 500 mg/kg c tc dng gim

nng acid uric huyt thanh chut nht trng th nghim trn m hnh gy

29

tng acid uric cp bng kali oxonat, t l gim so vi l chng bnh ln lt

l 59,3% v 52,5% (p < 0.01).

- Thuc i chiu allopurinol liu 10 mg/kg c tc dng h acid uric huyt

thanh r rt t l gim so vi chng bnh l 79,7% (p < 0,01).

- Khi so snh nng acid uric huyt thanh gia hai l th dng liu 250

mg/kg v 500 mg/kg, s khc bit khng c ngha thng k.

3.2. KT QU NH GI TC DNG C CH XO IN VITRO V IN

VIVO CA CAO TON PHN HT CN TY

3.2.1. Kt qu nh gi tc dng c ch XO in vitro ca cao ton phn ht cn

ty

Khi mt thuc lm gim nng acid huyt thanh, c ch tc dng thng

c d on theo 2 hng: lm gim tng hp acid uric (thng do c ch XO)

v/hoc lm tng thi tr acid uric. Trong , hng c ch XO thng c ngh

n nhiu hn. V vy, bc u, chng ti tin hnh nh gi tc dng c ch XO

in vitro ca cao ht cn ty.

Mu cao ht cn ty s c th tc dng c ch XO in vitro ti 3 nng 100

g/ml, 50 g/ml, 10 g/ml. Mi nng c tin hnh th ba ln, mi ln th ba

ging.

Kt qu nghin cu c trnh by bng 3.2.

Bng 3.2. Kt qu nh gi tc dng c ch XO in vitro ca cao ton phn

ht cn ty

Nng OD % c ch I (%)

100 g/ml 0,072 0,013* 66,8

50 g/ml 0,099 0,013* 53,3

10g/ml 0,145 0,019* 31,3

IC50 c tnh ( g/ml) = 52,08 (36,19 67,56)

*, p < 0,05 khi so snh vi mu chng

30

Nhn xt:

- Cao ton phn ht cn ty nng 100 g/ml th hin kh nng c ch XO

in vitro vi t l c ch l 66,8%, OD = 0,072 0,013 p < 0,05).

- Cao ton phn ht cn ty nng 50 g/ml th hin kh nng c ch XO

in vitro vi t l c ch l 53,3%, OD = 0,099 0,013 (p < 0,05).

- Cao ton phn ht cn ty nng 10 g/ml th hin tc dng c ch XO in

vitro vi t l c ch l 31,3%, OD = 0,145 0,019 p < 0,05).

- Gi tr IC50 c tnh l 52,08 (36,19 67,56 g/ml.

T kt qu trn, chng ti tip tc nghin cu su hn v kh nng c ch XO

in vivo ca cao ton phn ht cn ty.

3.2.2. Kt qu nh gi tc dng c ch XO in vivo ca cao ton phn ht cn

ty

Chut nht trng c nui n nh trong phng th nghim v c chia

ngu nhin thnh 4 l:

- L chng: ung dung mi Na-CMC 0,5%.

- L i chiu: ung allopurinol liu 10 mg/kg.

- L cao ton phn 1: ung cao ton phn ht cn ty liu 250 mg/kg pha

trong Na-CMC 0,5%.

- L cao ton phn 2: ung cao ton phn ht cn ty liu 500 mg/kg pha

trong Na-CMC 0,5%.

Chut c ung dung mi pha thuc, thuc i chiu hoc ch phm th

vi cng th tch 0,1ml/10g vo mt gi nht nh trong vng 5 ngy trc khi lm

thc nghim. Trc khi dng dung mi hoc thuc 1,5 gi , chut khng c n

nhng c ung nc bnh thng.

nh hng ca cao ton phn cn ty ln hot xanthin oxidase trn gan

chut thc nghim s c nh gi thng qua s bin thin mt quang OD ca

acid uric sn phm c to ra trong phn ng gia c cht xanthin vi enzym

xanthin oxidase c chit ra t gan chut ca l th so vi l chng.

