钙敏感受体在高糖诱导的人视网膜微血管内皮细胞内皮间质转化中的作用

Role of the calcium-sensing receptor in the high glucose-induced endothelial-mesenchymal transition of human retinal capillary endothelial cells

目的观察钙敏感受体(CaSR)在高糖(HG)诱导的人视网膜血管内皮细胞(HRCECs)内皮间质转化(EndMT)中的作用和机制。方法将HRCECs分为3组:对照组(NG组,培养基中加入5.5 mmol·L^(-1)D-葡萄糖)、高糖组(HG组,培养基中加入25.0 mmol·L-1 D-葡萄糖)及HG+Calhex231组(培养基中加入25.0 mmol·L^(-1)D-葡萄糖和3μmol·L^(-1)Calhex231)。CCK-8法检测各组HRCECs的细胞活力,Transwell实验检测各组HRCECs的迁移能力,利用细胞管腔形成实验检测各组HRCECs的管腔形成能力,Western blot检测CaSR、VE-cadherin、CD31、α-SMA及Nuclearβ-catenin蛋白的表达。结果HG组CaSR的表达较NG组增加。与NG组相比,HG组HRCECs的细胞活力、迁移能力和成管能力均增强,细胞中CaSR、α-SMA和Nuclearβ-catenin蛋白表达均明显增加,VE-cadherin和CD31蛋白表达均明显降低(均为P<0.05)。与HG组相比,HG+Calhex231组HRCECs的上述指标均明显逆转(均为P<0.05)。结论CaSR表达参与HG诱导的EndMT进展,其机制可能与促进β-catenin核易位有关。

Objective To observe the role and mechanism of calcium-sensing receptor(CaSR)in high glucose(HG)-induced endothelial-mesenchymal transition(EndMT)of human retinal capillary endothelial cells(HRCECs).Methods The HRCECs were divided into the control group(NG group,5.5 mmol·L^(-1)glucose was given),high glucose group(HG group,25.0 mmol·L^(-1)glucose was given)and HG+Calhex231 group(25.0 mmol·L^(-1)glucose and 3μmol·L-1 Calhex231 were given).Cell Counting Kit-8 was used to measure the viability of HRCECs in each group.The migration ability of HRCECs was detected by the Transwell assay,and the angiogenesis of HRCECs was measured by tube formation assay.The expression levels of CaSR,vascular endothelial(VE)-cadherin,cluster of differentiation 31(CD31),alpha-smooth muscle actin(α-SMA)and Nuclearβ-catenin were detected by Western blot.Results The expression of CaSR in the HG group was higher than that in the NG group.Compared with the NG group,the viability,migration,and angiogenesis ability of HRCECs and the expressions of CaSR,α-SMA and Nuclearβ-catenin significantly increased,while the expressions of VE-cadherin and CD31 significantly decreased in the HG group(all P<0.05).Compared with the HG group,the above indexes in the HG+Calhex231 group were significantly reversed(all P<0.05).Conclusion The CaSR expression is involved in HG-induced EndMT of HRCECs,which may be related to the promotion ofβ-catenin nuclear translocation.