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目的探讨复方抗纤二号抗肝纤维化的治疗机制。方法雄性Wistar大鼠分成5组,除正常对照外,余4组均腹腔注射猪血清(0.5 ml/次,2次/周,共12周)用作肝纤维化造模。抗纤二号早期治疗组B在第3周给予中药灌胃,1 ml/100 g体重,每天一次。抗纤二号晚期治疗组C在第9周给予中药灌胃,1 ml/100 g体重,每天一次。γ-干扰素治疗组D在第9周每天皮下注射10万单位的干扰素。模型组A和正常对照组N给等量的生理盐水灌胃。12周末杀大鼠,苏木精-伊红染色和Masson染色观察肝纤维化的形成,免疫组织化学观察平滑肌肌动蛋白(SMA)的表达。同时逆转录聚合酶链反应(RT-PCR)检测肝组织中SMA、Ⅰ、Ⅲ胶原和转化生长因子β1(TGF-β1)下游信号Smad3 mRNA的表达。结果抗纤二号治疗组B和C与模型组比较,体重较重,肝脏、脾脏变小,肝重/体重和脾重/体重降低(P<0.05)。病理学观察,抗纤二号治疗组B显著逆转了免疫性大鼠的肝纤维化。苏木精-伊红染色和Masson染色显示抗纤二号治疗组B胶原明显减少(P<0.05)。逆转录聚合酶链反应分析SMA、Ⅰ、Ⅲ型胶原和Smad3 mRNA的表达在抗纤二号治疗组均明显减少(P<0.05)。免疫组织化学观察SMA的表达在抗纤二号治疗组均明显降低,同时分析表明Smad3 mRNA与Ⅰ、Ⅲ胶原mRNA存在正相关(r=0.890)。结论抗纤二号能逆转免疫性肝纤维化,这是由于能部分抑制肝星状细胞的增殖和抑制肝纤维化有关的细胞因子TGF-β1下游信号Smad3的表达。
Objective To explore the therapeutic mechanism of compound anti-fibrosis II anti-fibrosis. Methods Male Wistar rats were divided into 5 groups. In addition to the normal controls, the remaining 4 groups were injected intraperitoneally with pig serum (0.5 ml/time, twice/week for 12 weeks) as liver fibrosis model. Anti-fibral early-treatment group B was given intragastric administration of gavage at the third week, 1 ml/100 g body weight once a day. The anti-fibrosis 2 advanced treatment group C was given intragastric administration at the 9th week, 1 ml/100 g body weight once a day. In the IFN-gamma treatment group D, 100,000 units of interferon were injected subcutaneously every day during the 9th week. Model group A and normal control group N were given intragastric administration of equal volume of normal saline. At the end of the 12th week, rats were killed, hematoxylin-eosin staining and Masson staining were used to observe the formation of hepatic fibrosis. The expression of smooth muscle actin (SMA) was observed by immunohistochemistry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of SMA, I, III collagen and transforming growth factor β1 (TGF-β1) downstream signal Smad3 mRNA in liver tissue. Results Compared with the model group, the anti-fibrosis treatment group B and C had heavier body weight, smaller liver and spleen, lower liver weight/weight and spleen weight/weight (P<0.05). Pathological observations that anti-fibrosis treatment group B significantly reversed hepatic fibrosis in immunized rats. Hematoxylin-eosin staining and Masson staining showed that collagen B in the anti-fibrosis treatment group was significantly reduced (P<0.05). Reverse transcriptase polymerase chain reaction analysis of the expression of SMA, type I, type III collagen and Smad3 mRNA was significantly reduced in the anti-fibrosis treatment group (P<0.05). Immunohistochemical observation of SMA expression in the anti-fibrosis treatment group were significantly reduced, while analysis showed that Smad3 mRNA and collagen I, III there is a positive correlation (r=0.890). Conclusion Anti-fibrosis can reverse the immune fibrosis, which is due to the partial inhibition of the proliferation of hepatic stellate cells and the suppression of hepatic fibrosis related cytokine STGF3 downstream signaling TGF-β1 expression.