Wnt5a/TRPC6在姜黄素拮抗肥胖相关肾小球病足细胞损伤中的机制探讨  

The Role of Wnt5a/TRPC6/Ca2+ in Curcumin Antagonizing Obesity-Related Glomerulopathy Podocyte Injury

作　　者：刘雪姣[1] 赵京 王国勤[1] 芮宏亮[1] 杨敏[1] 王艳艳[1] 谌贻璞[1] 程虹[1] LIU Xuejiao;ZHAO Jing;WANG Guoqin(Division of Nephrology,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029)

机构地区：[1]首都医科大学附属北京安贞医院肾内科

出　　处：《中国中西医结合肾病杂志》2019年第12期1044-1047,共4页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.81641143、81573745);北京市自然科学基金资助项目(No.7172066)

摘　　要：目的:探讨Wnt5a/TRPC6/Ca 2+在姜黄素拮抗肥胖相关肾小球病(ORG)足细胞损伤中的作用及机制。方法:小鼠足细胞分为4组:对照组、瘦素刺激组、Wnt5a刺激组和姜黄素干预组。mRNA检测采用实时定量PCR方法,蛋白检测采用免疫印迹法。应用Fura 2-AM检测细胞内钙离子浓度。结果:(1)与对照组相比,瘦素刺激12 h后,Wnt5a、TRPC6的mRNA和蛋白的表达均显著增高,podocin、nephrin的mRNA和蛋白的表达均显著降低;与瘦素刺激组相比,姜黄素干预组中Wnt5a、TRPC6的mRNA和蛋白的表达均显著降低,podocin、nephrin的mRNA和蛋白的表达显著增高。(2)与对照组相比,Wnt5a刺激12 h后,TRPC6的mRNA和蛋白的表达均显著增高,podocin、nephrin的mRNA和蛋白的表达均显著降低。(3)与对照组相比,加入瘦素刺激后,从7 min开始足细胞内钙离子水平明显增多,至15 min时逐渐趋于平稳;而姜黄素干预组中,加入瘦素刺激后监测20 min,足细胞内钙离子浓度始终趋于平稳。结论:Wnt5a/TRPC6/Ca 2+通路活化是ORG足细胞损伤的重要机制之一,肾脏足细胞Wnt5a活化可以刺激TRPC6表达上调,致足细胞损伤;姜黄素可能通过抑制Wnt5a/TRPC6/Ca 2+通路的活化来拮抗ORG足细胞损伤。Objective:To investigate the effect of Wnt5a/TRPC6/Ca 2+on ORG podocytes injury,and curcumin might antagonize this effect.Methods:Podocyte were incubated with medium alone,medium with leptin,medium with Wnt5a or medium with curcumin and leptin,respectively.The expression of mRNA was detected by real-time quantitative PCR,protein expression was detected by Western blotting.Calcium imaging experiments performed with the fluorescent Ca 2+indicator Fura 2-AM.Results:(1)Compared with the control group,the expression of Wnt5a and TRPC6 were significantly up-regulated by leptin,and the expression of nephrin and podocin were significantly down-regulated by leptin.Compared with the leptin group,the expression of Wnt5a and TRPC6 were significantly down-regulated by curcumin,and the expression of nephrin and podocin were significantly up-regulated by curcumin.(2)Compared with the control group,the expression of TRPC6 was significantly up-regulated by Wnt5a,and the expression of nephrin and podocin were significantly down-regulated by Wnt5a.(3)Comparing with the control group,the level of calcium in podocytes increased significantly from 7 minutes to 15 minutes after leptin stimulation.In curcumin protection group,the concentration of calcium in podocytes remained always stable.Conclusion:Activation of Wnt5a/TRPC6/Ca 2+pathway is one of the important mechanisms of ORG podocyte injury.Activation of Wnt5a in renal podocytes can cause up-regulation of TRPC6 expression,resulting in podocyte injury.Curcumin may alleviate ORG podocyte injury by inhibiting the activation of Wnt5a/TRPC6/Ca 2+pathway.

关 键 词：姜黄素 瘦素 肥胖相关性肾小球病 WNT5A TRPC6 Ca2+ 通道 

分 类 号：R285.5[医药卫生—中药学]