31

Kt qu c trnh by bng 3.3 v hnh 3.2.

Bng 3.3. Kt qu nh gi tc dng c ch XO in vivo ca cao ton phn ht

cn ty

L Liu dng

(mg/kg) n

Mt quang

acid uric

T l gim mt

quang OD so vi

chng (%)

Chng bnh - 10 0,457 0,014

Allopurinol 10 10 0,377 0,007** 17,8

Cao ton phn 1 250 9 0,402 0,015* 12,1

Cao ton phn 2 500 10 0,409 0,040* 10,5

*, p < 0,05 v **, p < 0,01 khi so snh vi l chng bnh

Hnh 3.2. Kt qu nh gi tc dng c ch XO in vivo ca cao ton phn

ht cn ty, * p < 0,05, ** p < 0,01 khi so snh vi l chng bnh

32

Nhn xt:

- Cao ton phn ht cn ty liu 250 mg/kg v 500 mg/kg c tc dng lm

gim mt quang ca acid uric sn phm ca phn ng gia c cht

xanthin vi enzym xanthin oxidase c chit t gan chut thc nghim, t

l gim so vi l chng bnh ln lt l 12,1% v 10,5% (p < 0,05).

- Thuc i chiu allopurinol vi liu 10 mg/kg c tc dng lm gim mt

quang acid uric 17,8% (p < 0,01) so vi l chng bnh.

- Khi so snh mt quang acid uric gia 2 l th mc liu 250 mg/kg v

500 mg/kg, s khc bit khng c ngha thng k.

3.3. KT QU NH GI TC DNG CHNG VIM CA CAO TON

PHN HT CN TY

3.3.1. Kt qu nh gi tc dng chng vim trn m hnh gy ph bn chn

chut bng carrageenan

Chut cng trng c nui n nh trong phng th nghim v c chia

ngu nhin thnh 4 l:

- L chng: ung dung mi Na-CMC 0,5%.

- L i chiu: ung indomethacin vi liu 10 mg/kg.

- L cao ton phn 1: ung cao ton phn ht cn ty liu 250 mg/kg pha

trong Na-CMC 0,5%.

- L cao ton phn 2: ung cao ton phn ht cn ty liu 500 mg/kg pha

trong Na-CMC 0,5%.

Chut cc l c ung thuc tng ng vi cng th tch 1ml/100g chut

vo mt gi nht nh hng ngy trong vng 5 ngy trc khi lm thc nghim.

Trc khi dng thuc 1,5 gi chut khng c n nhng c ung nc bnh

thng. Ngy th 5, s dng my o ph bn chn chut LE 7500 o th tch

bn chn sau phi ca tng chut. Sau khi ung dung mi, thuc i chiu hoc ch

phm nghin cu ln cui, chut c tim 0,1ml carrageenan 1% trong nc mui

33

sinh l vo gan bn chn sau phi. o th tch bn chn sau phi ca tng chut vo

cc thi im 1, 3, 5, 7 gi sau khi gy vim.

Kt qu th hin trn bng 3.4 v hnh 3.3.

Bng 3.4. Kt qu nh gi tc dng chng vim trn m hnh gy ph

chn chut bng carrageenan

L Mc ph chn chut theo thi gian

1 3 5 7

Chng (n=9)

M SE

19,16 3,33

26,01 2,49

14,26 1,97

9,18 1,51

Indomethacin

10mg/kg

(n=8)

M SE (%)

I (%)

6,53 1,69**

65,9

13,09 3,17**

49,6

8,38 2,58

41,2

5,59 1,79

39,1

Cn ty 250

mg/kg (n=9)

M SE (%)

I (%)

11,94 2,80*

37,6

25,13 3,96

3,3

21,42 3,07

-

16,37 3,49

-

Cn ty 500

mg/kg (n=9)

M SE (%)

I (%)

