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[目的]观察正肝方对黄曲霉毒素B1(AFB1)诱发的大鼠癌前病变肝组织细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白依赖性激酶4(CDK4)的影响。[方法]Wistar大鼠100只随机分为4组:模型、正肝方小剂量、正肝方大剂量组各30只、正常组10只。除正常组外,先后用AFB1和2-乙酰氨基芴(2-AAF)处理各组大鼠造模。正肝方大、小剂量组在造模期间,将正肝方水剂(0.6、0.3 g/ml)按10 ml/kg分别灌胃;模型组用无菌蒸馏水按10 ml/kg灌胃。8周后处死大鼠,肝组织取材。分别采用免疫组化法和RT-PCR法,检测大鼠肝组织中Cyclin D1、CDK4蛋白和mRNA表达。[结果]模型组Cyclin D1、CDK4染色阳性细胞百分率(LI)和表达强度(PU)最高,Cyclin D1、CDK4mRNA表达水平也最高(P<0.01)。正肝方能降低大鼠癌前病变肝组织Cyclin D1、CDK4蛋白和mRNA的异常高表达(P<0.01或<0.05),且呈现出一定的量效关系。[结论]正肝方能通过下调Cyclin D1、CDK4异常表达,改善细胞周期G1~S期调控点失控,从而减缓肝细胞异常增生。
[Objective] To observe the effect of Zhengganfang on the expression of Cyclin D1 and Cyclin-Dependent Kinase 4 (CDK4) in the liver of precancerous lesion induced by aflatoxin B1 (AFB1) in rats. [Methods] One hundred Wistar rats were randomly divided into 4 groups: model, Zhengganfang small dose, Zhengganfang large dose group, 30 rats and normal group, 10 rats. In addition to the normal group, rats in each group were treated with AFB1 and 2-acetylaminofluorene (2-AAF) successively. Zhenggan liver Fangda, low-dose group in the modeling period, the Zhengganfang agent (0.6,0.3 g / ml) by 10 ml / kg respectively; model group with sterile distilled water by gavage at 10 ml / kg. After 8 weeks, the rats were sacrificed and the liver tissues were drawn. Immunohistochemistry and RT-PCR were used to detect the expression of Cyclin D1, CDK4 protein and mRNA in rat liver tissue. [Results] The positive rate of Cyclin D1 and CDK4 positive cells in model group was the highest, while the expression of Cyclin D1 and CDK4 mRNA was the highest (P <0.01). Zhenggan Decoction can reduce the abnormally high expression of Cyclin D1, CDK4 protein and mRNA (P <0.01 or <0.05) in the liver tissue of precancerous lesions in rats, and shows a certain dose-effect relationship. [Conclusion] Zhenggan decoction can reduce the abnormal expression of Cyclin D1 and CDK4 and improve the control of G1 - S phase of cell cycle, which can reduce the abnormal proliferation of hepatocytes.