视网膜色素上皮细胞LncRNA MEG3在不同葡萄糖浓度下的表达情况及对VEGF和TGF-β1表达的影响  

Expression of long non-coding RNA maternally expressed gene 3 in retinal pigment epithelial cells under different glucose concentrations and its effect on vascular endothelial growth factor and transforming growth factorβ1

作　　者：张达宁[1] 冷瀛[1] 秦海翔[1] 李相军[1] 白丹[1] 石盛玲 王敬怡 卢奕安 张富程 陈艳 ZHANG Daning;LENG Ying;QIN Haixiang;LI Xiangjun;BAI Dan;SHI Shengling;WANG Jingyi;LU Yian;ZHANG Fucheng;CHEN Yan(Department of Ophthalmology,Affiliated Hospital of Beihua University,Jilin 132011,Jilin Province,China)

机构地区：[1]北华大学附属医院眼科,吉林省吉林市132011

出　　处：《眼科新进展》2022年第7期524-528,共5页Recent Advances in Ophthalmology

基　　金：吉林省卫生健康技术创新项目(编号:2019J048)。

摘　　要：目的探讨长链非编码RNA母系表达基因3(LncRNA MEG3)在不同浓度葡萄糖下表达情况及对血管内皮生长因子(VEGF)、转化生长因子β1(TGF-β1)表达的影响。方法按照不同浓度d-葡萄糖培养ARPE-19细胞,随机分为5 mmol·L^(-1)d-葡萄糖组、15 mmol·L^(-1)d-葡萄糖组、30 mmol·L^(-1)d-葡萄糖组。实时荧光定量PCR(RT-qPCR)检测各组ARPE-19细胞LncRNA MEG3、VEGF mRNA、TGF-β1 mRNA相对表达水平。ARPE-19细胞转染PCDNA-MEG3及PCDNA-NC质粒,建立LncRNA MEG3过表达ARPE-19细胞及转染空质粒的ARPE-19细胞。以含30 mmol·L^(-1)d-葡萄糖的DMEM分别培养以下各组ARPE-19细胞:LncRNA MEG3过表达ARPE-19细胞(O组);转染空质粒的ARPE-19细胞(NC组);加入PBS的正常ARPE-19细胞(G组);同时以含5 mmol·L^(-1)d-葡萄糖的DMEM培养正常ARPE-19细胞(C组);以上各组细胞培养24 h后,采用RT-qPCR法及Western blot法检测各组ARPE-19细胞VEGF mRNA和VEGF蛋白、TGF-β1 mRNA和TGF-β1蛋白的相对表达水平。结果与5 mmol·L^(-1)d-葡萄糖组比较,15 mmol·L^(-1)d-葡萄糖组和30 mmol·L^(-1)d-葡萄糖组ARPE-19细胞LncRNA MEG3相对表达水平降低,VEGF mRNA、TGF-β1 mRNA相对表达水平升高,差异均有统计学意义(均为P<0.05)。与15 mmol·L^(-1)d-葡萄糖组比较,30 mmol·L^(-1)d-葡萄糖组ARPE-19细胞LncRNA MEG3相对表达水平降低,VEGF mRNA、TGF-β1 mRNA相对表达水平升高,差异均有统计学意义(均为P<0.05)。与G组和NC组比较,O组ARPE-19细胞VEGF mRNA和VEGF蛋白、TGF-β1 mRNA和TGF-β1蛋白相对表达水平均降低,差异均有统计学意义(均为P<0.05)。结论高糖下调ARPE-19细胞LncRNA MEG3的表达,上调VEGF和TGF-β1的表达。LncRNA MEG3过表达降低高糖诱导的VEGF和TGF-β1表达的升高。LncRNA MEG3可能通过抑制VEGF和TGF-β1的表达调节糖尿病视网膜病变的发展。Objective To investigate the expression of long non-coding RNA maternally expressed gene 3(LncRNA MEG3)under different glucose concentrations and its effect on the expression of vascular endothelial growth factor(VEGF)and transforming growth factorβ1(TGF-β1).Methods ARPE-19 cells cultured in media with different d-glucose concentrations were randomly divided into 5 mmol·L^(-1) d-glucose group,15 mmol·L^(-1) d-glucose group,and 30 mmol·L^(-1) d-glucose group.The relative expression levels of LncRNA MEG3,VEGF mRNA,and TGF-β1 mRNA in each group were detected by real-time quantitative polymerase chain reaction(RT-qPCR).ARPE-19 cells were transfected with PCDNA-MEG3 and PCDNA-NC plasmids to establish LncRNA MEG3 overexpressed ARPE-19 cells and empty plasmid transfected ARPE-19 cells.LncRNA MEG3 overexpressed ARPE-19 cells cultured in DMEM containing 30 mmol·L^(-1) d-glucose were set as group O.Empty plasmid transfected ARPE-19 cells cultured in DMEM containing 30 mmol·L^(-1) d-glucose were set as group NC.Normal ARPE-19 cells added with PBS cultured in DMEM containing 30 mmol·L^(-1) d-glucose were set as group G.Normal ARPE-19 cells cultured in DMEM containing 5 mmol·L^(-1) d-glucose were set as group C.After the cells were incubated for 24 h,the relative expression levels of VEGF mRNA and VEGF protein,TGF-β1 mRNA and TGF-β1 protein in each group were detected by RT-qPCR and Western blot.Results Compared with the 5 mmol·L^(-1) d-glucose group,the relative expression level of LncRNA MEG3 in the 15 mmol·L^(-1) d-glucose and 30 mmol·L^(-1) d-glucose groups was decreased,while the relative expression levels of VEGF mRNA and TGF-β1 mRNA were increased(all P<0.05).Compared with the 15 mmol·L^(-1) d-glucose group,the relative expression level of LncRNA MEG3 in the 30 mmol·L^(-1) d-glucose group was decreased,while the relative expression levels of VEGF mRNA and TGF-β1 mRNA were increased(all P<0.05).Compared with group G and group NC,the relative expression levels of VEGF mRNA and VEGF protein,TGF-β1 mRNA

关 键 词：长链非编码RNA母系表达基因3 血管内皮生长因子 转化生长因子Β1 视网膜色素上皮细胞 

分 类 号：R774.1[医药卫生—眼科]