二至丸有效成分对损伤后肝细胞再生障碍大鼠肝细胞凋亡及胆红素水平的影响

Influence of effective components of Erzhi Pill on hepatocyte apoptosis and bilirubin level of rats with liver cell aregeneratory after injury

目的:探索二至丸有效成分对损伤后肝细胞再生障碍大鼠肝细胞凋亡及胆红素水平的影响。方法:将40只Wistar大鼠随机分为4组(即正常组、模型组、齐墩果酸+红景天苷组、雷帕霉素组),除正常组仅行肝部分切除术(PHx)外,其余各组建立损伤后肝细胞再生抑制复合模型(PHx+2AAF),给药组分别予齐墩果酸+红景天苷和雷帕霉素治疗性给药7d后处死。采用自动生化分析仪检测血清总胆红素(TBIL)、直接胆红素(DBIL)水平,并用流式Annexin V+PI法检测肝细胞凋亡水平、Brd U流式检测肝细胞周期及Caspase-3表达。结果:与模型组比较,齐墩果酸+红景天苷组大鼠血清TBIL水平显著下降(P<0.01),而DBIL水平显著升高(P<0.05),同时肝坏死细胞水平、早期和晚期凋亡水平显著降低(P<0.01,P<0.05),肝活性细胞水平明显升高(P<0.05),肝细胞处于G0/G1期及S期细胞增殖明显升高(P<0.01,P<0.05),肝细胞Caspase-3表达水平显著降低(P<0.01)。结论:二至丸有效成分明显降低损伤后肝细胞再生障碍大鼠胆红素的分泌,抑制肝细胞凋亡和Caspase-3的活化。

Objective： To explore the influence of the effective components of Erzhi Pill on hepatocyte apoptosis and bilirubin level of rats with liver cell aregeneratory after injury. Methods： Forty Wistar rats were randomly divided into four groups（normal group, model group, oleanolic acid＋salidroside group and rapamycin group）. After the model of partial hepatectomy（PHx） were established in the normal group, the complex models of liver cells regenerative inhibition after injury（PHx＋2AAF） were established in the rest of groups, and the rats in oleanolic acid＋salidroside group were killed after therapeutic administration 7 days later. The levels of serum total bilirubin（TBIL） and direct bilirubin（DBIL） were tested by automatic biochemical analyzer, and liver cells apoptosis level was tested by the method of flow cytometry Annexin V＋PI, and liver cells cycle and caspase-3 expression were tested by Brd U flow cytometry. Results： Compared with the model group, the level of serum TBIL in the rats of oleanolic acid＋salidroside group decreased significantly（P〈0.01）, while the level of DBIL increased significantly（P〈0.05）. Meanwhile, the level of liver cells necrosis, early and late apoptotic level decreased significantly（P〈0.01, P〈0.05）, and the cell proliferation when liver cells were in G0/G1 phase and S phase increased evidently（P〈0.05, P〈0.01）; the level of Caspase-3 decreased significantly（P〈0.01）. Conclusion： The effective components of Erzhi Pill could decrease the secretion of bilirubin in the rats with liver cell aregeneratory after injury, and inhibit apoptosis of liver cell and the activation of Caspase-3.