脂联素对高糖条件下RF/6A细胞的保护作用  

Protective effect of adiponectin on RF/6A cells under high glucose conditions

作　　者：李蓉[1] 杜军辉[2] 姚国敏[1] 王小娣[1] 姚杨[3] LI Rong;DU Junhui;YAO Guomin;WANG Xiaodi;YAO Yang(Department of Ophthalmology,First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,China;Department of Ophthalmology,Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University;Department of Central Laboratory,First Affiliated Hospital of Xi'an Medical University)

机构地区：[1]西安医学院第一附属医院眼科,西安710077 [2]西安交通大学附属西安市第九医院眼科 [3]西安医学院第一附属医院中心实验室

出　　处：《山西医科大学学报》2019年第2期169-174,共6页Journal of Shanxi Medical University

基　　金：陕西省卫生健康科研基金项目(2018D074);西安市科技局医学研究项目(201805097YX5SF31(4));国家自然科学基金资助项目(81500726)

摘　　要：目的观察脂联素(adiponectin,APN)对高糖培养条件下恒河猴脉络膜视网膜血管内皮细胞(monkey choroid-retinalendothelial cell line,RF/6A)增殖及凋亡的影响。方法将体外培养的生长良好的RF/6A细胞随机分为空白对照组(PBS)、甘露醇对照组(25 mmol/L甘露醇)、高糖组(25 mmol/L D-葡萄糖)、高糖+APN组(25 mmol/L D-葡萄糖+10μg/mlAPN),各组加入不同的药物处理48 h。分别采用CCK-8法检测细胞增殖,Western blot检测细胞凋亡关键蛋白Bax、Bcl-2的表达,流式细胞术检测细胞凋亡率。结果高糖组(71. 42%±2. 29%)和高糖+APN组(90. 87%±1. 68%)细胞增殖率低于空白对照组(100%±0%)和甘露醇对照组(99. 09%±0. 46%)(P <0. 001);与高糖组相比,高糖+APN组细胞增殖率明显升高(P <0. 001)。高糖组和高糖+APN组的RF/6A细胞Bax的相对表达量高于空白对照组和甘露醇对照组(P <0. 001)。高糖组和高糖+APN组细胞Bcl-2的相对表达量低于空白对照组和甘露醇对照组(P <0. 001)。与高糖组相比,高糖+APN组细胞Bax的表达明显降低,Bcl-2的表达明显升高(均P <0. 001)。高糖组及高糖+APN组细胞凋亡率均比空白对照组和甘露醇对照组增多(P <0. 001)。与高糖组相比,高糖+APN组细胞凋亡率明显减少(P <0. 001)。结论 APN处理可抑制高糖培养条件下的RF/6A细胞凋亡,促进细胞增殖,提示APN对视网膜血管内皮细胞具有一定的保护效应。Objective To observe the effect of adiponectin(APN)on proliferation and apoptosis of monkey choroid-retinal endothelial cell line(RF/6A)under the condition of high glucose.Methods Well-cultured RF/6A cells in vitro were randomly divided into four groups:blank control group(PBS),mannitol control group(25 mmol/L mannitol),high glucose group(25 mmol/L D-glucose),high glucose+APN group(25 mmol/L D-glucose+10μg/ml APN).Each group was treated with different drugs for 48 h.Cell proliferation was detected by CCK-8 assay,the expression of key apoptotic proteins Bax and Bcl-2 was detected by Western blot,and the cell apoptosis rate was detected by flow cytometry.Results The proliferation rate in high glucose group(71.42%±2.29%)and high glucose+APN group(90.87%±1.68%)was lower than that in blank control group(100%±0%)and mannitol control group(99.09%±0.46%)(all P<0.001).Compared with high glucose group,the cell proliferation rate in high glucose+APN group was significantly increased(P<0.001).The expression of Bax in both high-glucose group and high-glucose+APN group was higher than that in blank control group and mannitol control group(P<0.001).The expression of Bcl-2 in both high-glucose group and high-glucose+APN group was lower than that in blank control group and mannitol control group(P<0.001).Compared with high glucose group,the expression of Bax was significantly decreased in high glucose+APN group,while the expression of Bcl-2 was significantly increased(all P<0.001).The apoptosis rate in high glucose group and high glucose+APN group was higher than that in blank control group and mannitol control group(P<0.001).The apoptosis rate was significantly reduced in high glucose+APN group compared with high glucose group(P<0.001).Conclusion APN can inhibit apoptosis of RF/6A cells and promote cell proliferation under high glucose conditions,suggesting that APN has a certain protective effect on retinal vascular endothelial cells.

关 键 词：脂联素 高糖培养 视网膜血管内皮细胞 凋亡 细胞增殖 

分 类 号：R587[医药卫生—内分泌]