激活素A及卵泡抑素对大鼠肾间质成纤维细胞分泌羟脯氨酸及Ⅵ型胶原的影响.docVIP

激活素A及卵泡抑素对大鼠肾间质成纤维 细胞分泌羟脯氨酸及Ⅵ型胶原的影响 第三军医大学新桥医院肾内科，全军肾脏病中心，重庆市肾病研究所（400037） 李芙蓉，，袁发焕*摘　要 目的： 。方法：取SD大鼠肾间质成纤维细胞进行体外培养，分别向培养液中加入激活素A或激活素A+卵泡抑素卵泡抑素，用酶联免疫吸附法检测培养液上清中羟脯氨酸Ⅳ型胶原的含量。结果： ACT-A浓度越高，培养液中浓度也越高，单纯加入FS对大鼠肾间质成纤维细胞分泌浓度没有影响，但以剂量依赖方式抑制ACT-A促大鼠肾间质成纤维细胞分泌的作用。结论：激活素A可能在肾间质损害的发生发展中起重要作用补充外源性的FS有可能为治疗肾间质纤维化新的思路　 关键词：激活素A泡抑素；成纤维细胞Ⅳ型胶原羟脯氨酸Effects of activin and follistain on hydroxyproline and type Ⅳ collagen of cultured rat renal interstitial fibroblasts LI Fu-rong,Yang Cheng, Zhang Ying,Yuan Fahuan. Department of Nephrology, Xinqiao Hospital, Third Military Medical University , Chongqing 400037 , China Objective: To further study the effects of activin A (ACT-A) and follistatin (FS) on cultured rat renal interstitial fibroblasts. Methods: Fibroblasts separated from SD rat kidney were cultured in different medium contained ACT-A or ACT-A and FS or FS in different concentration. Levels of type Ⅳ collagen and hydroxyproline（HYP） in the supernatants of cultured fibroblasts were measured by ELISA assay. Results: ACT-A stimulated expression of type Ⅳ collagen and hydroxyproline in cultured rat renal interstitial fibroblasts in dose-dependent manner in the range from 0.3 ng/ml to 100 ng/ml. FS inhibited the effects of ACT-A on Collagen Ⅳ and hydroxyproline in supernatants of cultured fibroblasts in dose-dependent manner. Conclusion：ACT-A might renal fibrosis by affecting renal interstitial fibroblasts. FS could suppress the effects of ACT-A on the cultured renal fibroblasts. Key words: activin A; follistatin ; fibroblasts; type Ⅳ collagen；hydroxyproline 激活素(Activin，ACT) 是近年来发现的一组多功能的细胞因子, 1986 年首次从牛的卵泡液中分离、纯化，根据其促进卵泡刺激素(FSH) 分泌及合成的生物学活性而命名为激活素。ACT是由两条β亚单位借二硫键构成的二聚体蛋白, 属于转化生长因子β(transforming growth factor β,TGF-β) 基因超家族成员之一。根据其亚单位的结合形式不同, 又分别形成激活素AB (βA–βB)、激活素B (βB–βB)和激活素A (βA–βA ) 3种分子结构。最近βC , βD和βE 已经被发现调节ACT活动最重要的蛋白是ACT结合蛋白－卵泡抑素(follistatin，FS)。FS以高亲和力结合到ACT并阻止其生物学效应[]。、活化[2]，可能在肾间质的纤维化中起重要作用，FS抑制了ACT-A的这一生物学特性。关于ACT-A及FS在肾间质纤维化中的作用还不是很清楚。本研究拟通过原代分离大鼠肾间质成纤维细胞（rat renal