7,54 1,54*

60,6

18,02 1,82

30,7

8,78 1,49

38,4

4,72 0,88

48,5

*, p < 0,05; **, p < 0,01 khi so snh vi l chng bnh

34

*, p < 0,05 v **, p < 0,01 khi so snh vi l chngbnh

Hnh 3.3. Kt qu nh gi tc dng chng vim trn m hnh gy ph chn

chut bng carrageenan

Nhn xt:

- Indomethacin liu 10 mg/kg l thuc c dng chng vim tt, tc dng xut

hin ngay t gi th nht (p < 0,01 v ko di n thi im 3 gi (p <

0,01), t l c ch ph bn chn chut so vi chng tng ng ln lt l

65,9% v 49,6%.

- Cao ton phn cn ty liu 250 mg/kg c tc dng c ch ph bn chn

chut ngay gi th nht (p < 0,05), t l c ch ph so vi chng tng

ng l 37,6%.

- Cao ton phn cn ty liu 500 mg/kg c tc dng c ch ph bn chn

chut ngay gi th nht (p

35

Chut cng trng c nui n nh trong phng th nghim v c chia

ngu nhin thnh 4 l:

- L chng: ung dung mi Na-CMC 0,5%.

- L chng bnh: ung indomethacin liu 10 mg/kg.

- L cao ton phn 1: ung cao ton phn ht cn ty liu 250 mg/kg pha

trong Na-CMC 0,5%.

- L cao ton phn 2: ung cao ton phn ht cn ty liu 500 mg/kg pha

trong Na-CMC 0,5%.

Th tch cho chut ung u l 0,1ml/10g chut. Trc khi dng thuc 1,5

gi, chut khng c n nhng c ung nc bnh thng. Ngy th 5, sau khi

ung dung mi, thuc i chiu v ch phm th 1 gi, chut c tim 0,05ml hn

dch natri urat (nng 48 mg/ml) vo khp gi sau phi. Quan st biu hin di

chuyn ca tng chut trn mt phng ti cc thi im 4, 5, 6 gi sau khi tim, ghi

li hnh nh bng my quay h tr nhn nh khi cn thit. S dng bng thang

im nh gi mc vim da trn triu chng.

Kt qu c trnh by bng 3.5.

Bng 3.5. Kt qu nh gi tc dng chng vim trn m hnh gy vim mng

hot dch khp gi bng tinh th natri urat

L

Liu

dng

(mg/kg)

n

im sau tim

4 gi 5 gi 6 gi

Chng - 9 4 (3-4) 4 (3-4) 3 (2-4)

Indomethacin 10 7 1 (1-1)** 1 (1-1)** 1 (1-1)**

Cao ton phn 1 250 10 2 (2-3)** 3 (2-4)* 3 (1-4)

Cao ton phn 2 500 10 2 (1-4)** 2 (1-4)** 2 (1-4)**

S liu c trnh by di dng trung v v khong ( cc tiu-cc i).

*, p < 0,05 v **, p < 0,01 khi so snh vi l chng, dng Mann-Whitney U test.

36

Nhn xt:

- Chut cng trng l chng trn m hnh ny c biu hin vim khp cp

kh nng do tinh th natri urat .

- Indomethacin liu 10 mg/kg c tc dng chng vim tt, im vim trong

sut thi gian quan st l 1 (1-1), gim r rt so vi l chng (p < 0,01).

- Cao ton phn cn ty liu 250 mg/kg th hin tc dng chng vim trn

m hnh ny. im vim ti thi im 4 gi sau tim l 2 (2-3), gim so

vi l chng (p < 0,01 . im vim ti thi im 5 gi sau tim l 3 (2-4),

gim so vi l chng (p < 0,05).

- Cao ton phn cn ty 500 mg/kg th hin tc dng chng vim tt trong

sut qu trnh quan st, im vim ti 3 thi im u l 2 (1-4), gim r

rt so vi l chng (p < 0,01).

