Simvastatin inhibit the expression of tumor necrosis factor-αin rat cadiocyte and the relation with reactive oxygen species

Abstract

Abstract: AIM: To investigate the effects of simvastatin (Sim) on the expression of tumor necrosis factor- α( TNF-α) in neonatal rat cardiomyocytes stimulated by lipopolysaccharide ( LPS) and approach the underlying molecular mechanisms.METHODS: The cadiocyte, as experimental model, was isolated and cultured from neonatal Sprague-Dawley rats.The expressions of TNF-αmRNA and protein level were detected by RT-PCR and ELISA.The reactive oxygen species ( ROS) levels in cadiocyte were detected by peroxide specific probe 2 ', 7 '-dichlorofluorescin diacetate (DCF-DA).The relative activity of NADPH oxidase was measured by cytochrome C reduction assay.RESULTS: The TNF-αmRNA of cadiocyte in cardiomyocytes stimulated with LPS, the expressions of TNF-α mRNA and protein level [ ( 0.29 ±0.07), ( 83.6 ± 14.5) pg/mL] were higher than those in control group [ ( 0.07 ±0.02), ( 22.1 ±7.1) pg/mL] ( P <0.01). The levels of TNF-αmRNA and protein in 1 μmol/L Sim +LPS and 10 μmol/L Sim +LPS groups were (0.18 ±0.05), ( 0.15 ±0.02) and ( 43.1 ±8.3), ( 40.1 ±7.3) pg/mL, respectively.Compared with LPS group, which significantly were decreased( P <0.01). The TNF-αprotein contents were ( 39.1 ±10.2), ( 37.9 ±8.9) pg/mL pretreatment with antioxidants Nacetylcysteine and diphenyleneiodonium, compared with LPS group, which significantly were decreased ( P < 0.01).Compared with control group, the DCF-fluorescence intensity ( 83 ±15) and the NADPH oxidase activity( 246 ±43) % in LPS group were significantly increased ( P <0.01), which were decreased by pretreatment with Sim[ ( 41 ±8), ( 120 ±17) %, P < 0.01].Compared with control group, the TNF-αprotein content was increased( 65.0 ±17.2) pg/mL, stimulated by exogenous hydrogen peroxide.The TNF-α protein content was ( 58.8 ±15.8) pg/mL pretreatment with Sim, compared with H₂O₂ group, there was no siginificant difference.Compared with Sim group, the expression of TNF-αmRNA and protein level, ROS level, NADPH oxidase activity were increased pretreatment with mevalonate and Sim.CONCLUSION: Sim probably inhibits the LPS-induced expression of TNF-αin cultured cadiocyte via modulating the activity of NADPH oxidase and subsequent production of ROS.

Key words: statins, lipopolysaccharide, inflammatory cytokines, cadiocyte, reactive oxygen species

CLC Number:

R965.2

SHANG Fu-jun, WANG Jie-pin, ZHENG Qiang-sun, LIU Xiong-tao, XUE Yu-sheng, HE Yong, ZHANG Lu-xing, LIU Hui, WANG Bin, ZHAO Lian-you. Simvastatin inhibit the expression of tumor necrosis factor-αin rat cadiocyte and the relation with reactive oxygen species[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(12): 1379-1385.

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