梓醇对AGEs刺激肾系膜细胞介导巨噬细胞极化的影响

发布时间：2018-03-20 18:52

本文选题：梓醇　切入点：晚期糖基化产物　出处：《中国药理学通报》2017年10期 　论文类型：期刊论文

【摘要】：目的探讨梓醇对晚期糖基化终末产物(AGEs)刺激肾系膜细胞介导巨噬细胞极化的影响。方法将巨噬细胞(RAW264.7)置于Transwell上层小室,肾系膜细胞(MMCs)于下层孔板进行体外共培养,设置模型组、0-BSA对照组、梓醇组(0.1、1.0、10.0μmol·L~(-1)),另设氨基胍组(1.0μmol·L~(-1))作为阳性对照。加入各药物预孵下层系膜细胞1 h后,用AGEs(100 mg·L~(-1))刺激,继续孵育23 h,采用ELISA法检测系膜细胞上清液中单核细胞趋化蛋白-1(MCP-1)的分泌水平;Western blot法检测巨噬细胞M1型标记蛋白i NOS、CD16/32、TNF-α、COX-2和M2型标记蛋白CD206、Arg-1的表达水平;流式细胞仪检测巨噬细胞i NOS与CD206蛋白表达百分率。结果 AGEs可提高系膜细胞MCP-1的分泌水平(P0.01),上调巨噬细胞i NOS、TNF-α、CD16/32、COX-2蛋白表达(P0.05,P0.01),下调CD206和Arg-1蛋白表达;而梓醇(0.1、1.0、10.0μmol·L~(-1))预孵能降低系膜细胞MCP-1的分泌水平(P0.01),下调i NOS、TNF-α、CD16/32、COX-2蛋白表达(P0.05,P0.01),上调CD206和Arg-1蛋白表达(P0.05),并呈浓度依赖性。结论梓醇可通过调控AGEs刺激系膜细胞分泌MCP-1的水平,抑制巨噬细胞的M1型极化,并促进其向M2型极化,减轻炎症反应,缓解糖尿病肾损伤。
[Abstract]:Objective to investigate the effect of catalpol on the late glycation end product (ages) -stimulated renal Mesangial cell mediated macrophage polarization. Methods macrophages (RAW264.7) were placed in the upper chamber of Transwell, and Mesangial cells (MMC) were co-cultured in the lower pore plate in vitro. The model group was divided into 0 BSA control group, catalpol group (0.1 渭 mol 路L ~ (-1)) and aminoguanidine group (1.0 渭 mol 路L ~ (-1)) as a positive control. The cells were preincubated with each drug for 1 h and stimulated with AGEs(100 mg 路L ~ (-1). After 23 h incubation, the secretion level of monocyte chemoattractant protein (MCP-1) in the supernatant of Mesangial cells was detected by ELISA assay. The expression of macrophage M1 labeled protein I NOSU CD16 / 32 TNF- 伪 COX-2 and M2 labeled protein CD206 (Arg-1) was detected by Western blot. Results the percentage of I NOS and CD206 protein expression in macrophages was detected by flow cytometry. Results AGEs could increase the level of MCP-1 secretion in Mesangial cells and up-regulate the expression of iNOSNF- 伪 CD16 / 32 COX-2 protein, and down-regulate the expression of CD206 and Arg-1. The preincubation of catalpol (0.1 渭 mol 路L ~ (-1)) could decrease the level of MCP-1 secretion and down-regulate the expression of P0.05 / 32COX-2 protein in Mesangial cells, and up-regulate the expression of CD206 and Arg-1 proteins in a dose-dependent manner. Conclusion Catalpol can regulate the level of MCP-1 secreted by AGEs stimulated Mesangial cells. The M 1 polarization of macrophages was inhibited, and the M 2 polarization was promoted, the inflammatory reaction was alleviated, and the kidney injury of diabetes mellitus was alleviated.
【作者单位】：南京中医药大学药学院;江苏省中药药效与安全性评价重点实验室;
【基金】：国家自然科学基金资助项目(No 81374029,81073111) 江苏省中药学优势学科开放课题(No JKLPSE201604)
【分类号】：R587.2;R692.9

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