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目的观察中药二草清肝汤对大鼠免疫性肝损伤的保护作用及其机制的初步探讨。方法采用卡介苗(BCG)+脂多糖(LPS)注射制备大鼠免疫性肝损伤模型,共60只大鼠,设正常组、模型组、环磷酰胺(CTX)组、二草清肝汤小剂量组、二草清肝汤大剂量组,检测不同组血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转酶(AST)和肿瘤坏死因子-α(TNF-α)、白细胞介素-12(IL-12)的变化,流式细胞仪检测肝细胞凋亡,免疫组织化学方法检测肝细胞Bax、Bcl-2蛋白的表达。结果二草清肝汤大、小剂量组和CTX组与模型组比较,ALT及AST降低明显,ALT分别为(547±76)、(576±69)、(538±72)、(642±94)U/L,AST分别为(536±72)、(585±74)、(526±71)、(627±92)U/L,差异有统计学意义(P<0.01或P<0.05)。二草清肝汤治疗组和CTX组TNF-α、IL-12与模型组比较下降显著,差异有统计学意义(P<0.05)。二草清肝汤治疗组、CTX组与模型组比较,肝细胞凋亡率下降,差异有统计学意义(P<0.01或P<0.05);二草清肝汤治疗组、CTX组Bcl-2和Bax阳性表达及Bax/Bcl-2相对比率明显低于模型组,差异有统计学意义(P<0.01或P<0.05)。结论二草清肝汤对大鼠免疫性肝损伤具有保护作用,降低血清TNF-α、IL-12水平和抑制肝细胞凋亡可能是其作用机制之一。
Objective To observe the protective effect of Ercao Qinggan Decoction on immune hepatic injury in rats and preliminary study on its mechanism. Methods The rat model of immune hepatic injury was prepared by injection of BCG + LPS. A total of 60 rats were used. The normal group, model group, cyclophosphamide (CTX) group and Ercao Qinggan decoction were given in small doses. The serum concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin in different groups were detected in large-dose group and Ercao Qinggan decoction group. The change of IL-12 was detected by flow cytometry, and the expression of Bax and Bcl-2 protein in hepatocytes was detected by immunohistochemistry. Results Compared with the model group, the levels of ALT and AST decreased significantly in Ercao Qinggan Decoction group, CTX group, and ALT group (547±76), (576±69), (538±72), (642±94). U/L and AST were (536±72), (585±74), (526±71), (627±92) U/L, respectively, with statistical significance (P<0.01 or P<0.05). The TNF-α and IL-12 levels in Ercao Qinggan decoction group and CTX group were significantly lower than those in the model group, and the difference was statistically significant (P<0.05). Compared with the model group, Ercao Qinggan decoction treatment group, CTX group and the model group, the apoptosis rate of hepatocytes decreased, the difference was statistically significant (P<0.01 or P<0.05); Ercao Qinggan Decoction treatment group, CTX group Bcl-2 The relative expression of Bax and Bax/Bcl-2 was significantly lower than that of the model group, and the difference was statistically significant (P<0.01 or P<0.05). Conclusion Ercao Qinggan decoction has protective effect on immune hepatic injury in rats. Decreasing serum TNF-α and IL-12 levels and inhibiting hepatocyte apoptosis may be one of the mechanisms.