肺痹方干预肺纤维化模型小鼠细胞外基质转化的机制  

Interventional mechanism of Feibi prescription on extracellular matrix transformation in a mouse model of pulmonary fibrosis

作　　者：程雪[1] 方泓[2] 张运克[1] 吴银根[2] Cheng Xue;Fang Hong;Zhang Yunke;Wu Yingen(Henan University of Chinese Medicine,Zhengzhou 450046,Henan Province,China;Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]河南中医药大学,河南省郑州市450046 [2]上海中医药大学附属龙华医院,上海市200032

出　　处：《中国组织工程研究》2020年第31期5038-5043,共6页Chinese Journal of Tissue Engineering Research

基　　金：河南省高等学校重点科研项目(20B360015),项目负责人:程雪;第二期名师传承工程吴银根名师研究室(教学009),项目负责人:方泓。

摘　　要：背景:肺组织损伤修复过程中,细胞外基质成分首先填补损伤肺组织,当其过度沉积时可形成肺间质纤维化。目的:探讨肺痹方对博莱霉素致肺纤维化小鼠细胞外基质转化的干预作用。方法:将60只C57BL/6雄性小鼠随机分为空白对照组、模型组、吡非尼酮组、肺痹方低剂量组、肺痹方中剂量组、肺痹方高剂量组,每组10只。除空白对照组外,其余5组腹腔注射博莱霉素[7.5 mg/(kg·d)]建立肺纤维化模型,连续注射10 d。造模后第1天各药物组灌胃给药[51.43 mg/(kg·d)]吡非尼酮,[6.43,12.86,25.72 mg/(kg·d)肺痹方],连续给药28 d。用药28 d后取肺组织,采用苏木精-伊红染色和Masson染色观察小鼠肺组织的形态学变化,ELISA法检测血清中转化生长因子β1、肿瘤坏死因子α水平,Western blot法检测肺组织中α-平滑肌肌动蛋白、Ⅰ型胶原、Ⅲ型胶原的表达。结果与结论:①与空白对照组相比,各组小鼠血清转化生长因子β1水平显著升高(P<0.01);与模型组相比,吡非尼酮组、肺痹方中剂量组、肺痹方高剂量组血清转化生长因子β1水平显著降低(P<0.05,P<0.01),肺痹方高剂量组降低最明显;②与模型组相比,用药各组血清肿瘤坏死因子α水平明显降低(P<0.01),用药各组之间两两比较差异无显著性意义(P>0.05);③与空白对照组相比,各组α-平滑肌肌动蛋白表达均不同程度增高(P<0.05,P<0.01);肺痹方高剂量组α-平滑肌肌动蛋白表达低于肺痹方低、中剂量组,差异有显著性意义(P<0.05);④与空白对照组相比,模型组、肺痹方低剂量组Ⅰ型胶原表达增高(P<0.05,P<0.01);与模型组相比,肺痹方中、高剂量组Ⅰ型胶原表达降低(P<0.05);⑤与空白对照组相比,各组Ⅲ型胶原表达均不同程度增高(P<0.05,P<0.01);肺痹方中、高剂量组以及吡非尼酮组Ⅲ型胶原表达明显低于肺痹方低剂量组(P<0.05,P<0.01);⑥结果表明,肺痹方可以减轻肺纤维化,抑制博来霉BACKGROUND: In the process of repair, extracellular matrix components first fill up the damaged lung tissue. However, excessive deposition of extracellular matrixes can develop pulmonary interstitial fibrosis. OBJECTIVE: To investigate the effect of Feibi prescription on extracellular matrix transformation in mice with pulmonary fibrosis induced by bleomycin. METHODS: Sixty male C57BL/6 mice were randomly divided into blank control group, model group, pirfenidone group, and three Feibi prescription groups(low-, middle-, and high-dose groups). There were 10 mice in each group. Except for the blank control group, the other five groups were intraperitoneally injected with bleomycin(7.5 mg/kg per day) for 10 continuous days to establish the model of pulmonary fibrosis. On day 1 after modeling, the mice in corresponding drug groups were intragastrically administered with pirfenidone(51.43 mg/kg per day) and Feibi prescription(6.43, 12.86, 25.72 mg/kg per day). Drug administration lasted for 28 days. Then, morphological changes of lung tissue in mice were observed by hematoxylin-eosin staining and Masson staining. The contents of transforming growth factor-β1 and tumor necrosis factor-α in the serum were detected by ELISA, and the expression of a-smooth muscle actin, collagen type I and collagen type Ⅲ in the lung tissue was detected by western blot assay. RESULTS AND CONCLUSION: The serum levels of transforming growth factor-β1 were significantly increased in the other groups compared with the blank control group(P < 0.01), but the levels were significantly lower in the pirfenidone, middle-dose and high-dose Feibi prescription groups than in the model group(P < 0.05, P < 0.01), and the most decline was in the high-dose Feibi prescription group. Compared with the model group, the expression of tumor necrosis factor-α was significantly reduced in the four drug groups(P < 0.01), but there was no significant difference between the drug groups(P > 0.05). Compared with the blank control group, the expression of α-smo

关 键 词：肺痹方 肺纤维化 细胞外基质 博莱霉素 C57BL/6小鼠 

分 类 号：R453[医药卫生—治疗学] R318[医药卫生—临床医学]