【摘要】 目的:探讨瘦素对小鼠足细胞的损伤效应,及姜黄素拮抗这一效应的可能。方法:小鼠足细胞分为四组,分别为空白对照组,瘦素刺激组,姜黄素加瘦素组,及姜黄素对照组。mRNA检测采用实时定量PCR方法,蛋白检测采用免疫印迹法。结果:瘦素(15 ng/ml)能显著抑制足细胞nephrin,podocin,podoplanin,podocalyxin表达,与对照组比较,mRNA表达的抑制率分别为31%、28%、33%及29%;蛋白质表达的抑制率分别为26%、30%、24%及32%,P均<0.05。加入姜黄素(5μmol/L)后,上述足细胞标志蛋白均被上调,与瘦素组比较,mRNA表达分别上调1.25、1.15、1.34及1.20倍;蛋白质表达分别上调1.15、1.31、1.14及1.32,P均<0.05。瘦素(15 ng/ml)能显著活化Wnt/β-catenin信号通路,与对照组比较,Wnt1、Wnt2b和Wnt6及其下游蛋白β-catenin的mRNA表达分别上调1.54、1.29、1.52及1.25倍,P均<0.05。β-catenin的蛋白磷酸化水平抑制率为17%,P<0.05。加入姜黄素(5μmol/L)后,上述信号通路分子均被抑制,与瘦素组比较,Wnt1、Wnt2b和Wnt6及β-catenin的mRNA表达抑制率分别为12%、12%、22%及10%,P<均0.05。磷酸化的β-catenin上调1.13倍。结论:瘦素能通过Wnt/β-catenin信号通路损伤足细胞,而姜黄素能逆转这一反应。更多还原

【Abstract】 Objective: To investigate the effect of leptin on mice podocytes injury,and curcumin might antagonize this effect. Methods: Podocyte were incubated with medium alone,medium with curcumin( 5 μmol / L),medium with leptin( 15 ng / ml) or medium with curcumin( 5 μmol / L) and leptin( 15 ng / ml),respectively. mRNA expression was detected by real-time quantitative PCR,protein expression was detected by Western blotting. Results: The mRNA and protein expression of nephrin,podocin,podoplanin,podocalyxin were significantly down-regulated by leptin( 15 ng / ml). Compared with the control group,The inhibition rates of nephrin,podocin,podoplanin,podocalyxin mRNA in the leptin group were 31%,28%,33% and 29%,respectively( P < 0. 05),and the inhibition rates of their protein expression were 26%,30%,24% and 32%,respectively( P < 0. 05). Curcumin significantly up-regulated the down-regulation of nephrin,podocin,podoplanin,podocalyxin induced by leptin. Compared with the leptin group,the mRNA expression of nephrin,podocin,podoplanin,podocalyxin were up-regulated to 1. 25,1. 15,1. 34 and 1. 20 times,respectively( P < 0. 05),their protein expression were up-regulated to 1. 15,1. 31,1. 14 and 1. 32 times,respectively( P <0. 05). Leptin( 15 ng/ml) can significantly activate Wnt/β-catenin signaling pathway. Compared with the control group,mRNA expression of Wnt1,Wnt2b,Wnt6 and β-catenin were up-regulated 1. 54,1. 29,1. 52 and 1. 25 times,respectively( P < 0. 05). The inhibition rates of phospho-β-catenin in the leptin group was 17%( P < 0. 05). Curcumin significantly inhibited the activation of Wnt / β-catenin signaling pathway induced by leptin. Compared with the leptin group,the inhibition rates of Wnt1, Wnt2b and Wnt6 and β-catenin mRNA in the curcumin and leptin costimulated group were 12%,12%,22% and 10%,respectively( P < 0. 05). The phosphor-β-catenin in the curcumin and leptin costimulated group were up-regulated 1. 13 times( P < 0. 05). Conclusion: Leptin can cause podocyte injury through Wnt / β-catenin signaling pathway,curcumin can reverse this effect induced by leptin.更多还原