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目的观察肝纤维化大鼠肝脏Bcl-2、Bax、增殖细胞核抗原(PCNA)的表达及重组转化生长因子(TGF)-β1疫苗对其表达的影响,探讨肝细胞凋亡、增殖与肝纤维化的关系及重组TGF-β1疫苗对凋亡因子及PCNA表达的影响。方法将健康雄性Sprague-Dawley系大鼠30只分为正常组、肝纤维化模型组和TGF-β1疫苗治疗组,每组各10只。肝纤维化模型组以0.5%的二甲基亚硝胺(DMN)按0.2 mL/100 g的剂量腹腔内注射,TGF-β1疫苗治疗组在注射DMN的基础上,皮下注射疫苗150μg。42 d后处死,分批从各组取血清、肝脏组织,采用S-P免疫组织化学方法检测Bcl-2、Bax和PCNA的表达,并检测血ALT、AST、Alb、HA及LN。结果Bax与大鼠肝脏病理分级呈正相关,Bcl-2、Bcl-2/Bax与大鼠肝脏病理分级呈负相关,PCNA与大鼠肝脏病理分级无明显相关性。正常组大鼠肝组织中TGF-β1、Bax表达水平较低,明显低于肝纤维化模型组(P<0.01),Bcl-2表达水平与肝纤维化模型组相当,差异无统计学意义。但TGF-β1疫苗治疗组肝组织中TGF-β1表达明显下降,Bcl-2表达水平明显升高,与肝纤维化模型组相比,差异有统计学意义(P<0.05),而Bax表达水平与肝纤维化模型组相当,差异无统计学意义。肝纤维化模型组PCNA的水平较正常组高,差异有统计学意义(P<0.01),但TGF-β1疫苗治疗组PCNA的水平较肝纤维化模型组又明显提高,差异有统计学意义(P<0.01)。结论肝细胞凋亡是引起肝纤维化的可能机制之一。TGF-β1疫苗可能通过影响肝纤维化大鼠Bcl-2、Bax、PCNA的表达而影响肝细胞的凋亡和增殖,从而改善肝脏病理分级。
Objective To observe the expression of Bcl-2, Bax, proliferating cell nuclear antigen (PCNA) and the effect of recombinant TGF-β1 vaccine on hepatic fibrosis in rats and to explore the correlation between hepatocyte apoptosis, proliferation and hepatic fibrosis And the effect of recombinant TGF-β1 vaccine on apoptosis and PCNA expression. Methods Thirty healthy male Sprague-Dawley rats were divided into normal group, liver fibrosis model group and TGF-β1 vaccine treatment group, with 10 rats in each group. The liver fibrosis model group was injected intraperitoneally with 0.5% dimethylnitrosamine (DMN) at a dose of 0.2 mL / 100 g, and the TGF-β1 vaccine group was injected subcutaneously with 150 μg of vaccine on the basis of DMN injection. After 42 days, the rats were sacrificed and the serum and liver tissues were collected in batches. The expressions of Bcl-2, Bax and PCNA were detected by S-P immunohistochemistry. ALT, AST, Alb, HA and LN were detected. Results Bax was positively correlated with the pathological grade of liver in rats. Bcl-2 and Bcl-2 / Bax were negatively correlated with the pathological grade of liver in rats. There was no significant correlation between PCNA and pathological grade of liver in rats. The expression of TGF-β1 and Bax in liver tissue of normal rats was lower than that of hepatic fibrosis rats (P <0.01), and the expression of Bcl-2 was similar to that of hepatic fibrosis rats. The difference was not statistically significant. However, the expression of TGF-β1 in liver tissue of TGF-β1 vaccine group was significantly decreased and the expression of Bcl-2 was significantly higher than that in liver fibrosis model group (P <0.05), while the expression of Bax Compared with the liver fibrosis model group, the difference was not statistically significant. The level of PCNA in liver fibrosis model group was higher than that in normal group (P <0.01), but the level of PCNA in TGF-β1 vaccine group was significantly higher than that in liver fibrosis model group (P <0.01) P <0.01). Conclusion Hepatocyte apoptosis is one of the possible mechanisms of liver fibrosis. The TGF-β1 vaccine may affect hepatic cell apoptosis and proliferation by affecting the expression of Bcl-2, Bax and PCNA in rats with hepatic fibrosis, thereby improving the pathological grade of liver.