盐酸羟考酮通过JNK/p38MAPK信号通路调控异丙肾上腺素诱导的心肌细胞凋亡的机制

目的探究盐酸羟考酮对异丙肾上腺素(ISO)诱导的心肌细胞凋亡的影响及其作用机制。方法体外培养心肌细胞H9c2,设置对照组、ISO组、盐酸羟考酮1μmol/L组、盐酸羟考酮5μmol/L组、盐酸羟考酮10μmol/L组、ISO+SB203580〔c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶(JNK/p38MAPK)信号通路阻断剂〕组。噻唑蓝(MTT)法检测心肌细胞存活率;流式细胞学法检测各组心肌细胞的凋亡率;Western印迹检测心肌细胞的B淋巴细胞瘤(Bcl)-2、B淋巴细胞瘤-2相关蛋白(Bax)及JNK/p38MAPK信号通路相关蛋白的表达;测定培养细胞上清液乳脱氢酶(LDH)、肌酸激酶(CK)含量。结果与对照组比较,ISO组心肌细胞存活率降低,各给药组心肌细胞存活率明显升高(P<0.05);与对照组比较,ISO组心肌细胞的凋亡率明显升高(P<0.05),而盐酸羟考酮可显著降低细胞凋亡率(P<0.05);与ISO组比较,盐酸羟考酮10μmol/L组LDH、CK水平及Bax、磷酸化(p)-JNK、p-p38MAPK表达水平显著降低(P<0.05),Bcl-2表达水平显著升高(P<0.05);阻断JNK/p38MAPK信号通路可显著增加细胞存活率(P<0.05),降低细胞凋亡率(P<0.05),促进Bcl-2表达(P<0.05),而抑制Bax表达(P<0.05)。结论盐酸羟考酮对ISO诱导的心肌细胞损伤具有抗凋亡作用,其作用机制可能与抑制JNK/p38MAPK信号通路的活化有关。

Effect of oxycodone hydrochloride on myocardial apoptosis induced by isoproterenol via JNK/p38MAPK signaling pathway

Objective To investigate the effect of oxycodone hydrochloride on isoproterenol(ISO)-induced cardiomyocyte apoptosis and its mechanism.Methods Cultured cardiomyocytes H9c2 in vitro,control group,ISO group,oxycodone hydrochloride 1μmol/L group,oxycodone hydrochloride 5μmol/L group,oxycodone hydrochloride 10μmol/L group,and ISO+SB203580(JNK/p38MAPK signaling pathway blocker)group were set.Myocardial cell survival rate was measured by MTT assay.The apoptosis rate of cardiomyocytes in each group was detected by flow cytometry.Western blot was used to detect the expression of Bcl-2,Bax and JNK/p38MAPK signaling pathway-related proteins in cardiomyocytes.The LDH and CK contents of the cultured cell supernatant were measured.Results Compared with the control group,the survival rate of myocardial cells in the ISO group was decreased,and the survival rate of cardiomyocytes in each drug-administered group was significantly increased(P<0.05).Compared with the control group,the apoptotic rate of myocardial cells in the ISO group was significantly increased(P<0.05),while oxycodone hydrochloride significantly decreased the apoptosis rate(P<0.05).Compared with the ISO group,the levels of LDH and CK and the expression levels of Bax,p-JNK and p-p38MAPK in oxycodone hydrochloride 10μmol/L group were significantly decreased(P<0.05),and the expression level of Bcl-2 was significantly increased(P<0.05).Blocking JNK/p38MAPK signaling pathway significantly increased cell viability(P<0.05),decreased apoptosis rate(P<0.05),promoted Bcl-2 expression(P<0.05),and inhibited Bax expression(P<0.05).Conclusions Oxycodone hydrochloride has anti-apoptotic effect on ISO-induced cardiomyocyte injury,and its mechanism may be related to inhibition of JNK/p38MAPK signaling pathway activation.