睾酮通过抑制Rab13表达影响支持细胞屏障通透性

发布时间：2018-03-16 07:45

本文选题：血睾屏障　切入点：支持细胞　出处：《生命科学研究》2014年06期 　论文类型：期刊论文

【摘要】：探讨Rab13 GTPase对血睾屏障(blood testis barrier,BTB)通透性的调节机制。体外分离培养20 d龄大鼠睾丸支持细胞,睾酮处理后测定跨上皮电阻(transepithelial electrical resistance,TER),FITC-鬼笔环肽染色分析F-actin分布,并采用Western印迹检测Rab13在睾酮处理前后的表达变化;si RNA干扰培养支持细胞中Rab13的表达,Western印迹检测沉默效果并检测连接蛋白表达变化;TER测定Rab13沉默前后上皮屏障功能改变,F-actin染色分析Rab13沉默对肌动蛋白丝分布的影响。结果显示,睾酮处理可使体外支持细胞屏障TER值升高,F-actin在膜下聚集增强,且Rab13在睾酮处理后表达量明显降低;Rab13 si RNA处理不影响连接蛋白质的表达,但可引起F-actin在细胞连接处的分布增强,导致体外BTB屏障功能增强。表明Rab13可通过调节F-actin的排布影响体外BTB通透性。
[Abstract]:To investigate the regulatory mechanism of Rab13 GTPase on the permeability of blood testis barrier barrier (BBB), testicular Sertoli cells of 20 day old rats were isolated and cultured in vitro. After testosterone treatment, the distribution of F-actin was determined by transdermal resistor transepithelial electrical resistance assay. Western blot was used to detect the expression of Rab13 before and after testosterone treatment. Western blot was used to detect the expression of Rab13 in cultured Sertoli cells with si RNA interference. Western blot was used to detect the silencing effect and the change of connexin expression. Ter was used to determine the epithelial barrier function before and after Rab13 silencing. The effect of Rab13 silencing on the distribution of actin filaments was analyzed by changing F-actin staining. Testosterone treatment increased the TER value of Sertoli cell barrier in vitro and increased the concentration of F-actin in the membrane, and the expression of Rab13 decreased significantly after testosterone treatment, and the expression of ligation protein was not affected by RNA treatment, but the distribution of F-actin was enhanced at the junction of cells. It was suggested that Rab13 could influence the permeability of BTB by regulating the arrangement of F-actin.
【作者单位】：中国医科大学基础医学院生物化学与分子生物学教研室;
【基金】：国家自然科学基金资助项目(81100462;81370756)
【分类号】：R335

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