BY: LAURA RODRÍGUEZ VERÓNICA PÉREZ

INTRODUCTUCTION

•PROGESTERONE

Is a steroid hormone involved in the female menstrual cycle and pregnancy.

Other functions are to increase the excretion of sodium. progesterone has antidepressant effects in some cases, by intervening directly in some intracellular receptors in the brain . It has been shown to be effective in treating some behavioral disorders.

Exist some studied of progesterone in

relation with brain tumors , and stopping their growth. This is

because progesterone regulates the

production of tumor cells by blocking the interaction with the

intracellular receptor tumor

AQP

– 4

Main function is to participate in metabolic

balance regulation in brain.

Its expression is closely related to the occurrence

and development of cerebral edema.

Transmembrane transport of wáter.

13 kinds ofAQPs

CEREBRAL ISCHEMIA• CEREBRAL ISCHEMIA INTERRUPTION OF SUPPLY BLOOD TO THE BRAIN ,

OCCURS WHEN BLOOD VESSELS THAT COME BRAIN BECOME NARROWED OR CLOGGED WITH FATTY DEPOSITS CALLED PLAQUE ( CAN BE CAUSED BY THROMBOSIS OR EMBOLISM) . THUS INTERRUPTING THE FLOW OF OXYGEN AND NUTRIENTS NEEDED TO MAINTAIN THE FUNCTIONING OF BRAIN CELLS . WHEN A BLOOD CLOT HAS OCCLUDED VESSEL AND REDUCES CEREBRAL BLOOD FLOW TO A SPECIFIC REGION OF THE BRAIN.

The progesteron

e has the function of inhibiting de AQP-4 proteins

neuroprotector

AQP-4 is the responsable of

edema cerebral. That increase in a

isquemia situation.

OBJETIVE

• THIS EXPERIMENT AIMED TO FURTHEREXPLORE THE MECHANISM OF PROG IN THETREATMENT OF CEREBRAL ISCHAEMIC INJURY FROM AQP-4 EXPRESSION ENCEPHALOEDEMA AND BBB PERMEABILITY

MATERIALES Y METODOS •ANIMALES : SE USARON 72 RATAS

WISTAR DE 7 DÍAS DE EDAD, ESTAS SE DIVIDIERON EN 3 GRUPOS (GRUPO CONTROL, GRUPO CON HIPOXIA-ISQUEMIA, GRUPO TRATADO CON FÁRMACOS)

LAS RATAS QUE FUERON TRATADAS CON FÁRMACOS SE LES INYECTO PROGESTERONA DE MANERA INTRAPERITONEAL A 0.5 G/L A DOSIS DE 8 MG/ KG. DESPUÉS DE LA HIPOXIA, LAS RATAS EN EL GRUPO CONTROL Y HI FUERON INYECTADOS CON SOLUCIÓN SALINA.

PERMEABILIDAD •SE DETECTO PERMEABILIDAD DE BBB ( BLOOD-BRAIN BARRIER) GRACIAS AL AZUL DE EVANS(EB) ,EL CORAZÓN FUE INYECTADO CON SOLUCIÓN SALINA 20 G/L EB ( 20 MG / KG). EL TEJIDO CEREBRAL IZQUIERDO SE PESÓ . SE DETECTÓ LA ABSORBANCIA POR ESPECTROFOTOMETRÍA. EL CONTENIDO DE EB SE CALCULÓ EN LA MUESTRA DE MEDICIÓN .

• ¿PARA QUÈ?• SE UTILIZO PARA LOGRAR

SOMETER LAS RATAS A LA HIPOXIA Y POSTERIORMENTE A

UNA ISQUEMIA.

Posición supino +

inhalación de anestesia

Piel del cuello

cortada.

Separa carótida de

el ECM

37ºc + 2-3 hrs+ 8% O2 + 92% N2

Modelos con hipoxia

ESTABLECIMIENTO DEL MODELO

MATERIALES Y METODOS •INMUNOHISTOQUIMICA • LA INMUNOHISTOQUIMICA ES LA IDENTIFICACIÓN DE UN TEJIDO

POR MEDIO DE UNA INTERACCIÓN ENTRE UN ANTÍGENO Y UN ANTICUERPO, DONDE EL ANTICUERPO HA SIDO MARCADO, LA CÉLULA SE COLOREA PARA DEMOSTRAR LA PRESENCIA Y LOCALIZACIÓN CELULAR DE UNA MOLÉCULA DE INTERÉS

EN LUGAR DE UN ANTICUERPO PRIMARIO ,SE AÑADIÓ TAMPÓN DE FOSFATO AL GRUPO DE CONTROL NEGATIVO, LAS CÉLULAS POSITIVAS SE EXPRESARON EN LA PROTEÍNA AQP.

El cerebro fue segmentado Se fijo durante la noche en 10 % de formalina , fue deshidratado, corteado , desparafinado , y conservados en un refrigerador a 4° C.

DETERMINACIÓN DEL CONTENIDO DE AGUA

Muerte de la rataIncubación del

cerebro 80% del tejido cortical en

vidrio.

Se pone electrotérmico 110

° C caja seca a temperatura constante a

hornear durante 48 h.