37

CHNG 4: BN LUN

4.1. V TC DNG H ACID URIC CA CAO TON PHN HT CN

TY

Enzym uricase c gan ca hu ht cc loi ng vt c v (tr cc loi linh

trng), c vai tr chuyn ha acid uric thnh allantoin mt dng d tan hn v

o thi ra ngoi qua thn. Chnh v vy, cc loi ny, nng acid uric lun

mc bnh thng. c ch enzym uricase l mt phng php hiu qu v c

dng ph bin gy tng acid uric mu trn ng vt. Mt s cht c ch uricase

c dng gy tng acid uric mu trong cc nghin cu trn th th gii: kali

oxonat, acid oxonic v allantoxanamid. Trong nghin cu ny, chng ti s dng

kali oxonat lm cht c ch uricase gy tng acid uric vi mt s l do sau: Th

nht, kali oxonat c tc dng gy tng acid uric trong thi gian ngn, 2 gi sau khi

tim [22], [26], v 3 gi sau khi tim l thi im tng acid uric tng i n nh

trn chut nht trng [6], ph hp vi mc ch gy tng cp ca ti. Hn na,

mc tng acid uric trong mu ng vt l hp l, kt qu thu c cho php kt

lun cht th c hay khng c tc dng h acid uric huyt thanh trn c th sng.

Th hai, thi gian thc hin th nghim ngn (5 ngy do hn ch c cc sai s

do tc nhn khch quan gy ra, gim thiu chi ph cho nghin cu. Th ba, thng s

nh gi t, tng i n gin, hn ch ti a cc sai st trong qu trnh thc hin.

Mt nghin cu trn th gii v tc dng h acid uric ca cn ty c

cng b [14]. Nghin cu ny c thc hin trn chut cng trng vi mc liu

500 mg/kg. Dc liu c s dng l ht v l cn ty, chit xut bng dung mi

methanol v ether du ha. Da vo cng b ny, nghin cu ca chng ti tin

hnh trn chut nht trng, ti ba mc liu 250 mg/kg v 500 mg/kg v 1000 mg/kg.

Dc liu c s dng l ht cn ty v c chit xut bng phng php ngm

lnh vi dung mi ethanol 960, sau dch chit c loi b cht bo bng cch

lc vi ether du ha. Ethanol v methanol c kh nng ha tan cc cht mang tnh

chn lc, tuy nhin, methanol li c hn ethanol rt nhiu nn ti ca chng ti

38

la chn ethanol 960 lm dung mi chit xut cao ton phn. C ba mc liu nghin

cu u th hin kh tc dng h acid uric (kt qu ca mc liu 1000 mg/kg khng

c trnh by trong kha lun ny). Kt qu nghin cu cho thy, cao ton phn

ht cn ty vi hai mc liu 250 mg/kg v 500 mg/kg u c kh nng lm gim

nng acid uric huyt thanh chut nht trng thc nghim, t l gim so vi l

chng bnh ln lt l 59,3% v 52,5%. Kt qu ny tng ng vi nghin cu

c cng b trc [14] . Khi so snh nng acid huyt thanh gia 2 l th

dng liu 250 mg/kg v 500 mg/kg , s khc bit khng c ngha thng k. L gii

v tc dng h acid uric ca ht cn ty, chng ti hng n gi thuyt trong thnh

phn ca ht cn ty c nhng cht c kh nng c ch xanthin oxidase. iu ny s

c chng ti bn lun r hn phn sau. Ngoi ra, trong tinh du ht cn ty c

cha cc cht sedanolid, senkiunolide-N, senkiunolide-J, l nhng cht c kh nng

chng oxy ha [27], trong khi qu trnh chuyn ha acid uric gm mt lot cc

phn ng oxi ha xc tc enzym v gii phng ra H2O2, do chng oxy ha c

th dn n c ch qu trnh to ra acid uric, t lm gim nng acid uric trong

mu. Tuy nhin y ch l gi thuyt ban u ca chng ti. Chng dng

allopurinol lm gim r rt nng acid uric huyt thanh so vi l chng, t l

gim 79,7%. Allopurinol l thuc c tc dng h acid uric kh mnh, c la

chn hng u h acid uric trn bnh nhn gt. C ch tc dng ca thuc

c chng minh l c ch r rt xanthin oxidase, lm gim nng acid uric trong

mu v nc tiu [2].

