山奈酚对大鼠心肌细胞H9c2凋亡的影响
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摘要
目的:探讨山奈酚对大鼠心肌细胞凋亡的影响及其机制。方法:通过LPS体外诱导H9c2细胞,完成心肌细胞凋亡模型的建立,以0、5、25、50μmol/L山奈酚和(或) LPS对心肌细胞进行干预,并采用CCK-8法检测其对细胞的毒性,细胞凋亡率采用TUNEL染色进行检测,最后通过Western blot法检测各组细胞TLR4/NF-κB信号通路中P65、p-P65、p-Iκbα、Iκbα、TLR4蛋白的表达情况以及凋亡相关蛋白cleaved Caspase(c-Caspase)-8、c-Caspase-9、cCaspase-3的表达情况。结果:LPS组细胞凋亡率较对照组升高,LPS+山奈酚组细胞凋亡率较LPS组下降(P<0.05)。LPS可使细胞活性降低,而山奈酚可提高细胞活性。LPS干预后,凋亡相关蛋白表达上调,凋亡相关信号通路TLR4/NF-κB蛋白表达增加(P<0.05);山奈酚可抑制凋亡相关蛋白表达和TLR4/NF-κB信号通路蛋白的表达(P<0.05)。结论:LPS诱导的H9c2细胞凋亡可被山奈酚所抑制,其机制与抑制TLR4/NF-κB信号通路的活化有关。
Aim:To investigate the effect of kaempferol on rat cardiomyocyte apoptosis and its mechanism.Methods:H9c2 cells were induced by LPS to establish cardiomyocyte apoptosis model in vitro.Toxicity of 0,5,25,50 μmol/L kaempferol and/or LPS to cells was detected by CCK-8 and the cell apoptotic rate was observed by TUNEL staining.The expressions of TLR4,p-Iκbα,p-P65,Iκbα and P65 in TLR4/NF-κB signaling pathway and the expressions of apoptosis-related proteins cleaved Caspase(c-Caspase)-8,c-Caspase-9 and c-Caspase-3 were detected by Western blot.Results:The cell apoptosis rate in LPS group was higher than those in normal control group and LPS + kaempferol group(P<0.05).Compared with normal control group,LPS could significantly reduce cell viability,while kaempferol could increase cell viability.After LPS intervention,the expressions of the apoptosis-related protein were up-regulated(P<0.05).Besides,the apoptosis-related signaling pathway TLR4/NF-κB was also activated(P<0.05).Kaempferol could significantly reduce apoptosis rate and it could also inhibit the activation of apoptosis protein and TLR4/NF-κB signaling pathway(P<0.05).Conclusion:Kaempferol could inhibit the apoptosis of H9c2 cells induced by LPS,and the mechanism is related to the inhibition of activation of TLR4/NF-κB signaling pathway.
Aim:To investigate the effect of kaempferol on rat cardiomyocyte apoptosis and its mechanism.Methods:H9c2 cells were induced by LPS to establish cardiomyocyte apoptosis model in vitro.Toxicity of 0,5,25,50 μmol/L kaempferol and/or LPS to cells was detected by CCK-8 and the cell apoptotic rate was observed by TUNEL staining.The expressions of TLR4,p-Iκbα,p-P65,Iκbα and P65 in TLR4/NF-κB signaling pathway and the expressions of apoptosis-related proteins cleaved Caspase(c-Caspase)-8,c-Caspase-9 and c-Caspase-3 were detected by Western blot.Results:The cell apoptosis rate in LPS group was higher than those in normal control group and LPS + kaempferol group(P<0.05).Compared with normal control group,LPS could significantly reduce cell viability,while kaempferol could increase cell viability.After LPS intervention,the expressions of the apoptosis-related protein were up-regulated(P<0.05).Besides,the apoptosis-related signaling pathway TLR4/NF-κB was also activated(P<0.05).Kaempferol could significantly reduce apoptosis rate and it could also inhibit the activation of apoptosis protein and TLR4/NF-κB signaling pathway(P<0.05).Conclusion:Kaempferol could inhibit the apoptosis of H9c2 cells induced by LPS,and the mechanism is related to the inhibition of activation of TLR4/NF-κB signaling pathway.
引文
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[14]ZHAO JJ,HU YW,HUANG C,et al.Dihydrocapsaicin suppresses proinflammatory cytokines expression by enhancing nuclear factor IA in a NF-κB-dependent manner[J].Arch Biochem Biophys,2016,604:27
[15]YU Y,LI S,LI Y,et al.Fibroblast growth factor 21(FGF21)ameliorates collagen-induced arthritis through modulating oxidative stress and suppressing nuclear factorkappa B pathway[J].Int Immunopharmacol,2015,25(1):74
[2]陈育华,周克元,袁汉尧.山奈酚药效的研究进展[J].广东医学,2010,31(8):1064
[3]于雪,付帮泽,郭淑贞,等.山奈酚和芒柄花黄素对缺氧/复氧条件下H9C2心肌细胞活性氧水平的影响[J].辽宁中医杂志,2017,44(1):111
[4]LIU Y,GAO L,GUO S,et al.Kaempferol alleviates angiotensinⅡ-induced cardiac dysfunction and interstitial fibrosis in mice[J].Cell Physiol Biochem,2017,43(6):2253
[5]李异,王慷慨,蒋永芳,等.心型脂肪酸结合蛋白对脂多糖所致心肌细胞损伤的保护作用[J].中南大学学报(医学版),2015,40(5):457
[6]黄锦达,刘翠,郑贵浪,等.胰岛素对脂多糖诱导的H9c2心肌细胞损伤的保护作用及其机制[J].中华实用儿科临床杂志,2016,31(6):429
[7]孔令希.山奈酚抗动脉粥样硬化作用机理探讨及其药代动力学研究[D].重庆:重庆医科大学,2014.
[8]周明杰,刘立群.山奈酚预处理对大鼠心脏缺血再灌注损伤的影响[J].潍坊医学院学报,2015,37(3):168
[9]XIAO J,SUN GB,SUN B,et al.Kaempferol protects against doxorubicin-induced cardiotoxicity in vivo and in vitro[J].Toxicology,2012,292(1):53
[10]倪青.糖尿病心肌病心室重塑与细胞凋亡的关系及中医药干预前景[J].医学研究杂志,2016,45(11):1
[11]ZHUANG YT,XU DY,WANG GY,et al.IL-6 induced lncRNA MALAT1 enhances TNF-αexpression in LPS-induced septic cardiomyocytes via activation of SAA3[J].Eur Rev Med Pharmacol Sci,2017,21(2):302
[12]ZHANG Z,LI YM,SHENG CQ,et al.Tanshinone IIA inhibits apoptosis in the myocardium by inducing microRNA-152-3p expression and thereby downregulating PTEN[J].Am J Transl Res,2016,8(7):3124
[13]LI B,CHI RF,QIN FZ,et al.Distinct changes of myocyte autophagy during myocardial hypertrophy and heart failure:association with oxidative stress[J].Exp Physiol,2016,101(8):1050
[14]ZHAO JJ,HU YW,HUANG C,et al.Dihydrocapsaicin suppresses proinflammatory cytokines expression by enhancing nuclear factor IA in a NF-κB-dependent manner[J].Arch Biochem Biophys,2016,604:27
[15]YU Y,LI S,LI Y,et al.Fibroblast growth factor 21(FGF21)ameliorates collagen-induced arthritis through modulating oxidative stress and suppressing nuclear factorkappa B pathway[J].Int Immunopharmacol,2015,25(1):74