钙敏感受体在胰岛素抵抗状态下肾动脉平滑肌细胞增殖和迁移中的作用  

Role of calcium sensing receptors in proliferation and migration of renal artery smooth muscle cells under insulin resistance

作　　者：李光伟[1] 李波[1] 金莉[1] 邱丽萍[2] 张亚珍[2] 吴淑琴[3] LI Guang-wei;LI Bo;JIN Li;QIU Li-ping;ZHANG Ya-zhen;WU Shu-qin(Dept of Pathophysiology,Qiqihar Medical University,Qiqihar Heilongjiang161006,China;Dept of Functional Experiment,Qiqihar Medical University,Qiqihar Heilongjiang161006,China;Dept of Pathology,Qiqihar Medical University,Qiqihar Heilongjiang161006,China)

机构地区：[1]齐齐哈尔医学院病理生理学教研室,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院机能实验教学中心,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院病理学教研室,黑龙江齐齐哈尔161006

出　　处：《中国药理学通报》2021年第1期62-68,共7页Chinese Pharmacological Bulletin

基　　金：黑龙江省教育厅省属本科高校基本科研业务费项目(No 2017-QYKYYWF-0745)。

摘　　要：目的探讨钙敏感受体(calcium sensing receptor,CaSR)在胰岛素抵抗状态下肾动脉平滑肌细胞(renal artery smooth muscle cell,RAMSC)增殖和迁移中的作用及机制。方法将对数生长期的RASMC随机分为:对照组、单纯模型组、模型+GdCl 3(CaSR激动剂)组、模型+NPS2143(CaSR抑制剂)组。应用α平滑肌肌动蛋白免疫组化染色鉴定RAMSC;Western blot检测CaSR、PCNA及MMP-2蛋白的表达;采用流式细胞术检测细胞增殖周期变化;应用Transwell小室法检测细胞迁移能力;应用ELISA法检测MMP-2蛋白的含量;应用激光共聚焦扫面显微镜技术检测细胞中钙离子浓度的变化。结果与对照组比较,模型组RAMSC CaSR蛋白表达增多,PCNA及细胞增殖指数增加,细胞穿膜数量及MMP-2表达增多,同时细胞内钙离子浓度升高。GdCl 3及NPS2143分别上调和下调胰岛素抵抗引起的上述作用。结论胰岛素抵抗活化的CaSR能够促进大鼠RASMC的增殖与迁移过程,细胞内钙离子在其中发挥重要作用。Aim To investigate the role and mechanism of calcium-sensing receptor(CaSR)in the proliferation and migration of renal artery smooth muscle cells(RASMCs)under insulin resistance.Methods RAMSCs in the logarithmic growth stage were randomly divided into control,pure model,model+GdCl 3(CaSR agonist),model+NPS2143(CaSR inhibitor)group.RASMC was identified byαsmooth muscle actin immunohisitochemical staining.The expressions of CaSR,PCNA and MMP-2 were detected by Western blot.The cell proliferation cycle was detected by flow cytometry.The ability of cell migration was measured by transwell assay.The content of MMP-2 protein was detected by ELISA.The change of intracellular calcium concentration was detected by laser confocal scanning microscopy.Results Compared with control group,in model group the protein expression of CaSR increased,the cell proliferation index and PCNA were enhanced,the protein levels of MMP-2 and the number of migrated cells increased,and intracellular calcium concentration was elevated.GdCl 3 and NPS2143 up-regulated and down-regulated the above effects caused by insulin resistance,respectively.Conclusions The CaSR activated by insulin resistance can promote proliferation and migration of rat RAMSCs,in which intracellular calcium ion play an important role.

关 键 词：钙敏感受体 肾动脉平滑肌细胞 高糖 高胰岛素 增殖 迁移 

分 类 号：R-332[医药卫生—人体解剖和组织胚胎学] R322.121[医药卫生—基础医学]