Peso seco

Contenido de agua del cerebro =

[(peso húmedo peso seco) / peso húmedo] x 100%.

WESTERN BLOT • FUNDAMENTO:

• PRUEBA UTILIZA PARA DETECTAR LA RELACIÓN ANTÍGENO-ANTICUERPO;

NON USA SONDAS.• USA LA TÈNICA DE SDS-PAGE: SDS -

FILTRO INCUBACIÓN CON ACRESULTADO.

Corteza Lisado de proteínas

centrifugación

Sobrenadante ácido

bicinconınico

Electroforesis en gel de

poliacrilamida

Anti-rata AQP-4 y

anticuerpo primario β-

actina

Prueba t.

Análisis de correlación=

P <0,05

RESULTADOS Los resultados inmunohistoquímicos demostraron que AQP- 4 células positivas eran redondos u ovalados . La expresión AQP- 4 en la corteza cerebral de la HI grupo fue significativamente superior a la de el grupo de control después de la hipoxia de 6 , 24 y 72h . expresión de AQP - 4 en la corteza del grupo de drogas fue significativamente menor que la del grupo de HI después de la hipoxia de 6 , 24y 72 h

ACTOR Wath say yes or no Zhou C, Tu J, Zhang Q, Lu D, Zhu Y, Zhang W,Yang F, Brann DW, Wang R

The change in microenvironment results in the up regulation of AQP-4 expression after brain injury, changes the structure of the cell membrane, increases the permeability of BBB and enhances the water permeability

yes

Sun B, Chen L, Wei X, Xiang Y, Liu X, Zhang X.

proved that AQP-4 gene knockout can significantly alleviate cerebral edema caused by acute water intoxicationor ischaemic stroke, suggesting that AQP-4performs a key function in brain tissue edema, restrains AQP-4 expression, and inhibits or alleviates cerebral edema induced by AQP-4

yes

Author What they said? Yes or not ? - Northington FJ, Chavez-

Valdez R, Martin LJ.- Doi K, Sameshima H,

Kodama Y, Furukawa S, Kaneko M, Ikenoue TK.

BBB damage is one of the most important pathophysiological mechanisms of ischaemic brain damage. Traumatic cerebral oedema is a type of mixed oedema that is closely related to the increase in BBB permeability.

Yes.

- Zhu YM, Wang CC, Chen L, Qian LB, Ma LL, Yu J, Zhu MH, Wen CY, Yu LN, Yan M.

- Zhang X, Zhang X, Wang C, Li Y, Dong L, Cui L, Wang L, Liu Z, Qiao H, Zhu C, Xing Y, Cao X, Ji Y, Zhao K.

Studies have indicated increased AQP-4 expression in cerebral oedema caused by cerebral infarction, cerebral haemorrhage, brain tumour, inflammation, and brain injury in others’ reports.

Yes.

CONCLUSION • PROGESTERONE DECREASES THE LIKELIHOOD OF CEREBRAL EDEMA DUE TO

ISCHEMIA OCCURRED . IT CAN BE INFERRED THAT BOTH NEONATAL RATS AND THEIR MOTHERS SHARE HIGH LEVELS OF THIS HORMONE DURING PREGNANCY AND SUBSEQUENT BIRTH. A CONCLUSION MAY RESULT IN THE EVOLUTIONARY DEVELOPMENT OF NEONATAL RATS (AND OTHER ORGANISMS SHARING THE PRODUCTION OF PROGESTERONE ) REDUCE THE RISK OF DEATH IN EARLY LIFE STAGES INCLUDING DURING PREGNANCY AND IN THE FIRST WEEKS AFTER BIRTH THE SAME WAY IT CAN BE ASSUMED THAT MOTHERS MAY HAVE A SIMILAR DECREASE IN THE RISK OF DEATH FROM DAMAGE TO THE IRRIGATION SYSTEM BRAIN DURING PREGNANCY AND AFTER DELIVERY.

CONCLUSION• ALTHOUGH AQP - 4 EXPRESSION IS NECESSARY IN BRAIN TISSUE BUT IT CAN ALSO BE HARMFUL

IN SOME CASES , SINCE THIS ( AQP -4 ) PRODUCES AN INCREASE IN CEREBRAL EDEMA AS IT INCREASES THUS CAN BE TARGETED INVESTIGATIONS TREATMENT IN DIFFERENT CASES AS IS THE ISQUEMIA TO PRE AND POST-OPERATIVE TREATMENTS.

• AQP4 PARTICIPANT IS IMPORTANT CAUSE OF CEREBRAL EDEMA, ALLOWING INCREASED BLOOD BRAIN BARRIER PERMEABILITY PROMOTING THE AGU ENTER HER. • HYPOXIA IS A PROCESS THAT PROMOTES THE CEREBRAL EDEMA AS IT INCREASES THE

EXPRESSION OF AQP4 CHANNELS IN THE BB. DEPUES 6 HRS OF HYPOXIA IS THAT THIS EFFECT IS SIGNIFICANTLY INTENSIFIED.

Mapa conceptual: Verónica Peréz.

Mapa conceptual : Laura Rodríguez Duque.

GRACIAS!!