TRB3-在非诺贝特抑制高糖诱导的肾小球系膜细胞增殖中的作-课件.pdfVIP

J South Med Univ, 2013, 33(3): 391-396 doi 10.3969/j.issn. 1673-4254.2013.03.17 ·391 · 基础研究 TRBTRB33 在非诺贝特抑制高糖诱导的肾小球系膜细胞增殖中的作在非诺贝特抑制高糖诱导的肾小球系膜细胞增殖中的作 用及其机用及其机制制 宋青青，孙 嘉，王 娇，杨 锐，陈容平，张 振，罗海钊，蔡德鸿 南方医科大学珠江医院内分泌科，广东 广州 510282 摘要：目的探讨TRB3 在非诺贝特抑制高糖诱导下肾小球系膜细胞增殖中的作用及作用其机制。方法 将细胞分为正常对照 组（N ，5.5 mmol/L 葡萄糖）、高糖组（H ，25 mmom/L 葡萄糖）、高糖+不同浓度的非诺贝特组（FN ，10、50、100 μmol/L）。采用 CCK-8法检测细胞增殖率，Hoechst33258染色观察细胞凋亡形态学改变，流式细胞术检测细胞周期改变，免疫细胞化学染色观 察TRB3表达，Western Blot 检测细胞中TRB3、P-AKT 蛋白的表达。结果高糖能够诱导肾小球系膜细胞增殖（P0.001）；非诺贝 特能够抑制肾小球系膜细胞增殖（P0.001），且具有浓度依赖性。非诺贝特干预后细胞出现凋亡形态学改变；非诺贝特可以使 系膜细胞发生G1/S 期阻滞；正常组、高糖组胞浆有少量TRB3 蛋白表达，但高糖不能促进TRB3 表达增多，随着非诺贝特浓度的 增加TRB3 表达量增多，P-AKT 表达逐渐降低。结论非诺贝特可以促进TRB3 的表达；TRB3 可能通过抑制AKt 的磷酸化使肾 小球系膜细胞发生G1/S 期阻滞从而抑制其增殖。 关键词：非诺贝特；肾小球系膜细胞；TRB3 ；Akt ；细胞周期 Role of TRB3 in the inhibitory effect of fenofibrate against high glucose-induced proliferation of glomerular mesangial cells SONG Qingqing, SUN Jia, WANG Jiao, YANG Rui, CHEN Rongping, ZHANG Zhen, LUO Haizhao, CAI Dehong Department of endocrinology, Zhujiang Hospital , Southern Medical University, Guang Zhou, 510282, China Abstract: Objective To investigate the role of TRB3 in the inhibitory effect of fenofibrate against the proliferation of glomerular mesangial cell induced by high glucose. Methods Rat glomerular mesangial cells (MCs) were cultured in the presence of 5.5 mmol/L glucose (normal control), 25 mmol/L glucose (high glucose group), or high glucose along with 10, 50, or 100 μmol/L fenofibrate. Cell counting kit-8 (CCK-8) assay was used to evaluate cell proliferation, and Hoechst 33258 staining was employed to determine chromatin distribution in the MCs. Flow cytometry was performed to analyze the cell cycle changes in different groups. The expressions of TRB3 and P-AKT in different groups were detected using immunocytochemistry and Western blotting. Results H