白藜芦醇对高糖环境下视网膜细胞凋亡的抑制作用  

Inhibitory effects of resveratrol on high glucose-induced apoptosis of retinal cells

作　　者：李青春[1] 李岱[1] 邢怡桥[2] Li Qingchun;Li Dai;Xing Yiqiao(Hubei University of Science and Technology,Xianning 437100,China;Ophthalmological Center,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]湖北科技学院眼科教研室,咸宁437100 [2]武汉大学人民医院眼科中心,武汉430060

出　　处：《中华实验眼科杂志》2020年第11期929-935,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：湖北省教育厅基金资助项目(B2015074);湖北科技学院博士启动基金项目(2018-20XB013)。

摘　　要：目的观察白藜芦醇(Res)对链脲佐菌素诱发的糖尿病大鼠视网膜神经节细胞(RGCs)及高糖状态视网膜色素上皮(RPE)细胞凋亡的影响。方法取25只SD大鼠腹腔注射链脲佐菌素制作糖尿病模型,确定造模成功20只,按照随机数字表法将模型鼠分为糖尿病模型组和Res灌胃组,另选取10只大鼠作为正常对照组。Res灌胃组大鼠用Res 40 mg/(kg·d)灌胃给药,正常对照组和糖尿病模型组大鼠用等容量生理盐水灌胃,分别于灌胃后当日(0周)、4、8、12周测定大鼠体质量和经尾尖静脉采血测定血糖浓度,连续给药12周时处死大鼠并取双眼眼球,透射电子显微镜下观察各组大鼠RGCs超微结构变化;采用TUNEL法检测RGCs凋亡情况并计算凋亡指数。将ARPE-19细胞株分为正常对照组、高糖组及Res处理组,分别用含5.5 mmol/L葡萄糖、30 mmol/L葡萄糖和30 mmol/L葡萄糖+10μmol/L Res的培养基培养48 h,用流式细胞仪检测各组细胞凋亡率;采用免疫荧光法及Western blot法检测细胞中bax和bcl-2蛋白表达。结果给药4、8、12周时,Res灌胃组大鼠体质量均高于相应时间点糖尿病模型组(均P<0.01),给药后8、12周时,Res灌胃组糖浓度均低于相应时间点糖尿病模型组(均P<0.01)。透射电子显微镜下可见糖尿病模型组RGCs核染色质凝聚,线粒体肿胀及细胞质空泡化;Res灌胃组RGCs超微结构变化明显轻于糖尿病模型组。Res灌胃组RGCs层和视网膜内核层细胞凋亡指数分别为(18.36±3.37)%和(23.67±8.98)%,分别低于糖尿病模型组的(83.91±9.8)%和(64.26±10.66)%,差异均有统计学意义(均P<0.01)。正常对照组、高糖组、Res处理组ARPE-19细胞凋亡率分别为(3.11±0.26)%、(11.41±1.06)%和(5.38±0.58)%,Res处理组细胞凋亡率明显低于高糖组,差异有统计学意义(P<0.01)。高糖组细胞核中bax蛋白荧光明显增强,bcl-2蛋白荧光强度明显减弱;Res处理组细胞核bax蛋白荧光强于正常对照组,但弱于高糖�Objective To investigate the inhibitory effects of resveratrol(Res)on the apoptosis of retinal neurons of diabetic rats and ARPE-19 cells induced by high glucose.Methods Streptozotocin was intraperitoneally injected to establish a diabetes model in 25 adult male SD rats,and 20 successful models were randomized into diabetic model group and Res-administered group according to random number table method.Another 10 matched normal rats were served as normal control group.Res was intragastrically given to the rats in the Res-administered group with the dose of 40 mg/(kg·d),and an equivalent volume of normal saline solution was used in the same way in the diabetic model group and normal control group.The body weight and blood glucose level were measured on the 0th,4th,8th,12th week of administration.The rats were sacrificed on the 12th week by over-anesthesia and the eyeballs were enucleated.The ultrastructure of the retinal ganglion cells(RGCs)were examined under the transmission electron microscope.The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling(TUNEL)assay was to assess the apoptosis of retinal neurons.ARPE-19 cells were divided into normal control group,high glucose group and Res-treated group and cultured for 48 hours in medium containing 5.5 mmol/L glucose,30 mmol/L glucose,30 mmol/L glucose and 10 mol/L Res,respectively.Apoptosis rate was detected by a flow cytometry.The distribution and expression of bax and bcl-2 in the cells was detected by immunofluorescence and Western blot assay,respectively.This study protocol was approved by an Experimental Animal Ethic Committee of Hubei University of Science and Technology,and the use and care of the animals complied with the Statement of ARVO.Results Compared with the diabetic model group,the body weight was increased at 4-12 weeks and the blood glucose level was lowered at 8-12 weeks of Res administration in the Res-administered group(both at P<0.01).The chromatin condensation,mitochondrial swelling and vacuolation in cytoplasm were

关 键 词：白藜芦醇 视网膜神经节细胞 视网膜色素上皮细胞 凋亡 凋亡相关蛋白 

分 类 号：R285.5[医药卫生—中药学]