潜伏转化生长因子β结合蛋白2(LTBP2)对氧化型低密度脂蛋白诱导的人视网膜色素上皮细胞氧化损伤的影响  

Latent TGF-βbinding protein-2 aggravates oxidized low-density lipoproteinox-induced oxidative injury in the human retinal pigment epithelial cells

作　　者：孙连义 白淑玮 李凤至 赵梅生[2] SUN Lianyi;BAI Shuwei;LI Fengzhi;ZHAO Meisheng(Department of Ophthalmology,Xi’an People’s Hospital(Xi’an Fourth Hospital),Shaanxi Eye Hospital,Affiliated Guangren Hospital,School of Medicine,Xi’an Jiaotong University,Xi’an 710004,Shaanxi Province,China;Department of Ophthalmology,the Second Hospital of Jilin University,Changchun 130041,Jilin Province,China)

机构地区：[1]西安市人民医院(西安市第四医院),陕西省眼科医院,西安交通大学医学院附属广仁医院眼科,陕西省西安市710004 [2]吉林大学第二医院眼科,吉林省长春市130041

出　　处：《眼科新进展》2021年第6期540-544,共5页Recent Advances in Ophthalmology

摘　　要：目的探索潜伏转化生长因子β结合蛋白2(LTBP2)对氧化型低密度脂蛋白(ox-LDL)诱导的人视网膜色素上皮(ARPE)细胞氧化损伤的影响。方法0 mg·L^(-1)、50 mg·L^(-1)、100 mg·L^(-1)和200 mg·L^(-1) ox-LDL处理ARPE-19细胞,CCK-8试剂盒和细胞凋亡检测试剂盒检测细胞活力和凋亡水平,Western blot检测不同浓度ox-LDL处理后细胞LTBP2蛋白表达水平。100 mg·L^(-1) ox-LDL处理ARPE-19细胞构建氧化损伤模型。将ARPE-19细胞随机分为对照组,ox-LDL组,转染LTBP2过表达载体或阴性对照的ox-LDL+ov-LTBP2组和ox-LDL+ov-NC组,转染LTBP2 siRNA或阴性对照的ox-LDL+si-LTBP2组和ox-LDL+si-NC组,以及ox-LDL+si-LTBP2+VEGF组。相应处理各组细胞后,采用CCK-8试剂盒和细胞凋亡检测试剂盒检测细胞活力和凋亡水平,酶联免疫吸附实验检测氧化应激标志物活性氧(ROS)、超氧化物歧化酶(SOD)、血管内皮生长因子(VEGF)的表达水平,Western blot检测各组细胞LTBP2、P38和p-P38的蛋白表达水平。结果与0 mg·L^(-1) ox-LDL处理后相比,50 mg·L^(-1)、100 mg·L^(-1)和200 mg·L^(-1) ox-LDL处理后APRE-19细胞活力下降,凋亡率升高,LTBP2蛋白表达水平增加,且变化均具有剂量依赖性(均为P<0.05)。与ox-LDL+ov-NC组相比,ox-LDL+ov-LTBP2组细胞LTBP2蛋白表达水平、ROS含量及细胞凋亡率均增加,细胞活力、SOD活性均降低,p-P38/P38比值和VEGF表达量均增加(均为P<0.05)。与ox-LDL+si-NC组相比,ox-LDL+si-LTBP2组细胞LTBP2蛋白表达水平、ROS含量及细胞凋亡率均降低,细胞活力、SOD活性均增加,p-P38/P38比值和VEGF表达量均减少(均为P<0.05)。而与ox-LDL+si-LTBP2组比较,ox-LDL+si-LTBP2+VEGF组细胞凋亡率和ROS含量均增加,细胞活力、SOD活性均降低(均为P<0.05)。结论LTBP2通过激活P38 MAPK信号通路促进VEGF表达加剧ox-LDL引起的ARPE-19细胞氧化应激损伤。Objective To explore the effects of latent TGF-βbinding protein 2(LTBP2)on oxidative injury and apoptosis in oxidized low-density lipoproteinox(ox-LDL)-treated human retinal pigment epithelial cell line(ARPE-19)cells.Methods The ARPE-19 cells was incubated with 0 mg·L^(-1),50 mg·L^(-1),100 mg·L^(-1) and 200 mg·L^(-1) ox-LDL,and cell viability and apoptosis were detected using CCK-8 kit and cell apoptosis detecting kit,respectively.The protein expression of LTBP2 in ARPE-19 cells after treated by different concentration ox-LDL was detected by Western blot.ARPE-19 cells incubated with 100 mg·L^(-1) ox-LDL were selected to build the oxidative damage model.ARPE-19 cells were randomly divided into control group,ox-LDL group,ox-LDL+ov-LTBP2(cells were transfected with overexpression vectors of LTBP2),ox-LDL+ov-NC(cells were transfected with negative controls,ox-LDL+si-LTBP2(cells were transfected with LTBP2 siRNA),ox-LDL+si-NC(cells were transfected with negative control siRNA)and ox-LDL+si-LTBP2+vascular endothelial growth factor(VEGF)group.After treatment,cell viability and apoptosis were detected through CCK-8 kit and cell apoptosis detecting kit,respectively.Superoxide dismutase(SOD),reactive oxygen species(ROS),and VEGF levels in culture medium were detected by ELISA assay,and protein expression of LTBP2,P38 and p-P38 were detected by Western blot.Results Compared with APRE-19 cells treated with 0 mg·L^(-1) ox-LDL,the cell viability of APRE-19 decreased,the apoptosis rate increased,and the level of LTBP2 protein e expression increased after 50 mg·L^(-1),100 mg·L^(-1) and 200 mg·L^(-1) ox-LDL treatment,and the changes were dose-dependent(all P<0.05).Compared with ox-LDL+ov-NC group,ox-LDL+ov-LTBP2 treatment up-regulated LTBP protein expression,raised cell apoptosis rate and ROS content,while decreased cell viability and SOD activity,but increased p-P38/P38 and the level of VEGF expression in APRE-19 cells(all P<0.05).Compared with the ox-LDL+si-NC group,the LTBP2 protein expression level,ROS content and a

关 键 词：潜伏转化生长因子β结合蛋白2(LTBP2) 氧化应激 视网膜色素上皮细胞 氧化型低密度脂蛋白 

分 类 号：R774[医药卫生—眼科]