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  

Paeonol up-regulates CKIP-1 to resist high glucose-induced fibrosis in glomerular mesangial cells

作　　者：张蕾 邹叶子 公文艳 陈志泉 黄河清[1] ZHANG Lei;ZOU Ye-zi;GONG Wen-yan;CHEN Zhi-quan;HUANG He-qing(Lab of Pharmacology and Toxicology,School of Pharmaceutical Science of Sun Yat-Sen University,Guangzhou 510006,China)

机构地区：[1]中山大学药学院药理与毒理学实验室,广东广州510006

出　　处：《中国药理学通报》2018年第5期645-650,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81573477;81770816);广东省自然科学基金重点项目(No 2017A030311036)

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H_2O_2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 inhibit 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 expressions of CKIP-1,Nrf2,FN and ICAM-1 were evaluated 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 technique 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 antioxidant enzymes,i.e.HO-1 and SOD1,were also up-regulated by paeonol,which was accompanied by reductions of superoxide and H 2O 2 levels.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 H 2O 2 levels.Furthermore,si-CKIP-1 reversed the down-regulated levels of FN and ICAM-1 induced by paeonol.Conclusion Paeonol inhibits the expressions of FN and ICAM-1 in HG-treated GMCs possibly by up-regulating CKIP-1 and activating the Nrf2 signaling pathway.

关 键 词：丹皮酚 高糖 CKIP-1 Nrf2 纤维连接蛋白 肾小球系膜细胞 肾脏纤维化 

分 类 号：R-332[医药卫生] R284.1