Chng ti tip tc tin hnh nghin cu c ch h acid uric cng nh cc tc

dng khc ca ht cn ty ti hai mc liu 250 mg/kg v 500 mg/kg trn ng vt

thc nghim.

4.2. V KH NNG C CH XANTHIN OXIDASE CA CAO TON

PHN HT CN TY

4.2.1. V kh nng c ch XO in vitro

39

Chng ti p dng phng php ca nhm nghin cu Nguyn Th Thu

Hoi [10] nh gi tc dng c ch XO in vitro ca cao ton phn ht cn ty.

Phng php ny kh n gin, d thc hin v cho kt qu c chnh xc cao.

y, phn ng gia enzym xanthin oxidase v c cht xanthin s c khng ch v

mt thi gian, ng thi s dng acid HCl 1M lm cht dng phn ng. Qu trnh

tin hnh th nghim cn lu mt s im sau: Th nht, enzym xanthin oxidase

thng c bo qun nhit 2-40C. Nhng trong qu trnh tin hnh th

nghim nhit phng, rt c th enzym s c hot ha v sau gim hot

ng. V vy, trong nghin cu ny, chng ti phi lun bo m enzym c gi

trong mi trng lnh (nc ang tan cho ti khi ht vo tng a phn ng.

Th hai, c cht xanthin tan tt nhit cao, v vy, s dng phng php gia

nhit v khuy t tng tan ca xanthin ng thi gip rt ngn thi gian ha

tan. Xanthin s c pha mi trong cc ngy th nghim, khng s dng li xanthin

gia cc ngy.

Sau khi p dng phng php nh gi tc dng c ch in vitro ca cao

ton phn ht cn ty, kt qu bng 3.2 cho thy, c ba nng 100 g/ml, 50

g/ml v 10 g/ml u th hin tc dng c ch XO. T l c ch ln lt l 66,8%,

53,3% v 31,3%. Kt qu ny ca chng ti c s thng nht vi cc nghin cu

trn th gii cng b v tc dng c ch XO in vitro ca ht cn ty [45], [44].

IC50 c tnh ca cao ton phn ht cn ty l 52,08 (36,19-67,56) g/ml. Trong khi

, dc liu c IC50 nh hn 100 g/ml c coi l dc liu c tim nng, iu

ny gi cc nghin cu su hn v tc dng ca ht cn ty trong iu tr bnh

gt. Chng ti tip tc tin hnh nh gi tc dng c ch XO in vitro ca cao ton

phn ht cn ty.

