Nitric Oxide and Heart Failure

Function of Nitric Oxide in Heart Remodeling and Contractility

周洲 陆峥飞 李兴哲 肖楠 董婧 袁方

Biosynthesis of Nitric Oxide

L-arginineNOS L-citrulline

NO

NOS

NOS-I nNOS low-output

NOS-II iNOS high-output

NOS-III eNOS low-output

cNOS

Ca2+

Signal Pathway of Nitric Oxide

GTP cGMPsGC

NO cGMP-dependent protein kinases

cGMP-regulated phosphodiesterasescGMP-regulated ion channels

Pr-SH Pr-S-NONO

glutathione-dependent formaldehyde dehydrogenase

eNOS in Cardiomyocyte

caveolin-3

eNOS

sarcolemma

Ca2+

calmodulin

Akt

L-ArgNO

M-R β-R B2-R

nNOS in Cardiomyocyte

nNOS localizes to cardiac SR

nNOS coimmunoprecipitates with the RYR

nNOS influences SR Ca2+ cycling

Nitric Oxide and Cardiac Myocyte Death

Nitric oxide produced by nitroglycerin protects against cardiac myocyte necrosis promoted by vasopressin.

ARNALDO PINELLI , SILVIO TRIVULZIO, LIVIO TOMASONI, et al. CARDIAC NECROSIS MARKERS ASSOCIATED WITH LOWNITRIC OXIDE LEVELS IN THE PLASMA OF RABBITS AFTER TREATMENT WITH VASOPRESSIN: PROTECTIVE EFFECTS OF NITROGLYCERIN ADMINISTRATION. Pharmacological Research, Vol. 45, No. 6, 2002

Nitric Oxide and Cardiac Myocyte Death

L-NAME induces cardiac myocyte necrosis Pinelli, Arnaldo, et al. Low Nitric Oxide Values Associated with Low Levels of Zinc and High Levels of Cardiac Necrosis Markers Detected in the Plasma of Rabbits Treated with L-NAME. Journal of cardiovascular pharmacology, 2001

DETA/NO and SNAP induces cardiac myocyte necrosis

Takamichi Uchiyama, et al. Nitric Oxide Induces Caspase-dependent Apoptosis and Necrosis in Neonatal Rat Cardiomyocytes. J Mol Cell Cardiol 34, 1049ÿ1061 (2002)

Nitric Oxide and Cardiac Myocyte Death

Jun-ichi Oyama, Stefan Frantz, et al. Nitric Oxide, Cell Death, and Heart Failure. Heart Failure Reviews, 7, 327–334, 2002

DNA damage

Mitochondrial damage

p53 increase

MAPK activation

cGMP & ceramide

Caspase inhibition

Anti-apoptosis related gene

Scavenger of ROS

eNOS and Heart Contractility

eNOS transgenic mice with very high eNOS activity exhibit inhibitory function in cardiac myocyte contractility.

F. Brunner, P. Andrew, G. Wölkart, et al. Myocardial Contractile Function and Heart Rate in Mice With Myocyte-Specific Overexpression of Endothelial Nitric Oxide Synthase. Circulation 2001;104;3097-3102

eNOS and Heart Contractility

eNOS transgenic mice with low eNOS activity improvement exhibit stimulatory function in cardiac myocyte contractility.Jun Ren, Xiaochun Zhang, Glenda I. Scott,et al. Adenovirus Gene Transfer of Recombinant Endothelial Nitric Oxide Synthase Enhances Contractile Function in Ventricular Myocytes. J Cardiovasc Pharmacol 2004;43:171–177

eNOS and Heart Contractility

eNOS transgenic mice with very high eNOS activity exhibit inhibitory function in cardiac myocyte contractility stimulated by isoprenaline.

Paul B. Massion, Chantal Dessy, Fanny Desjardins, et al.Cardiomyocyte-Restricted Overexpression of Endothelial Nitric Oxide Synthase (NOS3) Attenuates -Adrenergic Stimulation and Reinforces Vagal Inhibition of Cardiac Contraction. Circulation 2004;110;2666-2672;

eNOS and Heart Contractility

β3R

eNOS

sGC

L-Ca

Β1/2R

cAMP

NO

cGMP

GTPPKG

cGMP-stimulated cAMP-phosphodiesterase

PDE5

NO

RYR

Ca2+

nNOS and Heart Contractility

nNOS inhibited or knocked out mice exhibit a significant enhance in cardiomyocyte contractility.

Claire E. Sears, Simon M. Bryant, Euan A. Ashley,et al. Cardiac Neuronal Nitric Oxide Synthase Isoform Regulates Myocardial Contraction and Calcium Handling. Circ. Res. 2003;92;52-59;

nNOS and Heart Contractility

nNOS inhibited or knocked out mice exhibit a significant enhance in cardiomyocyte contractility stimulated by isoproterenol.

