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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 effect of Erzhi Pill on hepatocyte apoptosis and bilirubin level in rats with hepatocellular aplasia after injury. Methods: Forty Wistar rats were randomly divided into four groups (normal group, model group, oleanolic acid + salidroside group, rapamycin group). Only hepatectomy (PHx ), And the rest of the groups were established PHX + 2AAF compound model (PHx + 2AAF). The rats in the treatment group were treated with oleanolic acid + salidroside and rapamycin respectively for 7 days and then sacrificed. Serum total bilirubin (TBIL) and direct bilirubin (DBIL) levels were detected by automatic biochemical analyzer. The apoptosis rate of hepatocytes was detected by flow cytometry with Annexin V + PI method. The cell cycle of hepatocytes and Caspase- 3 expression. Results: Compared with the model group, the levels of serum TBIL in the oleanolic acid + salidroside group were significantly decreased (P <0.01), while the levels of DBIL were significantly increased (P <0.05), while the levels of hepatic necrosis cells, (P <0.01, P <0.05), and the level of hepatic active cells was significantly increased (P <0.05). The proliferation of hepatocytes at G0 / G1 phase and S phase was significantly increased (P <0.01, P <0.05), the expression of Caspase-3 in hepatocytes decreased significantly (P <0.01). Conclusion: Erzhi Pills can effectively reduce the bilirubin secretion and inhibit the apoptosis of hepatocytes and the activation of Caspase-3 in rats with hepatocellular aplasia after injury.