4.2.2. V kh nng c ch XO in vivo

Trong phng php ca chng ti, nh hng ca cao ton phn ht cn ty

ln hot enzym xanthin oxidase s c nh gi gin tip thng qua s bin

thin mt quang acid uric gia cc l chut thc nghim. C s ca phng php

40

l da trn k thut o quang xc nh mt quang ca acid uric c to ra sau

phn ng. loi b nh hng ca mu sc gan ln mt quang o c, vi

mi mu th, chng ti tin hnh song song vi mu trng ca th trong cng

iu kin. Ngoi ra, khi s dng enzym trong dch nghin ng th t gan chut, rt

c th trong dch nghin c c enzym uricase - c tc dng chuyn acid uric thnh

allantoin khng hp th UV ti bc sng 290nm, iu ny s nh hng n kt

qu mt quang o c. Chnh v vy, loi b tc dng ca enzym ny, chng

ti thm cht c ch uricase (kali oxonat) vo dung dch c cht phn ng. Theo

nghin cu ca Hashimoto, khi cho kali oxonat vo hn hp nh lng vi nng

cui cng t 0,1mM n 1mM s khng lm nh hng n hot xanthin

oxidase ng thi c tc dng c ch tt enzym uricase [18]. V vy trong th

nghim ny, chng ti b sung thm kali oxonat 1mM. Trong nghin cu in

vitro, chng ti s dng acid HCl 1M lm cht dng phn ng. Nghin cu ny s

dng enzym tinh khit, do , HCl khng lm nh hng n mt quang. Tuy

nhin, trong nghin cu nh gi kh nng c ch XO in vivo, chng ti nhn thy

c s khc bit rt ln v mt quang o c khi cho v khng cho HCl. C th,

mu dng HCl lm cht dng phn ng c mt quang thp hn rt nhiu so vi

mu khng dng HCl. iu ny c th do HCl phn ng vi cht c mt trong

dch nghin ng th gan chut, to ra sn phm nh hng n kh nng hp th

quang. V vy chng ti khng s dng HCl 1M m ch khng ch thi gian

phn ng. a UV 96 ging Costar 3635 c chng ti s dng o quang ti

bc sng 290nm thay cho vic s dng cuvet. a UV 96 ging c th o quang

c nhiu mu cng lc, s lng mu o trong th nghim li kh nhiu, chnh v

vy vic thay th ny gip rt ngn ng k thi gian tin hnh th nghim, cng

nh khng ch c sai s v thi gian.

Kt qu t bng 3.3 cho thy cao ton phn cn ty mc liu 250 mg/kg v

500 mg/kg lm gim mt quang acid uric so vi l chng, t l gim tng ng

ln lt l 12,1% v 10,5%. Khi so snh mt quang gia 2 l th, s khc bit

khng c ngha thng k. Kt qu ny c th c l gii l do trong dch chit

41

ethanol ca cn ty c cha cc flavonoid c chng minh c kh nng c ch

xanthin oxidase [44], [45]. Luteolin l mt trong nhng flavonoid chnh c trong

dch chit ht cn ty, cht ny c chng minh l c tc dng c ch enzym

xanthin oxidase [44]. Luteolin s gn vo trung tm hot ng ca xanthin oxidase

v thay i cu trc th cp ca enzym ny. Ngoi ra hai flavonoid apigenin v

kaempferol cng c chng minh c tc dng c ch xanthin oxidase [45]. Tc

dng ny lin quan cht ch vi tc dng h acid uric. Vi kh nng c ch xanthin

oxidase, dch chit ethanol cn ty c kh nng lm gim tng hp acid uric, gp

phn quan trng vo tc dng h acid uric mu. Chng dng c s dng trong

nghin cu l allopurinol. Kt qu t bng 3.3 ch ra allopurinol c tc dng lm

gim mt quang acid uric so vi l chng, t l gim l 17,8%. Allopurinol l

mt thuc c chng minh c ch r rng l c ch xanthin oxidase. T nhng

kt qu trn y, nghin cu m ra trin vng pht trin, ng dng cn ty trong

iu tr gt, mt cn bnh c tc gia tng ngy cng nhanh chng trn th gii

ni chung v Vit Nam ni ring.

4.3. V TC DNG CHNG VIM CA CAO TON PHN HT CN

TY

4.3.1. V tc dng chng vim ca cao ton phn ht cn ty trn m hnh gy

ph bn chn chut bng carrgeenan

M hnh gy ph bn chn chut bng carrageenan c Winter khi

xng t nhng nm 60 ca th k trc [42] nhng cho n nay, y vn l m

hnh c s dng ph bin khi nghin cu v tc dng chng vim ca thuc trn

ng vt thc nghim. Khi gy ph bng carrageenan, phn ng vim gm hai giai

on khc nhau. Giai on 1 (0-2,5 gi sau khi tim carrageenan , c th gii phng

ra mt lot cc cht trung gian gy vim nh histamin, serotonin, kinin gy ph hy

xung quanh cc m. Giai on 2 (3-6 gi sau khi tim), i thc bo gii phng ra

bradykinin, leukotrien v prostagrandin. Giai on ny ko di do s gii phng ra

prostagrandin [43]. M hnh d p dng v kh nhy nh gi bc u v kh

42

nng chng vim ca mt thuc. Kt qu bc u ny s cho php kt lun s b

v tc dng v gi hng nghin cu trn cc m hnh tip theo.