Euan A. Ashley, Claire E. Sears, Simon M. Bryant, et al. Cardiac Nitric Oxide Synthase 1 Regulates Basal and beta-Adrenergic Contractility in Murine Ventricular Myocytes. Circulation 2002;105;3011-3016

nNOS and Heart Contractility

Claire E. Sears, Simon M. Bryant, Euan A. Ashley,et al. Cardiac Neuronal Nitric Oxide Synthase Isoform Regulates Myocardial Contraction and Calcium Handling. Circ. Res. 2003;92;52-59;

Nitric Oxide and Cardiomyocyte Lusitropy

NO sGC cGMP PKG

Myosin light chain phosphatase

myofilament Ca2+ sensitivity

troponin I phosphorelation

Cardiomyocyte lusitropy

iNOS in Heart Failure

iNOS is highly expressed in compensated or decompensated failed heart. The activity is significantly higher than normal.

iNOS in Heart Failure

Specific inhibitor of iNOS 1400W can significantly enhance the contractility of compensated or decompensated mice hearts but don’t show any difference with sham-treated mice.

iNOS in Heart Failure

A B C

D

Four indexes of cardiac myocyte function of decompensated heart failure can be improved by 1400W. A, Rate of [Ca2]i activation; B, rate of [Ca2]i relaxation; C, myocyte shortening velocity; and D, myocyte relaxation velocity.Olga Gealekman, Zaid Abassi, Irit Rubinstein, et al. Role of Myocardial Inducible Nitric Oxide Synthase in Contractile Dysfunction and –Adrenergic Hyporesponsiveness in Rats With Experimental Volume-Overload Heart Failure. Circulation 2002;105;236-243

nNOS in Heart Failure

In heart failure cardiac myocyte, nNOS is greatly upregulated and translocated to the sarcolemma, binding to cavelolin-3.

nNOS in Heart Failure

After stimulated by beta-adrenin, failed hearts exhibit significant weak systole and diastole function, which can be reversed by L-NVIO.

Jennifer K. Bendall, Thibaud Damy, Philippe Ratajczak, et al. Role of Myocardial Neuronal Nitric Oxide Synthase–Derived Nitric Oxide in -Adrenergic Hyporesponsiveness After Myocardial Infarction–Induced Heart Failure in Rat. Circulation 2004;110;2368-2375;

•Part 2

nNOS in Heart Failure

Nitric oxide and cardiac myocyte apoptosis

Stress-responsive cytokines induceStress-responsive cytokines induce cardiac myocyte apoptosis cardiac myocyte apoptosis

To raise endogenous myocyte NO levels, we exposed neonatal rat ventricular cardiac myocytes to a mixture of macrophage-derived inflammatory cytokines (IL-1β, TNF- α, IFN-γ ) for periods from 1 to 5 days, and evaluated the cultures for morphologic and biochemical evidence of apoptosis.

NO involved

Apoptosis induced by IL-1β requires NO production.

• Cardiac myocytes exposed to IL-1β underwent extensive cell death that was completely blocked by inclusion of the non-selective NOS inhibitor,

L-NMMA (solid grey line).

L-NMMA: 非选择性NO 合酶抑制剂

NO as a bivalent regulator of cardiac myocyte apoptosis

Low dose NO preconditioning will ameliorate peroxynitrite induce apoptosis.

HO-1(heme oxygenase-1) is the stress-inducible molecules and strongly induced by variety stimuli, such as heme, heavy metal, inflammatory cytokines, and NO donors in VSMC.

heme oxygenase-1

血红素加氧酶

SNP was used

Sodium nitroprusside is a donor of NO that vasodilates by formation of cyclic GMP in vascular tissue.

Rat VSMCs were treated with the NO donor SNP, and NO production was estimated by measuring the stable oxidized product nitrite.

Concentration dependent

SNP treatment increased the amount of NO in a concentration dependent manner. SNP did not affect cell viability up to 0.5mM, however, it significantly decreased cell viability from 0.7mM and thereafter in a concentration dependent manner.

Apoptosis was reversed

Preconditioning of low concentration NO protects VSMC from high concentration NO-induced cell death.

Preconditioning using low SNP-induced HO-1 expression.

OH – 1 was induced

OH-1 protects from subsequent high SNP-induced cell death.

Role of OH-1

SnPP ：HO-1 inhibitor

Effects of high SNP on the p38 MAP kinase activation in VSMCs.

Effects of HO-1 induced by low SNP on the p38 MAP kinase activation in VSMCs.

Apoptosis pathyway 1

Effect of low SNP on the regulation of ΔΨm and caspase-3 activity.

Apoptosis pathyway 2

Effect of low SNP on the regulation of Bcl-2, Bax, and Apaf-1 activity.

Apoptosis pathyway 3