The Expriment Study On The Mechanisms Of Adjusting Cytokines And Improving Heart Function

Objective: To find out the relationship among rennin-angiotensin system,oxidative stress and cytokines,and try to clarificate cytokines mechanism of valsartan and carvedilol in treating CHF by studying the role of angiotensinⅡ and H2O2 in synthesizing TNF-α,NO,MDA in culture human PBMC and neonatal rat cardiac myocyte treated with valsartan and carvedilol.Meathod: The experiment adopted density gradient centrifuging to separate PBMC of 44 patients with CHF and 20 normal person and enzyme digestion to separate neonatal rat cardiac myocyte. PBMC and cardiac myocyte were cultured for 24 hours with AngⅡand H2O2 after being pretreated with valsartan and carvedilol,then detecting TNF-αconcentration in culture supernatants by ELISA and TNF-αmRNA expression in cardiac myocyte by RT-PCR,adopting nitrate reductase to detect NO concentration and chromatometry to detect MDA concentration in culture supernatants.Result:(1) The different concentration of valsartan inhibited TNF-α excretion from PBMC induced by LPS in concentration-dependent manner. Treatment PBMC induced by LPS of patients with CHF and normal person(n=10)with valsartan (0.1μmol/L) inhibited TNF-α(324.30±80.23 pg/ml,661.77±135.96 pg/ml)excretion ；Treatment PBMC induced by LPS patients with CHF and normal person (n=10)with carvedilol (0.1μmol/L)inhibited TNF-α(266.42±96.94pg/ml,651.65±125.64pg/ml) excretion, and there were significant differences of all compared with control group(P﹤0.05). (2) The different concentration of AngⅡ,H2O2 stimulated TNF-αfrom PBMC in concentration-dependent manner. AngⅡ(0.1μmol/L) stimulated TNF-α(384.17±99.04pg/ml,256.49±52.47pg/ml) excretion from PBMC of patients with CHF and normal person (n=10); H2O2(0.1mmol/L)induced TNF-α(445.23±76.43pg/ml,341.67±66.38pg/ml) excretion from PBMC of patients with CHF and normal person (n=10),and there were significant differences of all compared with control group(P﹤0.01). Valsartan and carvedilol inhibited the role of AngⅡ and H2O2,respectively(P﹤0.05)(3)There no TNF-αmRNA expression in control group,the levels of TNF-αmRNA expression were 0.41±0.09,0.53±0.15, 0.78±0.21 in AngⅡ(0.01 μmol/L,0.1 μmol/L,1 μmol/L) groups(n=4); The levels of TNF-αmRNA expression were 0.34±0.14,0.72±0.27,0.77±0.31 in H2O2 (0.01 mmol/L,0.1 mmol/L,0.5mmol/L) groups(n=4) ,respectively, there were significant differences of all compared with control group(P﹤0.01). Valsartan and carvedilol inhibited the above role of AngⅡ and H2O2,respectively(P﹤0.05)Conclusion:(1)In a specific concentration range,valsartan and carvedilol inhibited TNF-αsynthesis from PBMC induced by LPS in concentration- dependent manner.(2)In a specific concentration range, AngⅡ,H2O2 stimulated TNF-αsynthesis from PBMC in concentration-dependent manner,which were inhibited by valsartan and carvedilol,respectively.(3)AngⅡ,H2O2 stimulated TNF-α mRNA expression,which were inhibited by valsartan and carvedilol.(4)There is internal correlation among ennin-angiotensin system,oxidative stress and cytokines.