Kt qu t bng 3.4 cho thy, chng dng indomethacin liu 10 mg/kg c

tc dng chng vim tt, tc dng xut hin ngay t gi th nht v ko di n gi

th 3. Dch chit ethanol cn ty mc liu 250 mg/kg v 500 mg/kg c tc dng c

ch ph bn chn chut ngay ti thi im u tin 1 gi. Kt qu ny ph hp vi

vic s dng cn ty trong bi thuc cha bnh thng phong [8] hay iu tr cc

bnh v khp [13].

4.3.2. V tc dng chng vim ca cao ton phn ht cn ty trn m hnh gy

vim mng hot dch khp bng tinh th natri urat

Sau khi xc nh c dch chit ethanol cn ty c tc dng chng vim cp

kh tt trn thc nghim, chng ti tip tc th nghim trn m hnh gy vim c

hiu hn i vi bnh gt. M hnh gy vim mng hot dch khp gi bng tinh

th natri urat c xy dng t nm 1962 [42]. Tc gi s dng phng php nh

gi mc vim theo thang im v y cng l phng php ti p dng.

Gn y, nghin cu theo m hnh ny c h tr bng cc thit b o gip cho

kt qu chnh xc hn [28]. M hnh gy vim mng hot dch khp gi bng tinh

th natri urat c c im gn vi c ch vim ca bnh gt. So vi m hnh gy

ph bn chn chut bng carrageenan, vic gy vim bng cch tim tinh th natri

urat vo khp gi ca chut s to ra c triu chng ging vi cn gt cp in

hnh. Do , m hnh ny rt ph hp nh gi tc dng chng vim ca thuc

iu tr gt.

Trong m hnh gy vim mng hot dch khp gi, sau khi tim hn dch

natri urat vo khp gi 4 n 6 gi, chut l chng c triu chng vim kh nng.

Hu ht phi di chuyn hon ton bng 3 chn, mt s di chuyn bng 3 chn khng

lin tc. Bng 3.5 cho thy, indomethacin liu 10 mg/kg v cao ton phn cn ty

liu 250 mg/kg v 500 mg/kg u c tc dng chng vim, trong indomethacin

th hin tc dng chng vim kh r rt. iu ny d l gii v indomethacin l mt

thuc chng vim mnh, c bit c ch nh trong vim cp do gt. Khi so snh

43

tc dng chng vim ca hai l th liu 250 mg/kg v 500 mg/kg, s khc bit

khng c ngha thng k. c bit, khi so snh vi indomethacin 10 mg/kg, tc

dng chng vim ca cao cn ty 250 mg/kg khng c s khc bit c ngha

thng k. Gi thuyt cho tc dng ny mt ln na c chng ti ngh n cc

flavonoid c mt trong dch chit ny. C rt nhiu flavonoid c chng minh l

c tc dng chng vim. Apiin l mt flavonoid ca ht cn ty c chng

minh c tc dng chng vim in vitro [25]. Theo nghin cu ny, c ch chng

vim ca luteolin lin quan n kh nng c ch enzym nitric oxide synthase, t

lm gim sn xut nitric oxide l mt cht trung gian gy vim. Luteolin c phn

lp t cn ty cng c chng minh c tc dng c ch chn lc cyclooxygenase-

2, t lm gim s xm nhp ca bch cu vo t chc vim, ng thi lm gim

sn xut cht trung gian gy vim 6-keto-prostagrandin F1 alpha [49]. Tc dng

chng vim ny ca luteolin th hin c hai m hnh gy vim cp v mn tnh

chut [49]. Bn cnh , hp cht flavonoid trong ht cn ty khng nh c

tc dng chng oxy ha nh chng ti trnh by mc 4.1.2. V vy, c th ngh

n kh nng thu dn cc gc t do gii phng trong qu trnh vim do i thc

bo, lm cn tr qu trnh peroxide ha lipid mng t bo, gy bt li cho qu trnh

gii phng acid arachidonic t phospholipid mng t bo, gim tng hp cc cht

trung gian ha hc gy vim. Tuy nhin, y ch l gi thuyt ban u ca chng

ti.

