丹皮酚激活CKIP-1对高糖诱导的肾小球系膜细胞纤维化的影响

摘要

目的 观察丹皮酚通过激活CKIP-1,活化Nrf2抗氧化应激通路,进而抵抗高糖诱导的肾小球系膜细胞(glomerular mesangial cells,GMCs)中纤维连接蛋白(fibronectin,FN)和细胞间黏附分子1(intercellular cell adhension molecule-1, ICAM-1)表达增加的作用.方法 采用高糖刺激的 GMCs体外模型,观察丹皮酚对模型细胞FN、ICAM-1等炎性纤维化成分表达及CKIP-1、Nrf2抗氧化通路的影响;采用小分子RNA干扰GMCs CKIP-1的蛋白表达,免疫印迹法检测GMCs内Nrf2、HO-1、SOD1蛋白表达,DHE荧光探针技术检测细胞内超氧化物含量.结果 在高糖诱导的GMCs,丹皮酚能上调CKIP-1蛋白的表达,增加Nrf2的水平,以及Nrf2信号通路下游靶因子HO-1、SOD1的表达,有效减少胞内超氧化物和H2O2含量,最终逆转 FN、ICAM-1表达的增加.在干扰CKIP-1表达后,丹皮酚升高Nrf2及HO-1、SOD1的作用明显减弱,不能有效减少胞内活性氧水平,最终不能下调 FN、ICAM-1的蛋白表达.结论 激活CKIP-1,活化Nrf2信号通路,可能是丹皮酚抑制高糖引起的FN、ICAM-1上调的分子机制之一.%Aim To observe whether paeonol can in-hibit fibronectin (FN) and intercellular cell adhension molecule-1 (ICAM-1) expressions in high glucose (HG)-induced glomerular mesangial cells(GMCs) via up-regulating CKIP-1 and activating the Nrf2 signaling pathway. Methods The effects of paeonol on the ex-pressions of CKIP-1,Nrf2,FN and ICAM-1 were eval-uated in GMCs treated with HG. Small interfering RNA was used to deplete CKIP-1 protein expression, and Western bolt was used to detect the expressions of Nrf2, HO-1 and SOD1. DHE fluorescent probe tech-nique was used to determine intracellular superoxide level. Results The protein levels of CKIP-1 and Nrf2 were elevated by paeonol in HG-treated GMCs. In the meanwhile,the expressions of Nrf2 downstream antiox-idant enzymes, i.e. HO-1 and SOD1, were also up-regulated by paeonol, which was accompanied by re-ductions of superoxide and H2O2levels. Importantly, paeonol reversed the excessive accumulation of FN and ICAM-1 in HG-induced GMCs. si-CKIP-1 decreased the up-regulation of Nrf2,HO-1 and SOD1 expressions during paeonol treatment, which was accompanied by increased superoxide and H2O2levels. Furthermore, si-CKIP-1 reversed the down-regulated levels of FN and ICAM-1 induced by paeonol. Conclusion Pae-onol inhibits the expressions of FN and ICAM-1 in HG-treated GMCs possibly by up-regulating CKIP-1 and activating the Nrf2 signaling pathway.