补体因子H相关蛋白1促进巨噬细胞分泌肿瘤坏死因子-α调控足细胞增殖和迁移实验研究  

Complement factor H-related protein 1 promotes the secretion of TNF-α by macrophages and regulates podocyte proliferation and migration Experimental study

作　　者：莫颖[1] 王凤梅 帕提古丽·阿斯讨拜[1] 欧云塔娜 MO Ying;WANG Fengmei;ASTAOBAI Patiguli;OUYUN Tana(Department of Nephrology,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 820011,China)

机构地区：[1]新疆医科大学第五附属医院肾病科,新疆乌鲁木齐820011 [2]东南大学附属中大医院肾病科,江苏南京210009 [3]新疆医科大学第五临床医学院,新疆乌鲁木齐820011

出　　处：《陕西医学杂志》2024年第4期444-448,共5页Shaanxi Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(2021D01C434)。

摘　　要：目的:探讨补体因子H相关蛋白1(CFHR1)通过巨噬细胞分泌肿瘤坏死因子-α(TNF-α)调控足细胞增殖和迁移的作用。方法:体外培养人单核巨噬细胞和人肾足细胞。巨噬细胞分为对照组和CFHR1干预组,分别进行牛血清白蛋白或CFHR1重组蛋白干预24 h,ELISA法测定上清液TNF-α水平。足细胞分为空白组、TNF-α干预组、对照上清液干预组、CFHR1上清液干预组、CFHR1上清液+TNF-α中和抗体干预组。CCK8法检测各组细胞增殖。Transwell法检测各组细胞迁移。Wb法检测各组细胞中相关蛋白变化。结果:巨噬细胞的CFHR1干预组上清液中TNF-α含量显著增加(P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的细胞增殖比率和迁移数量均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的细胞增殖比率和迁移数量均显著提高(均P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的足细胞裂孔膜蛋白(Nephrin)、足突蛋白(Podocin)、纤维状肌动蛋白(F-Actin)、整合素α3β1蛋白(α3β1)表达均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的Nephrin、Podocin、F-actin、α3β1蛋白表达均显著增多(均P<0.05)。结论:CFHR1促进巨噬细胞分泌的TNF-α可显著抑制足细胞增殖水平和迁移能力,这可能是高浓度CFHR1促进肾病综合征发展的途径。Objective:To investigate the role of complement factor H-related protein 1(CFHR1)in regulating podocyte proliferation and migration through TNF-α secreted by macrophages.Methods:Cultivate human monocyte macrophages and human renal podocytes in vitro.Macrophages were divided into control group and CFHR1 intervention group and treated with bovine serum albumin or CFHR1 recombinant protein for 24 hours,respectively.The supernatant TNF-α was measured using ELISA method.Podocytes are divided into blank group,TNF-α intervention group,control supernatant intervention group,CFHR1 supernatant intervention group,CFHR1 supernatant+TNF-α neutralizing antibody intervention group.The CCK8 method was used to detect cell proliferation.Transwell method was used to detect cell migration.Western blot was used to detect changes in related proteins of cells.Results:The TNF-α content in the supernatant of the CFHR1 intervention group of macrophages was significantly increased(P<0.05).Compared with the blank group,the cell proliferation rate and migration number in the TNF-α intervention group and CFHR1 supernatant intervention group were significantly reduced(all P<0.05).Compared with the CFHR1 supernatant intervention group,the cell proliferation rate and migration number of the CFHR1 supernatant+TNF-α neutralizing antibody intervention group were significantly increased(all P<0.05).Compared with the blank group,the protein expression of Nephrin,Podocin,F-actin,andα3β1 in the TNF-α intervention group and CFHR1 supernatant intervention group was significantly reduced(all P<0.05).Compared with the CFHR1 supernatant intervention group,the protein expression of Nephrin,Podocin,F-actin,and α3β1 in the CFHR1 supernatant+TNF-α neutralizing antibody intervention group was significantly increased(all P<0.05).Conclusion:CFHR1 promotes macrophages secretion of TNF-α,and significantly inhibits the proliferation level and migration ability of podocytes,which may be a pathway for high concentration CFHR1 to promote the development of

关 键 词：补体因子H相关蛋白1 肿瘤坏死因子-Α 足细胞 巨噬细胞 增殖 迁移 

分 类 号：R73-3[医药卫生—肿瘤]