姜黄素拮抗瘦素损伤小鼠足细胞的实验研究

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.

Curcumin Antagonizes the Effect of Leptin on Mice Podocytes Injury

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.