地塞米松影响足细胞活动力的相关分子研究  

Molecular mechanism of dexamethasone affecting podocyte motility

作　　者：刘海梅[1] 徐虹[1] 高霞[1] 郑鹢冰[1] 刘俊朝[1] 查锡良[2] 

机构地区：[1]复旦大学附属儿科医院肾脏风湿科,上海201102 [2]复旦大学医学院分子生物学与生物化学系

出　　处：《中华肾脏病杂志》2012年第9期705-709,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金（30971375） 志谢 感谢复旦大学医学院分子生物与生物化学系老师对本研究给予的技术支持;感谢黄文彦教授在实验设计和实施中给予的宝贵意见;感谢苏州儿童医院李晓忠教授在细胞株培养中提供的帮助

摘　　要：目的 通过分析地塞米松对足细胞活动力的影响特点,探讨其对足细胞固有分子α-actinin-4、nephrin的调控能力.方法 体外足细胞根据不同处理分为3组：地塞米松组（地塞米松1μmol/L＋嘌呤霉素50 mg/L）、嘌呤霉素组（嘌呤霉素50 mg/L）和正常对照组.采用划痕和Transwell实验检测各组足细胞活动力的变化.应用两室弥散系统检测各组足细胞单层膜对FITC-BSA的滤过率.使用实时定量PCR和Western印迹法分析各组足细胞α-actinin-4和nephrin的表达变化.结果 足细胞在嘌呤霉素干预后,细胞划痕修复能力及细胞迁移数均显著提高（P＜0.01）.与嘌呤霉素组比较,地塞米松预处理后足细胞再次给予同样剂量、时间的嘌呤霉素刺激后,细胞划痕修复及迁移数均未发生显著升高（P＜0.01）.实时定量PCR和Western印迹结果均提示,地塞米松预处理后足细胞α-actinin-4和nephrin的表达量显著低于嘌呤霉素组（P＜0.05）.结论 地塞米松能够减轻嘌呤霉素诱导的足细胞活动力升高,其作用机制可能与调控α-actinin-4和nephrin的表达有关.Objective To investigate the potential roles of dexamethasone （Dex） in podocyte motility,and to explore the mechanism of modulating α-actinin-4,nephrin.Methods Podocytes were divided into three groups：Dex group [1 μmol/L Dex ＋50 mg/L puromycin aminonucleoside （PAN）],PAN group （50 mg/L） and normal control group.Scrape wound assay and Transwell migration assay were used to detect cell motility.Filtering ratio of podocytes was measured by FITC labeled BSA.Real-time PCR and Western blotting were used to examine the expressions of c-actinin-4 and nephrin.Results From the scrape wound assays,the ability of wound repair in podocytes of PAN group was significantly increased （P〈0.01）,and the number of migrating cells in this group also rose （P〈0.01）.Compared to PAN group,podocytes in Dex group did not enhance the motility after treatment with the same dose PAN （P〈0.01）.Real-time PCR and Western blotting showed that Dex could significantly inhibit the up-regulated expression of α-actinin-4 and nephrin induced by PAN.Conclusions Dex can relieve the enhanced motility induced by PAN.Its mechanism may be associated with the modulation of the expressions of α-actinin-4 and nephrin.

关 键 词：地塞米松 足细胞 辅肌动蛋白 细胞运动 NEPHRIN 

分 类 号：R96[医药卫生—药理学]