云药天龙竭分期干预对肺纤维化大鼠肺组织TGF-β_(1)表达及病理变化的影响

Tianlongjie stage intervention on TGF-β_(1) level and pathological changes in lung tissue of pulmonary fibrosis rats

目的 观察天龙竭对肺纤维化(PF)大鼠肺组织中转化生长因子β_(1)(TGF-β_(1))表达的影响及病理变化,探讨其干预PF的作用机制。方法 采用博来霉素法建立大鼠PF模型。90只大鼠随机分为9组:空白对照组,模型早期组,模型晚期组,天龙竭早期干预中剂量组,天龙竭晚期干预低、中、高剂量(1.26、2.52、5.04 g·kg^(-1)·d^(-1))组,吡非尼酮早期干预组,吡非尼酮晚期干预组。早期干预组与晚期干预组分别于造模后第7天或第14天起灌胃相应药物,连续用药,观察大鼠的生存状态,并于造模后第28天取材。以SABC和RT-PCR技术检测TGF-β_(1)在肺组织中的蛋白及mRNA表达水平,经HE染色、Masson三联染色观察肺组织病理变化。结果 TGF-β_(1)表达:与模型组比较,天龙竭早期及晚期干预均可降低PF大鼠肺组织中TGF-β_(1)的mRNA及蛋白表达水平(P <0.05,P <0.01);早期干预与晚期干预中剂量组比较,存在显著性差异(P <0.05);晚期干预高、中剂量组与低剂量组比较,存在显著性差异(P <0.05)。吡非尼酮早期及晚期干预均可降低PF大鼠肺组织中TGF-β_(1)的mRNA及蛋白表达水平(P <0.05)。天龙竭早期干预与吡非尼酮早期干预比较无显著性差异(P> 0.05),天龙竭晚期干预中剂量组与吡非尼酮晚期干预比较无显著性差异(P> 0.05)。病理变化:与模型组比较,天龙竭及吡非尼酮干预均可减轻肺泡炎、胶原沉积及间质细胞增生,早期干预作用更为明显。病理形态学定量分析显示,天龙竭及吡非尼酮早期干预评分无显著性差异(P> 0.05)。结论 天龙竭能有效减缓博来霉素诱发的肺泡炎及纤维化进程,早期干预更有意义。其机制可能与抑制PF发生发展过程中重要因子TGF-β_(1)在肺组织的表达有关。

Objective To observe the effects of Tianlongjie on the expression of TGF-β_(1) and histopathological changes in lung tissue of pulmonary fibrosis(PF) rats and to explore the possible mechanism of Tianlongjie in treating PF.Methods Rat pulmonary fibrosis models were prepared by intratracheal bleomycin infusion.Ninety male Wista rats were randomized divided into 9 groups:control group(C),model early group(ME) and model late group(ML),Tianlongjie early intervention group with middle dose(TEM),Tianlongjie late intervention group with low dose group(TLL),TIANLONGJIE late intervention group with middle dose group(TLM),Tianlongjie late intervention group with high dose group(TLH)(1.26、2.52、5.04 g·kg^(-1)·d^(-1)).Pirfenidone early intervention group(PFDE) and Pirfenidone late intervention group(PFDL).Rats of early intervention group were given drug from 7th day after PF model was established,and late intervention group was given drug from 14th day.Observe the survival state of rats and select materials on the 28th day.The protein and mRNA expression of TGF-β_(1) in lung tissue were detected by SABC and Real Time PCR.HE staining and Masson triple staining of lung tissue were used to analyze the pathological morphology.Results Expression of TGF-β_(1),Compared with the model group,early and late intervention with Tianlongjie inhibit the expression of TGF-β_(1) mRNA and protein in lung tissue of PF rats(P<0.05,P<0.01),and there was a significant difference between TEM and TLM(P<0.05).Compared with TLL,the TLM and TLH were more effective(P<0.05).Early and late intervention with Pirfenidone inhibit the expression of TGF-β_(1) mRNA and protein in lung tissue of PF rats(P<0.05).There was no statistical difference between TEM and PFDE(P>0.05),and the same was true in TLM and PFDL.Pathological morphology:The pathological results showed that intervention with Tianlongjie and Pirfenidone reduce the degree of alveolitis,the deposition of collagen and interstitial cell proliferation in lung tissue.The effect of early intervention is more obvious.Based on pathomorphology quantitative analysis,there was no statistical difference between TEM and PFDE(P>0.05).Conclusion Tianlongjie can effectively reduce bleomycin-induced alveolitis and fibrosis,early intervention has more significance.The mechanism may related to the inhibition of the expression of TGF-β_(1) in lung tissue,which is an important factor affects the occurrence and development of PF.