44

KT LUN V XUT

I. Kt lun

1. Tc dng h acid uric ca cn ty

Cao ton phn ht cn ty mc liu 250 mg/kg/ngy v 500 mg/kg/ngy, ung

lin tc trong 5 ngy c tc dng lm gim nng acid uric huyt thanh chut nht

trng thc nghim trn m hnh gy tng cp bng kali oxonat, t l gim so vi l

chng ln lt l 59,3% v 52,5%. Khi so snh hai l th ny, s khc bit khng

c ngha thng k.

2. Tc dng c ch xanthin oxidase in vitro v in vivo ca cn ty

Cao ton phn ht cn ty cc nng 100 g/ml, 50 g/ml, 10 g/ml th

hin kh nng c ch XO in vitro. T l c ch so vi mu chng ln lt l

66,8%, 55,3% v 31,3%. IC50 c tnh ca cao ton phn ht cn ty l 52,08

(36,1967,56 g/ml.

Cao ton phn ht cn ty mc liu 250 mg/kg/ngy v 500 mg/kg/ngy,

ung lin tc trong 5 ngy th hin kh nng c ch hot xanthin oxidase

gan chut nht trng, t l c ch ln lt l 12,1% v 8,06%. Khi so snh hai

l th ny, s khc bit khng c ngha thng k.

3. V tc dng chng vim ca cy cn ty

Trn m hnh gy ph bn chn chut bng carrageenan:

Cao ton phn ht cn ty liu 250 mg/kg v 500 mg/kg c tc dng c ch

ph bn chn chut so vi l chng, t l c ch ln lt l 65,9% v 49,6% ti thi

im 1 gi sau khi gy vim.

Trn m hnh gy vim mng hot dch khp gi bng tinh th natri urat:

- Cao ton phn ht cn ty liu 250 mg/kg lm gim triu chng vim ti thi

im 4 gi v 5 gi sau khi gy vim. im trung bnh ca l ung dch chit cn

ty ti hai thi im ln lt l 2 (2-3) v 3 (2-4) so vi l chng l 4 (3-4) c hai

thi im. Tc dng chng vim ny tng ng vi indomethacin.

45

- Cao ton phn ht cn ty liu 500 mg/kg lm gim triu chng vim ti c

ba thi im 4 gi, 5 gi, 6 gi sau khi gy vim. im trung bnh ca l ung dch

chit cn ty ti ba thi im u l 2 (1-4) so vi l chng l 4 (3-4) ti thi im 4

gi, 5 gi v 3 (2-4) ti thi im 6 gi.

II. xut

Cc kt qu thu c trong ti ny cho thy, cao ton phn ht cn ty c

tc dng h acid uric trn m hnh gy tng acid uric cp trn thc nghim, c ch

tc dng ban u c xc nh l do c ch xanthin oxidase. Bn cnh , cao cn

ty cn th hin tc dng chng vim. Cc tc dng ny cho php bc u kt lun

cn ty c trin vng trong iu tr gt. c th khai thc v s dng cn ty c

hiu qu hn trong cng tc phng v iu tr bnh xng khp ni chung v bnh

gt ni ring, chng ti xin a ra mt s xut:

- Xc nh phn on/ nhm cht/ cht ng vai tr quan trng trong tc

dng sinh hc ca cao ton phn ht cn ty.

- nh gi c tnh cp v bn trng din ca cao ton phn ht cn ty

c th a vo nghin cu trn lm sng.

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