益气活血方对肾间质纤维化大鼠肾脏TGF-β/smad信号通路及结缔组织生长因子的影响被引量：14

Effects of Chinese herbal medicine Yiqi Huoxue Formula on TGF-β/smad signal transduction pathway and connective tissue growth factor in rats with renal interstitial fibrosis

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摘要目的:观察益气活血方对肾间质纤维化大鼠肾脏TGF-β/smad信号通路及结缔组织生长因子(connectivetissue growth factor,CTGF)的影响。方法:健康雄性Wistar大鼠30只诱导建立单侧输尿管梗阻(unilateral ureteral obstruction,UUO)致肾小管间质纤维化模型,模型成功后随机分为模型组,益气活血方大、中、小剂量组和西药组,另取健康雄性Wistar大鼠6只设为假手术组。3周后处死大鼠,称取体质量;取血清检测肌酐(creatinine,Cr)、尿素氮(bloodurea nitrogen,BUN)水平;取肾脏组织采用实时荧光定量逆转录聚合酶链反应法和免疫印迹法检测各组转化生长因子β(transforming growth factor-β,TGF-β)、smad2、smad7、CTGF mRNA和蛋白表达水平;苏木精-伊红染色和Masson染色检测各组肾间质纤维化指数。结果:模型组大鼠血清Cr和BUN值较假手术组偏高;与模型组比较,大剂量益气活血方可以降低Cr和BUN值,但差异无统计学意义。苏木精-伊红染色和Masson染色显示模型组肾小管上皮细胞萎缩,大部分肾小管管腔扩张,间质纤维组织增生,大量炎性细胞浸润,肾小球数目明显减少,胶原成分显著增加。益气活血方大剂量组、中剂量组和西药福辛普利钠组大鼠肾间质炎性细胞浸润、纤维组织增生、肾小管扩张情况较模型组明显减轻,胶原成分明显减少。与假手术组比较,模型组大鼠肾脏组织中TGF-β、smad2和CTGFmRNA以及蛋白表达水平明显升高(P<0.05),smad7的表达水平则明显降低(P<0.05)。与模型组比较,益气活血方大剂量组和中剂量组大鼠TGF-β、smad2、CTGF mRNA和蛋白表达明显下降(P<0.01,P<0.05),smad7表达明显升高(P<0.01,P<0.05)。结论:UUO大鼠肾组织中CTGF mRNA的表达可能通过TGF-β/smad信号通道进行调节;益气活血方可以通过下调TGF-β、smad2和上调smad7的表达来抑制CTGF表达,进而遏制肾纤维化的进展。 Objective： To observe the effects of Yiqi Huoxue Formula （YQHXF）, a compound Chinese herbal medicine, on transforming growth factor-β（TGF-β/smad signal transduction pathway and connective tissue growth factor （CTGF） in rats with renal interstitial fibrosis Methods： Unilateral ureteral obstruction （UUO） rat model was established and the rats were randomly divided into 5 groups： untreated group, high-, medium-, and low-dose YQHXF groups and fosinopril sodium group. Another group with sham operation was set as control. All rats were administered with corresponding drugs for 3 weeks. After the last administration, each rat was sacrificed and weighed and the serum was separated for creatinine （Cr） and blood urea nitrogen （BUN） detection. Kidneys of the rats were taken out, and mRNA and protein expressions of TGF-β, smad2, smad7 and CTGF were measured with real-time fluorescent quantitative reverse transcription-polymerase chain reaction and Western blotting respectively; fibrosis of the kidney tissue was observed with hematoxylin-eosin （HE） staining and Masson trichrome staining. Results： Compared with sham-operation group, Cr and BUN in serum of UUO groups were increased, while high-dose YQHXF treatment decreased the UUO-induced increase of Cr and BUN levels. HE staining and Masson staining results showed that the renal tubular epithelial cells in untreated group got atrophied; lumens of renal tubules expanded; fibroplastic proliferation and inflammatory cell infiltration were observed in renal interstitium; the number of glomerulus decreased and collagen increased significantly compared with shamoperation group. In the high- and medium-dose YQHXF groups and fosinopril sodium group, the histopathological changes of inflammatory cell infiltration, fibroplastic proliferation, expansion of lumens of renal tubules was improved as compared with the untreated group. The mRNA and protein expressions of TGF-β, smad2 and CTGF in untreated group were higher than those in sham-operation group （P〈0. 05）, and the mRNA and protein expressions of smad7 in untreated group were lower than those in the sham-operation group （P〈0. 05）. Compared with untreated group, high- and medium-dose of YQHXF significantly down-regulated the mRNA and protein expressions of TGF-β, smad2 and CTGF （P〈0. 01, P〈0. 05）, and up-regulated the mRNA and protein expressions of smad7 （P〈0. 01, P〈0. 05）. Conclusions： The mRNA expression of CTGF in UUO rats may be regulated by TGF-β/smad signaling transduction pathway. YQHXF might inhibit the expression of CTGF through down-regulation of TGF-β and smad2 and up-regulation of smadT, thus inhibiting the progression of renal interstitial fibrosis.

作者刘咏梅刘瑞华刘文军刘丽吴志奎陈玉英

机构地区中国中医科学院广安门医院分子生物学研究室首都医科大学解剖与组织胚胎学系

出处《中西医结合学报》 CAS 2010年第12期1165-1173,共9页 Journal of Chinese Integrative Medicine

基金国家自然科学基金资助项目(No.30873251) 北京市自然科学基金资助项目(No.7062057)

关键词复方(中药) 转化生长因子β SMAD蛋白质类结缔组织生长因子肾间质纤维化大鼠 compounds （TCD） transforming growth factor-beta smad proteins connective tissue growth factor renal interstitial fibrosis rats

分类号 R285.5 [医药卫生—中药学]

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