IgA肾病细胞模型中的炎症表达及丙戊酸钠的抗炎作用  

Inflammatory expression in IgA nephropathy cell model and anti-inflammatory effect of sodium valproate

作　　者：戴芹 王伟铭[2] DAI Qin;WANG Weiming(Department of Nephrology,Xuhui hospital,Fudan University,Shanghai 200031,China;Department of Nephrology,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]复旦大学附属徐汇医院肾内科,上海200031 [2]上海交通大学医学院附属瑞金医院肾内科,上海200025

出　　处：《上海交通大学学报（医学版）》2022年第6期751-757,共7页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市高级中西医结合人才培养项目[ZY(2018-2020)-RCPY-2020];上海市徐汇区卫生健康系统人才培养项目(xhxtrc2019-2021)。

摘　　要：目的·研究来源于IgA肾病患者的聚集性IgA1(P-aIgA1)对人肾系膜细胞(human renal mesangial cells,HMCs)分泌炎症因子及细胞增殖的影响,并观察组蛋白去乙酰化酶(histone deacetylation,HDAC)抑制剂丙戊酸钠(sodium valproate,VPA)体外抗炎和抗细胞增殖作用。方法·应用亲和层析法制备N-IgA1(来自健康对照者的IgA1)和P-IgA1(来自IgA肾病患者的IgA1),用热聚合法制备P-aIgA1和N-aIgA1(来自健康对照者的聚集性IgA1)。取HMCs,加入不同类型的IgA1及同体积的PBS(对照组),或者VPA干预。应用人炎症因子蛋白芯片检测各组HMCs培养上清液中炎症因子表达。用MTT法检测HMCs增殖情况。采用免疫印迹法或ELISA法检测相关蛋白的表达。结果·炎症因子蛋白芯片结果显示:与对照组比较,P-aIgA1组HMCs培养上清液中白介素-6可溶性受体(interleukin-6 soluble receptor,IL-6sR)、受激活调节的正常T细胞表达和分泌因子(regulated upon activation normal T cell expressed and secreted factor,RANTES)、金属蛋白酶组织抑制因子1(tissue inhibitor of metalloproteinase 1,TIMP1)、金属蛋白酶组织抑制因子2(tissue inhibitor of metalloproteinase 2,TIMP2)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和肿瘤坏死因子Ⅱ型受体(tumor necrosis factor typeⅡreceptor,TNFRⅡ)的表达水平均显著上调,分别为对照组的34倍、124倍、269倍、100倍、1.6倍和52倍;与对照组比较,N-aIgA1,P-IgA1,P-aIgA1都能促进TIMP2的表达,差异均有统计学意义(P=0.000,P=0.000,P=0.001)。用MTT法观察HMCs的增殖程度,结果显示:①与对照组相比,N-IgA1组对HMCs的增殖无显著的影响,P-IgA1及PaIgA1都能显著促进HMCs的增殖(P=0.045和P=0.003);与N-IgA1相比,P-aIgA1组HMCs出现了显著的增殖,差异有统计学意义(P=0.036)。②不同浓度的P-aIgA1对HMCs增殖影响的结果显示:25μg/mL的P-aIgA1就可以显著促进HMCs的增殖(P=0.038),呈剂量依赖性。③400μg/mL的VPA能够显著抑制HMCs的增殖(Objective·To study the effects of aggregated IgA1(P-aIgA1)from patients with IgA nephropathy on the secretion of inflammatory factors and cell proliferation of human renal mesangial cells(HMCs),and observe the anti-inflammatory and antiproliferation effects of histone deacetylation(HDAC)inhibitor sodium valproate(VPA)in vitro.Methods·N-IgA1(IgA1 from health control)and P-IgA1(IgA1 from patients with IgA nephropathy)were prepared by affinity chromatography,and P-aIgA1 and N-aIgA1 were prepared by thermal polymerization.The expression of inflammatory factors in HMCs culture supernatant was detected by human inflammatory factor protein chip.The proliferation of HMCs was detected by MTT assay.The expression of related proteins was detected by Western blotting or ELISA.Results·The results of inflammatory factor protein chip showed that the concentration of interleukin-6 soluble receptor(IL-6sR),regulated upon activation normal T cell expressed and secreted factor(RANTES),tissue inhibitor of metalloproteinase 1(TIMP1),tissue inhibitor of metalloproteinase 2(TIMP2),tumor necrosis factor-α(TNF-α)and tumor necrosis factor type Ⅱ receptor(TNFRⅡ)in the culture supernatant of HMCs in the P-aIgA1 group were significantly up-regulated compared with the control group,which were 34,124,269,100,1.6 and 52 times higher than those in the control group,respectively;N-aIgA1,P-IgA1 and P-aIgA1 could promote the expression of TIMP2,and there were significant differences(P=0.000,P=0.000,P=0.001).The proliferation of HMCs was observed by MTT method.The results showed that:①Compared with the control group,N-IgA1 group had no significant effect on the proliferation of HMCs,and P-IgA1 and P-aIgA1 could significantly promote the proliferation of HMCs(P=0.045 and P=0.003);compared with the N-IgA1,HMCs in the P-aIgA1 group had significant proliferation,and the differences was statistically significant(P=0.036).②The results of the effects of different concentrations of P-aIgA1 on the proliferation of HMCs showed that:25μg/mL P-aIg

关 键 词：IGA肾病 人系膜细胞 组蛋白去乙酰化酶 丙戊酸钠 炎症因子 

分 类 号：R593.9[医药卫生—内科学]