【作者】 吴娟娟；

【导师】 凌春华；

【作者基本信息】 苏州大学 ， 呼吸内科学， 2012， 硕士

【摘要】 目的：探讨复方三七饮（compound panax notoginsenoside granules，CPNG）对博莱霉素（bleomycin，BLM）致大鼠肺纤维化的干预作用及其机制。方法：雄性SD大鼠60只，随机分为6组：正常组、模型组、醋酸泼尼松组、CPNG大、中、小剂量组[CPNG（H、M、L）]，每组10只。经气管内注入BLM（5mg/kg）制备大鼠肺纤维化模型。造模第二天，治疗组分别灌服醋酸泼尼松(3.33mg·kg^-1·d^-1)、CPNG大、中、小剂量(100、50、25mg·kg^-1·d^-1)，正常组和模型组大鼠灌服等体积的蒸馏水，每天1次。28天后处死大鼠，计算肺系数；碱水法测定肺组织中羟脯氨酸（HYP）含量；HE染色和MASSON染色进行肺组织病理学观察；血清和肺组织中的丙二醛（MDA）含量、还原性谷胱甘肽（GSH）含量、超氧化物歧化酶（SOD）活力和总抗氧化（T-AOC）能力，分别用硫代巴比妥酸（TBA）法、化学比色法、黄嘌呤氧化酶法和化学比色法测定。免疫组织化学染色检测转化生长因子-β1（TGF-β1）、Smad2、Smad3、平滑肌肌动蛋白-α（α-SMA）和胶原蛋白^-1（Col-Ⅰ）蛋白的表达水平；Western-blot检测TGF-β1、Smad2、Smad7蛋白的表达水平。结果：与模型组比较，CPNG各剂量组大鼠肺系数降低（P<0.05，P<0.01），肺组织中HYP降低（P<0.01）；血清与肺组织匀浆中的丙二醛（MDA）含量降低（P<0.001），血清和肺组织匀浆中谷胱甘肽（GSH）含量、超氧化物歧化酶（SOD）活性、总抗氧化能力（T-AOC）升高（P<0.05，P<0.01，P<0.001）。肺组织的病理组织学HE和Masson染色显示CPNG能明显减轻大鼠肺泡炎和肺纤维化程度（P<0.01，P<0.01）。免疫组织化学和Western-blot结果显示：与模型组大鼠比较，CPNG各剂量组TGF-β1、Smad2、Smad3、α-SMA、Col-Ⅰ蛋白的表达水平降低（P<0.05，P<0.01，P<0.01），Smad7蛋白表达水平升高（P<0.01）。结论：复方三七饮对博莱霉素致大鼠肺纤维化具有一定的防治作用，其机制可能与抑制氧化应激，调节TGF-β1/Smads信号通路、抑制胶原蛋白的形成和沉积有关。更多还原

【Abstract】 Objective：To explore the effects of compound panax notoginsenoside granules（CPNG） onbleomycin-induced pulmonary fibrosis in rats and its mechanisms.Methods：Sixty male Sprague-Dawley rats were randomly divided into6groups, includingcontrol normal group，model group, prednisolone group(3.33mg·kg^-1·d^-1)and CPNGhigh-dose, middle-dose and low-dose group(100、50、25mg·kg^-1·d^-1)，10Sprague-Dawleyrats in each group. Except the control normal group, pulmonary fibrosis models werereproduced by intratracheal injection of bleomycin（5mg/kg）. On the second day ofinjection of bleomycin, the rats in each treatment group were intragastric administratedprednisolone and corresponding dose of CPNG daily; control normal group and modelgroup were intragastric administrated the equal volume distilled water daily. After28days,all rats were executed to be observed the pulmonary index. Hydroxyproline （HYP） in lungtissue was assessed. Fixed parts of the lung tissue was obtained for HE and Masson stainfor histopathological examination. Malondialdehyde （MDA）, glutathion （GSH）, superoxidedismutase（SOD） and total antioxidant capacity （T-AOC） in blood and lung tissue wererespectively assessed by means of Thibabituric Acid（TBA）, chemical colorimetry,xanthine oxidase and chemical colorimetry. The expressions of transforming growth factorβ1（TGF-β1）, Smad2, Smad3, α-smooth muscle actin （α-SMA） and collagen I （Col-I）protein were analyzed by immunohistochemistry. The expressions of TGF-β1, Smad2,Smad7protein were analyzed by western-blotting. Results:Compared with model group, the pulmonary index and lung tissue HYP weredecreased （P<0.05，P<0.01）, MDA level in blood and lung tissue was also decreased（P<0.001）, but GSH, SOD, T-AOC level in blood and lung tissue were significantlyincreased （P<0.05，P<0.01，P<0.001）in all of CPNG groups. The histopathology of lungtissue with HE and Masson staining demonstrated that CPNG dramatically decreased thelung inflammation and lung fibrosis in rats（P<0.01，P<0.01）. Munohistochemistry andwestern-blotting demonstrated that CPNG could inhibite the expressions of TGF-β1,Smad2, Smad3, α-SMA and Col-I protein（P<0.05，P<0.01，P<0.01）, and increase theexpression of Smad7protein in lung tissue of rats（P<0.01）.Conclusion:CPNG has a therapeutic effect on bleomycin-induced pulmonary fibrosis in rats,which maybe relate to antioxidative damage, intervention TGF-β1/Smads signalingpathway, and restraining the formation and deposition of collagen.更多还原