霉酚酸对小鼠肾小球系膜细胞增殖及分泌单核细胞趋化因子-1的影响

Effect of mycophenolic acid on the proliferation and MCP-1 secretion in the glomerular mesangial cells

目的 探讨霉酚酸在体外对小鼠肾小球系膜细胞增殖及其分泌单核细胞趋化因子1的影响。方法 在小鼠肾小球系膜细胞的培养基中加入不同浓度的霉酚酸(0.1、1、5 及10μmol/L),培养24 h,采用四甲基偶氮唑盐掺入法,以酶联免疫检测仪测定各组570 nm波长的吸光度值,以计算肾小球系膜细胞的存活率;另在肾小球系膜细胞的培养基中加入不同浓度的霉酚酸(0、0.5及2.5μmol/L),并同时加入肿瘤坏死因子(20 ng/ml)、白细胞介素1β(2 ng/ml)及γ干扰素(10 ng/ml)刺激4 h,收集上清液,测定单核细胞趋化因子1浓度,并行淋巴细胞迁移实验。结果 培养基中加入霉酚酸者,肾小球系膜细胞的增殖受到明显的抑制,并呈剂量依赖性;在肿瘤坏死因子、白细胞介素1β及γ干扰素的刺激下,培养的肾小球系膜细胞分泌单核细胞趋化因子1 明显增加,加入霉酚酸后,能明显抑制单核细胞趋化因子1的分泌(P<0.01),并能抑制淋巴细胞的迁移。结论 霉酚酸在体外可以抑制肾小球系膜细胞的增殖及趋化性细胞因子的分泌,这为临床上研究慢性移植肾肾病的发生和发展提供实验依据。

Objective To explore the inhibitory effect of mycophenolic acid (MPA) on the proliferation and monocyte chemoattractant protein-1 (MCP-1) secretion in mice glomerular mesangial cells (GMCs).Methods MPA with different concentrations ( 0.1 , 1, 5 and 10 μmol/L ) was added into the cultured mice GMC line in vitro for 24 h. The GMCs proliferation level was detected by the 3-(4,5-Dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) incorporation. The absorbance for counting the GMCs survival was read at 570 nm by using an automated microplate ELISA reader. The cells treated with MPA (0, 0.5 and 2.5 μmol/L ) were stimulated by TNF-α (20 ng/ml), IL-1β (2 ng/ml), IFN-γ (10 ng/ml). Four h later, the MCP-1 concentration in supernatant was determined by ELISA. At the same time, the lymphocyte migration was examined.Results MPA has a concentration -dependent antiproliferative effect on cultured GMCs. The secretion of MCP-1 of cultured GMCs stimulated by TNF-α, IL-1β, IFN-γ was significantly increased. Addition of MPA could signi -ficantly inhibit the secretion of MCP-1 ( P < 0.01 ). The lymphocyte migration was also markedly prohibited .Conclusion MPA exerts a negative effect on the proliferation of GMCs and the secretion of MCP-1 in vitro, which provides the experimental foundation for the studies of the occurrence and progression of chronic allograft nephropathy in clinical transplantation.