Smurf2靶向EYA2对高糖诱导肾小球系膜细胞增殖和纤维化的影响

Effect of Smurf2 Targeting EYA2 on Proliferation and Fibrosis of Glomerular Mesangial Cells Induced by High Glucose

摘要

目的 探究Smurf2通过靶向EYA2影响高糖诱导肾小球系膜细胞增殖和纤维化的分子机制。方法 培养小鼠肾小球系膜细胞（GMCs）,将sh-Smurf2与sh-NC载体转染到高糖诱导的GMCs中,分组为：空白对照组（NG）、高糖处理组（HG）、高糖+sh-NC组（HG+sh-NC）、高糖+sh-Smurf2组（HG+sh-Smurf2）。CCK-8实验和EdU实验检测各组细胞增殖,蛋白印迹法（Western blot）检测各组细胞Fibronectin、Collagen I、α-SMA蛋白的表达;采用CO-IP实验检测Smurf2与EYA2的靶向互作关系,进一步采用sh-EYA2与sh-NC载体转染GMCs中进行回溯实验验证。结果 与NG组相比,HG组细胞增殖显著上升,Fibronectin、Collagen I、α-SMA蛋白表达显著升高;与HG组相比,干扰Smurf2可以抑制细胞增殖,并抑制Fibronectin、Collagen I、α-SMA蛋白的表达。CO-IP结果显示Smurf2与EYA2存在靶向互作关系,且与HG+sh-Smurf2组相比,干扰EYA2可以促进细胞增殖,并促进Fibronectin、Collagen I、α-SMA蛋白的表达。结论 干扰Smurf2可以通过上调EYA2抑制高糖诱导的GMCs过度增殖与纤维化。

Abstract

Objective To explore the molecular mechanism of Smurf2's effect on hyperglycemia-induced mesangial cell proliferation and fibrosis by targeting EYA2. Methods Mouse mesangial cells (GMCs) were cultured, and sh-Smurf2 and sh-NC vectors were transfected into GMCs induced by high glucose. Cell proliferation in blank control group (NG), high glucose treatment group (HG), high glucose + sh-NC group (HG + sh-NC), and high glucose + sh-Smurf2 group (HG + sh-Smurf2) were detected by CCK-8 and EdU. The expressions of Fibronectin, Collagen I and α-SMA proteins were detected by Western blot. CO-IP assay was employed to detect the targeting interaction between Smurf2 and EYA2, and then sh-EYA2 and sh-NC vectors were used to transfect GMCs for backtracking experiments. Results Compared with NG group, cell proliferation and protein expression of Fibronectin, Collagen I and α-SMA in HG group were significantly increased. Compared with HG group, interference with Smurf2 inhibited cell proliferation and the expression of Fibronectin, Collagen I and α-SMA proteins. CO-IP results showed a targeted interaction between Smurf2 and EYA2. Compared with HG + sh-Smurf2 group, interference with EYA2 promoted cell proliferation and the expression of Fibronectin, Collagen I and α-SMA proteins. Conclusion Interference with Smurf2 can inhibit hyperproliferation and fibrosis of GMCs induced by high glucose by upregulating EYA2.

关键词

肾小球系膜细胞 / Smurf2 / EYA2 / 增殖 / 纤维化

Key words

glomerular mesangial cells / Smurf2 / EYA2 / proliferation / fibrosis

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基金资助

漳州市自然科学基金（ZZ2023J09）; 福建省自然科学基金（2024J011586）