钙敏感受体在高糖诱导的人眼小梁网细胞损伤中的作用  

Role of calcium-sensing receptor in high glucose-induced injury of hu⁃man trabecular meshwork cells

作　　者：车冰冰 孙梦婷 王宁宁 岑杰 李国鹏 赵冰冰 魏璨[1] CHE Bing-bing;SUN Meng-ting;WANG Ning-ning;CEN Jie;LI Guo-peng;ZHAO Bing-bing;WEI Can(Department of Pathophysiology,Harbin Medical University,Harbin 150086,China;Affiliated Second Hospital,Harbin Medical University,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学病理生理学教研室,黑龙江哈尔滨150086 [2]哈尔滨医科大学附属第二医院,黑龙江哈尔滨150086

出　　处：《中国病理生理杂志》2022年第5期839-844,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82170268;No.81800260);黑龙江省自然科学基金(No.YQ2021H004);黑龙江省博士后启动金(No.LBH-Q21024)。

摘　　要：目的:观察钙敏感受体(CaSR)在高糖(HG)诱导的人眼小梁网细胞(HTMC)损伤中的作用和机制。方法:将HTMC随机分为:对照(control)组(10%胎牛血清-DMEM)、HG组(10%胎牛血清-DMEM+50 mmol/L葡萄糖)、HG+CaSR激动剂NPS R568组(10%胎牛血清-DMEM+50 mmol/L葡萄糖+10μmol/L NPS R568)和HG+CaSR抑制剂Calhex231组(10%胎牛血清-DMEM+50 mmol/L葡萄糖+3μmol/L Calhex231),培养72 h。CCK-8法测量不同浓度的葡萄糖刺激下HTMC活力;试剂盒测定超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;Western blot检测CaSR、SOD2、凋亡相关蛋白(cleaved caspase-3和Bcl-2)及自噬相关蛋白(ATG7、P62、LC3-I和LC3-Ⅱ)表达;免疫荧光染色观察CaSR细胞定位和表达。结果:葡萄糖浓度为50 mmol/L时,HTMC相对活力由1.00±0.03下降至0.60±0.07(P<0.05);与control组相比,HG使HTMC中CaSR、cleaved caspase-3和P62表达以及MDA含量增加(P<0.05),而SOD活性,SOD2、Bcl-2和ATG7表达,以及LC3-Ⅱ/LC3-I比值下降(P<0.05);NPS R568进一步促进HG诱导的细胞损伤,而Calhex231可减轻上述变化(P<0.05)。结论:CaSR表达参与HG诱导的HTMC损伤,其机制与促进细胞凋亡和氧化应激以及抑制自噬有关。AIM:To observe the role and mechanism of calcium-sensing receptor(CaSR)in high glucose(HG)-induced human trabecular meshwork cell(HTMC)injury.METHODS:The HTMC were randomly divided into control group[10% fetal bovine serum(FBS)-DMEM culture],HG group(10% FBS-DMEM+50 mmol/L glucose),HG+CaSR agonist NPS R568 group(10% FBS-DMEM+50 mmol/L glucose+10 μmol/L NPS R568)and HG+CaSR inhibitor Calhex231 group(10% FBS-DMEM+50 mmol/L glucose+3 μmol/L Calhex231). The treatments lasted for 72 h. CCK-8assay was used to measure the viability of HTMC stimulated by different concentrations of glucose. The activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)were measured by kits. The expression levels of CaSR,SOD2,apoptosis-related proteins(cleaved caspase-3 and Bcl-2)and autophagy-related proteins(ATG7,P62,LC3-I and LC3-Ⅱ)were measured by Western blot. The localization and expression of CaSR were detected by immunofluorescence.RESULTS:The relative HTMC viability was significantly decreased by glucose at a concentration of 50 mmol/L from 1. 00±0. 03 to 0. 60±0. 07(P<0. 05). Compared with control group,HG increased the expression of CaSR,cleaved caspase-3 and P62,and the content of MDA(P<0. 05),but decreased the expression of Bcl-2,SOD2 and ATG7,the ratio of LC3-Ⅱ/LC3-I,and the activity of SOD(P<0. 05). Treatment with NPS R568 further enhanced,but Calhex231 alleviated HG-induced cell injury(P<0. 05).CONCLUSION:The CaSR expression is involved in HG-induced HTMC injury partially by promoting apoptosis and oxidative stress and inhibiting autophagy.

关 键 词：钙敏感受体 人小梁网细胞 高糖 细胞凋亡 氧化应激 自噬 

分 类 号：R363[医药卫生—病理学]