补阳还五汤对高糖环境下miR-21介导的人视网膜微血管内皮细胞自噬的影响  

Effect of Buyang Huanwu decoction on miR-21-mediated autophagy in human retinal capillary endothelial cells under high glucose

作　　者：石颖 陈子扬 陈凯铭 胡艳红[3] 胡俊[3] 陈胜[3] SHI Ying;CHEN Ziyang;CHEN Kaiming;HU Yanhong;HU Jun;CHEN Sheng(Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Comprehensive Outpatient Department of Fujian Academy of Chinese Medicine,Fuzhou 350003,Fujian Province,China;The Second Affiliated Hospital of Fujian Traditional Chinese Medical University,Fuzhou 350003,Fujian Province,China)

机构地区：[1]福建中医药大学,福建省福州市350122 [2]福建省中医药科学院综合门诊部,福建省福州市350003 [3]福建中医药大学附属第二人民医院眼科,福建省福州市350003

出　　处：《眼科新进展》2022年第11期858-862,共5页Recent Advances in Ophthalmology

基　　金：福建省卫健委科技计划项目(编号:2020CXB034);福建省自然科学基金计划项目(编号:2022J01834)。

摘　　要：目的观察补阳还五汤对高糖环境下miR-21介导的人视网膜微血管内皮细胞(HRCECs)自噬的影响,以期为DR自噬调控机制及药物治疗提供依据。方法HRCECs随机分5组,即正常对照组、高糖组、补阳还五汤组、miR-21 mimic组、miR-21 mimic+补阳还五汤组。除正常对照组外,其余各组细胞于25.0 g·L^(-1)葡萄糖环境中培养,建立高糖损伤模型,补阳还五汤组加用5 g·L^(-1)补阳还五汤水提液干预;miR-21 mimic组孵育转染miR-21 mimic的HRCECs;miR-21 mimic+补阳还五汤组用5 g·L^(-1)补阳还五汤水提液干预转染miR-21 mimic的HRCECs。荧光显微镜下观察转染效率,实时荧光定量PCR检测各组细胞中miR-21的表达,Western blot检测各组细胞中LC3蛋白表达。结果经转染miR-21 mimic后,大部分HRCECs呈绿色荧光,荧光强度为32.708±1.001。实时荧光定量PCR检测结果显示,miR-21 mimic组HRCECs的miR-21相对表达量为106.677±5.787,明显高于高糖组(5.892±0.341)和正常对照组(0.993±0.015),差异均有统计学意义(均为P<0.05)。高糖组和miR-21 mimic组HRCECs中LC3蛋白相对表达量均较正常对照组升高,差异均有统计学意义(均为P<0.05),且miR-21 mimic组HRCECs中LC3蛋白相对表达量高于高糖组(P=0.001)。补阳还五汤组HRCECs中LC3蛋白相对表达量低于高糖组和miR-21 mimic组(均为P<0.05),与正常对照组比较差异无统计学意义(P=0.197)。miR-21 mimic+补阳还五汤组HRCECs中LC3蛋白相对表达量低于miR-21 mimic组,但高于补阳还五汤组,差异均有统计学意义(均为P<0.05),与高糖组比较差异无统计学意义(P=0.930)。结论高糖环境下,miR-21介导HRCECs的自噬,补阳还五汤可以下调HRCECs的miR-21表达,进而减少细胞自噬。Objective To investigate the effect of Buyang Huanwu decoction(BHD)on miR-21-mediated autophagy in human retinal capillary endothelial cells(HRCECs)under high glucose(HG)to provide a basis for research on the DR autophagy regulation mechanism and drug therapy.Methods HRCECs were randomly divided into five groups:normal control(NC)group,HG group,BHD group,miR-21 mimic group,and miR-21 mimic+BHD group.Cells were cultured in 25 g·L^(-1) glucose and HG damage models were established except the NC group.In addition,cells in the BHD group were intervened with 5 g·L^(-1) BHD water extract;cells in the miR-21 mimic group were transfected with miR-21 mimic;cells in the miR-21 mimic+BHD group were transfected with miR-21 mimic and then were intervened with 5 g·L^(-1) BHD water extract.Transfection efficiency was assessed under the fluorescence microscope.The miR-21 expression in cells was detected by quantitative real-time polymerase chain reaction(qRT-PCR).The LC3 protein expression was measured by Western blot.Results After transfection with miR-21 mimic,most HRCECs showed green fluorescence,with an average fluorescence intensity of 32.708±1.001.The qRT-PCR results showed that the relative expression of miR-21 in the miR-21 mimic group was 106.677±5.787,significantly higher than that in the HG group(5.892±0.341)and NC group(0.993±0.015)(both P<0.05).LC3 protein expression rose in both the HG and miR-21 mimic groups compared with the NC group(both P<0.05),and LC3 protein expression in the miR-21 mimic group was higher than that in the HG group(P=0.001).LC3 protein expression in the BHD group was lower than that in the HG and miR-21 mimic groups(both P<0.05),but showed no significant difference compared with the NC group(P=0.197).LC3 protein expression in the miR-21 mimic+BHD group was lower than that in the miR-21 mimic group,but higher than that in the BHD group(both P<0.05).There was no significant difference in LC3 protein expression between the miR-21 mimic+BHD group and the HG group(P=0.930).Conclusion miR-21

关 键 词：补阳还五汤 MIR-21 自噬 高糖 视网膜微血管内皮细胞 

分 类 号：R774.1[医药卫生—眼科]