吡非尼酮逆转博来霉素诱导大鼠PIF的疗效及对TGF-β1、caspase-3的影响

Effect of pirfenidone on bleomycin-induced PIF in rats and its effect on TGF-β1 and caspase-3

目的探讨吡非尼酮治疗博来霉素诱导大鼠肺纤维化(PIF)的效果及对大鼠肺组织中转化生长因子β1(TGF-β1)、凋亡相关蛋白caspase-3的影响。方法选取SPF级成年SD大鼠72只,采用随机数字表法分为空白组(等量生理盐水处理)、模型组(等量生理盐水处理)、阳性对照组(强的松0.6 mg/100 g/d)、实验组(500 mg/kg/d)各18只,连续干预4周,检测各组大鼠血清中肺纤维化相关指标,采用免疫组化染色检测并肺组织中TGF-β1蛋白、caspase-3蛋白的表达程度,采用RT-PCR方法检测TGF-β1 mRNA表达程度。结果模型组大鼠的血清HA、LN、PCⅢ、Ⅳ-C、肺泡灌洗液中IL-6、IL-8水平显著的高于空白组(P<0.05);阳性对照组和实验组的血清HA、LN、PCⅢ、Ⅳ-C、肺泡灌洗液中IL-6、IL-8水平显著的低于模型组(P<0.05);模型组大鼠的肺组织中SOD、GSH-px显著的高于空白组(P<0.05),MDA、iNOS显著的高于空白组(P<0.05);阳性对照组和实验组的肺组织中SOD、GSH-px显著的高于模型组(P<0.05),MDA、iNOS显著的低于模型组(P<0.05);模型组大鼠的TGF-β1 mRNA表达水平显著的高于空白组(P<0.05);阳性对照组和实验组的肺组织TGF-β1 mRNA表达水平低于模型组(P<0.05);模型组大鼠的TGF-β1蛋白、caspase-3蛋白表达水平显著的高于空白组(P<0.05);阳性对照组和实验组的肺组织TGF-β1蛋白、caspase-3蛋白表达水平低于模型组(P<0.05)。结论吡非尼酮治疗博来霉素诱导的大鼠PIF具有一定的效果,主要与增强抗氧化能力,降低血清中纤维化指标、炎症反应程度、TGF-β1和caspase-3表达有关。

Objective To investigate the effect of pirfenidone on bleomycin-induced pulmonary fibrosis（ PIF）in rats and the changes of TGF-β1 and caspase-3 in rats. Methods 72 SPF-grade adult SD rats were randomly divided into the control group（ saline treatment）,the model group（ saline treatment）,the positive control group（ prednisone0. 6 mg/100 g/d）,and the experimental group（ 500 mg/kg/d） with 18 mice in each group,for 4 successive weeks.The related indexes of pulmonary fibrosis,including the expression of TGF-beta 1 protein and caspase-3 protein in lung tissue,were detected by immunohistochemical staining. RT-PCR was used to detect TGF-β1 mRNA expression. Results The levels of serum HA,LN,PC Ⅲ,Ⅳ-C,IL-6 and IL-8 in bronchoalveolar lavage fluid were significantly higher in the model group than in the control group（ P〈0. 05）. The levels of serum HA,LN,PC Ⅲ,Ⅳ-C,and IL-6 and IL-8 in bronchoalveolar lavage fluid were significantly lower in the positive control group and the experiment group than in the model group（ P〈0. 05）. SOD and GSH-px were significantly higher in the model group than the control group,and they were obviously higher in the positive control group and the experiment group than in the model group（ P〈0. 05）. MDA and iNOS were significantly higher in the model than the control group,and they were significantly lower in the positive control group and the experiment group than in the model group（ P〈0. 05）. The expression of TGF-β1 mRNA was significantly higher in the model group than in the control group,and it was lower in the positive control group and the experimental group than in the model group（ P〈0. 05）. The positive expression of TGF-β1 protein and Caspas-3 was significantly higher in the model group than in the control group,and it was lower in the positive control group and the experimental group than in the model group（ P〈0. 05）. Conclusion Pirfenidone has certain effect in treatment of PIF rats induced by bleomycin,mainly through decreasing serum fibrosis and inflammation,enhancing the antioxidant level and reducing the expression of caspase-3 and TGF-β1.