清开颗粒对急性肝衰竭小鼠肝细胞凋亡和Caspase-3表达的影响

目的:研究急性肝衰竭小鼠肝细胞凋亡和Caspase-3表达的变化及清开颗粒对其影响,探讨清开颗粒防治急性肝衰竭的作用机制。方法:以Ga1N LPS染毒昆明小鼠造成急性肝衰竭动物模型。分别以大、中、小剂量清开颗粒进行干预,并以大、中、小剂量促肝细胞生长颗粒(HGF)作为阳性对照,采用TUNEL染色观察肝细胞凋亡,RT-PCR检测Caspase-3 mRNA表达,采用免疫印迹方法(Western Blot)检测Caspase3表达。结果:清开颗粒各组凋亡细胞数均较HGF各组明显减少(P<0.05);清开颗粒中、高剂量组Caspase-3 mRNA表达明显低于HGF低、中剂量组(P<0.05);清开颗粒各组活化的Caspase3表达较HGF低、中剂量组显著降低(P<0.05)。结论:清开颗粒能减少内毒素肝损伤小鼠肝细胞凋亡、下调肝细胞Caspase-3的表达,提示其抑制肝细胞凋亡可能是其防治急性肝衰竭的机制之一。

Effect of "Qingkai Granule" on Hepatocyte Apoptosis and Caspase-3 Expression in Liver Tissue of Mice with Acute Hepatic Failure

Objective:To investigate hepatic cells apoptosis and the expression of caspase-3 in rats with acute hepatic failure and the action mechanism of "Qingkai Granule" (QKG) against it. Methods:Mouse model of acute hepatic failure was established by intraperitoneal injection of Ga1N and LPS. After modeling, the mice were given high dose, middle dose and low dose of QKG, and high dose, middle dose and low dose of hepatocyte growth-promoting factors(HGF) respectively. Caspase-3 mRNA expression in liver tissue were determined by RT-PCR array. Caspase-3 protein expression in liver tissue was detected by Western Blot analysis. Hepatic cells apoptosis was detected by TUNEL and HE staining. Results:The number of apoptotic hepatic cells in mice treated with QKG was significantly lower than in mice treated with HGF, P < 0.05. Compared with those in mice treated with low and middle doses of HGF, the expressions of Caspase-3 mRNA in mice given middle and high doses of QKG were much lower, P < 0.05, and the expressions of cleaved-caspase3 protein in mice given all dose of QKG were also significantly lower, P<0.05. Conclusion:QKG may alleviate endotoxin-induced liver injury and hepatic cells apoptosis, and may delay caspase-3 mRNA expression, which may be one of the potential mechanisms of preventing and treating acute hepatic failure.