肾康注射液对白蛋白诱导活化的肾小管上皮细胞表型转化的实验研究

【作者】 何玉华；

【导师】 叶传蕙；

【作者基本信息】 成都中医药大学 ， 中西医结合临床， 2005， 博士

【摘要】 目的:观察中药复方制剂肾康注射液对白蛋白诱导体外培养的肾小管上皮细胞表型转化的影响,探讨肾康注射液对肾间质纤维化的防治作用及机制。 方法:采用洛汀新(Lotensin)为阳性对照药,以体外培养的肾小管上皮细胞株作为研究对象,分为正常组、白蛋白组、洛汀新组和不同浓度肾康注射液治疗组。应用MTT比色法测定不同浓度SKI对TEC细胞增殖的作用;免疫组织化学法检测TEC细胞角蛋白(CK)、波形蛋白(vimentin)和α-SMA的表达;原位杂交的方法观察TEC分泌Ⅰ型胶原的变化;Western blot检测不同浓度SKI对白蛋白诱导的TEC NF-κ Bp65活性的影响。 结果:在白蛋白诱导下,培养的TEC出现增殖,不同浓度的SKI对TEC增殖具有抑制作用,以SKI高浓度为显著。免疫组化染色显示,TEC在白蛋白作用下,CK减弱,vimentin、α-SMA表达增加,原位杂交显示Ⅰ型胶原基因表达增加。SKI能明显抑制TEC的表型转化及Ⅰ型胶原基因表达。Western blot显示NF-κ Bp65表达增加,SKI能显著抑制白蛋白培养的TEC NF-κ Bp65的活性。 结论:白蛋白培养TEC细胞明显增殖,发生了上皮向间叶的表型转化。SKI能抑制细胞增殖和表型转化,使CK表达增加,vimentin、α-SMA和Ⅰ型胶原的的表达减弱。SKI可使NF-κB的活性下降,这可能是SKI抑制TEC表型转化的重要机理之一。更多还原

【Abstract】 Objective: To investigate the effects and mechanisms of Shenkang Injection, a compound TCM preparation, on transdifferentiation of Renal Tubular Epithelial cells induced by BSA in vitro and explore the effects of Shenkang Injection to prevent and treat renal interstitial fibrosis.Methods: Using Lotensin as positive control and TEC as study object. TEC was divided into 6 groups, control, BSA-treated group, Lotensin-treated group, Shenkang Injection-treated groups with different dose. The cell proliferation was observed by MTT assay, Immunohistochemistry stain was used to examine the expression of cytokeratin,vimentin and α -SMA .in situ hybridization was used to examined the expression of collagen I gene, and Western blot was applied to further detect the process of NF-κBp65 protein expression.Results:BSA increased cellular proliferation,Various concentrations of Shenkang Injection showed an obvious inhibition effect to the cellular proliferation, and the effect is better than Lotensin especially in high更多还原