姜黄素联合黄芩苷对乙醇诱导大鼠肝细胞凋亡的干预作用研究

Interventional effect of curcumin combined with baicalin on ethanol-induced hepatocyte apoptosis in rats

目的探讨姜黄素和黄芩苷对乙醇诱导大鼠肝细胞凋亡的保护作用及其潜在机制。方法 50只健康雄性Wistar大鼠随机分为正常对照组、乙醇模型组(50%v/v)、姜黄素干预组(50 mg/kg·bw)、黄芩苷干预组(50 mg/kg·bw)及姜黄素和黄芩苷联合干预组。TUNEL染色观察大鼠肝细胞凋亡情况,RT-qPCR检测大鼠肝CHOP、GRP78、ASK1、MKK3、Caspase-3和Caspase-12的mRNA表达水平,Western blot检测大鼠肝CHOP、GRP78、ASK1、p-ASK1、MKK3、p-MKK3、Caspase-3和Caspase-12的蛋白表达水平。结果与对照组相比,乙醇模型组肝组织TUNEL阳性细胞数、CHOP、GRP78、Caspase-3和Caspase-12的mRNA表达水平及CHOP、GRP78、p-ASK1、p-MKK3、Caspase-3和Caspase-12的蛋白表达水平均上调(P<0.05)。联合干预组TUNEL阳性细胞数、CHOP、GRP78、p-MKK3、Caspase-3和Caspase-12的蛋白表达水平均低于乙醇模型组及姜黄素和黄芩苷单独干预组(P<0.05),CHOP和GRP78的mRNA表达水平低于乙醇模型组(P<0.05),p-ASK1水平和Caspase-3 mRNA表达水平低于乙醇模型组和姜黄素干预组(P<0.05),Caspase-12 mRNA表达水平低于乙醇模型组和黄芩苷干预组(P<0.05)。此外,姜黄素和黄芩苷联用对GRP78和Caspase-3 mRNA表达及p-ASK1/ASK1和p-MKK3/MKK3的比值存在协同交互作用,差异均有统计学意义(P<0.05)。结论姜黄素和黄芩苷可能通过抑制内质网应激凋亡通路而减轻乙醇诱导的大鼠肝细胞过度凋亡。

Objective To investigate the protective effect of curcumin and baicalin on ethanol-induced hepatocyte apoptosis and its potential mechanism. Methods Fifty wistar rats were randomly divided into control group, ethanol group(50% v/v), curcumin group(50 mg/kg·bw), baicalin group(50 mg/kg·bw), and curcumin+baicalin group. TUNEL staining was used to observe the apoptosis of rat hepatocytes. RT-qPCR was used to detect the mRNA levels of CHOP, GRP78, ASK1, MKK3, Caspase-3, and Caspase-12 in rat liver. Western blot was used to detect the protein levels of CHOP, GRP78, ASK1 p-ASK1, MKK3, p-MKK3, Caspase-3 and Caspase-12 in rat liver. Results Compared with the control group, the number of tunel positive cells, the mRNA levels of CHOP, GRP78, Caspase-3 and Caspase-12 and the protein levels of CHOP, GRP78, p-ASK1, p-MKK3, Caspase-3 and Caspase-12 in the ethanol group were up-regulated(P<0.05). In the combination group, the number of tunel positive cells, the protein contents of CHOP, GRP78, p-MKK3, Caspase-3 and Caspase-12 were lower than that in the ethanol group and each drug alone intervention group(P<0.05);CHOP and GRP78 mRNA levels were lower than that in the ethanol group(P<0.05);p-ASK1 level and Caspase-3 mRNA expression were lower than that in the ethanol group and curcumin group(P<0.05);Caspase-12 mRNA expression was lower than that in the ethanol group and Baicalin group(P<0.05). In addition, curcumin combined with Baicalin had a synergistic interaction on the expression of GRP78 and Caspase-3 mRNA and the ratios of p-ASK1/ASK1 and p-MKK3/MKK3(P<0.05). Conclusion Curcumin and baicalin may reduce ethanol-induced excessive apoptosis of rat hepatocytes by inhibiting the endoplasmic reticulum stress apoptosis pathway.