抑瘤素M在高血糖致人主动脉平滑肌细胞增殖中的作用  

Effects on oncostatin M in hyperglycemia-induced aortic smooth muscle cells proliferation in human

作　　者：李连喜[1] 陶征[2] 梁文昌[3] 杨志红[3] 周闻白[3] 叶威巍[3] 包玉倩[1] 贾伟平 胡仁明[3] 

机构地区：[1]上海市糖尿病研究所,上海市糖尿病临床医学中心,上海市糖尿病重点实验室,上海交通大学附属第六人民医院内分泌代谢科,200233 [2]江苏大学附属人民医院信息科 [3]复旦大学附属华山医院内分泌科,复旦大学内分泌糖尿病研究所

出　　处：《中华糖尿病杂志》2009年第3期-,共5页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：上海市卫生局科研项目，复旦大学博士创新基金

摘　　要：目的 研究高血糖对人巨噬细胞中抑瘤素M表达的影响以及抑瘤素M对人主动脉平滑肌细胞增殖的影响.方法 采用佛波酯孵育人单核/巨噬细胞系THP-1细胞,诱导其分化为巨噬细胞.应用人细胞因子抗体芯片筛选出受高血糖(15 mmol/L)调控的人巨噬细胞源性细胞因子,并经Western blot和酶联免疫吸附试验(ELISA)技术验证.运用逆转录-聚合酶链反应(RT-PCR)明确人主动脉内皮细胞、平滑肌细胞及巨噬细胞中是否存在抑瘤素M受体GP130-OSMRβ二聚体及GP130-LIFR二聚体表达.运用细胞增殖实验(XTT法)研究抑瘤素M对人主动脉平滑肌细胞增殖的影响.采用单因素方差分析进行统计学分析.结果 佛波酯孵育48 h可诱导THP-1细胞分化为巨噬细胞.细胞因子芯片结果显示高血糖可上调抑瘤素M在巨噬细胞中的表达,并为Western blot及ELISA所证实.RT-PCR显示人主动脉内皮细胞、平滑肌细胞及巨噬细胞均可表达GP130-OSMRβ二聚体及GP130-LIFR二聚体.XTT法证实抑瘤素M可促进血管平滑肌细胞增殖,与0μg/L抑瘤素M(吸光度为0.468±0.032)相比,5μg/L(吸光度为0.503±0.026)、10μg/L(吸光度为0.520±0.027)、20μg/L抑瘤素M(吸光度为0.513±0.026)人主动脉平滑肌细胞增殖均明显增加(F=10.40,P<0.05).结论 高血糖可促进巨噬细胞中抑瘤素M的表达和分泌.抑瘤素M通过与人主动脉平滑肌细胞中的受体结合而促进血管平滑肌细胞的增殖,进而部分参与糖尿病大血管病变的发生发展.Objective To study the effects of high blood glucose on expression of oncostatin M (OSM) in macrophages and the association of OSM with proliferation of human aortic smooth muscle cells. Methods Monocytic cell line THP-1 was incubated with PMA to induce macrophages. The human macrophage cell line-derived cytokines influenced by hyperglycemia (15 mmol/L) were screened by human cytokine antibody array, which was further confirmed by Western blot and ELISA. RT-PCR was used to determine the expression of OSM receptors GP130-OSMRβ and GP130-LIFR in human aortic endothelial cells, smooth muscle cells and macrophages. The effects of OSM on proliferation of human aortic smooth muscle cell was assessed using the XTT assay. One-way analysis of variances was used for data analysis. Results Incubation with PMA induced mature macrophages. Human cytokine antibody array showed that high blood glucose up-regulated the expression of OSM in macrophages, which was further confirmed by Western blot and ELISA. The expression of GPI30-OSMRβ and GP130-LIFR was found in human aortic endothelial cells, smooth muscle cells and macrophages. XTT assay showed that OSM promoted proliferation of human aortic smooth muscle cells. Compared with 0 μg/L OSM (OD = 0.468 ±0.032), the proliferation of human aortic smooth muscle cells was significantly increased in 5, 10, 20 μg/L OSM (OD = 0.503±0.026, 0.520±0.027, 0.513±0.026, respectively) (F=10.40,P<0.05) Conclusions High blood glucose could increase the expression and secretion of OSM in macrophages. OSM may promote the proliferation of human aortic smooth muscle cells by binding with its receptors, which partly involves in the development of diabetic macroangiopathy.

关 键 词：糖尿病 巨噬细胞 肌 平滑 血管 抑瘤素M 

分 类 号：R54[医药卫生—心血管疾病] R587[医药卫生—内科学]