摘要
目的探讨转甲状腺素蛋白(transthyretin,TTR)在糖尿病视网膜病变(diabetic retinopathy,DR)患者中的表达及对高糖(high glucose,HG)诱导的人视网膜色素上皮细胞(human retinal pigment epithelium cells 19,ARPE-19)增殖、迁移和血管形成的影响和机制。方法收集DR患者(n=43)和健康体检者(n=45)血清,采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清TTR和血管内皮生长因子(vascular endothelial growth factor,VEGF)水平,逆转录-定量聚合酶链反应(reverse transcription-quantitative polymerase chain reaction,RT-PCR)检测血清中长链非编码RNA母系表达基因3(maternally expressed gene 3,MEG3)水平。将细胞分为对照组、HG组、HG+TTR组、HG+pcDNA3.1组、HG+pcDNA3.1-MEG3组、HG+TTR+pcDNA3.1-MEG3组,qRT-PCR和Western blot检测各组细胞中TTR、MEG3和VEGF表达水平,细胞计数试剂(cell counting kit-8,CCK8)检测细胞增殖,transwell试验检测细胞迁移,BD Matrigel基质培养测定血管形成。结果与健康体检者比较,DR患者血清TTR和VEGF水平显著升高,MEG3水平显著降低(t值分别为7.81,13.04,11.89,P均<0.001)。与对照组比较,HG组TTR和VEGF蛋白表达升高,MEG3的mRNA水平降低(t值分别为6.45,5.68,8.13,P均<0.05)。与HG组比较,HG+TTR组VEGF蛋白表达升高,MEG3的mRNA水平降低(t值分别为7.43,9.29,P均<0.01),细胞的增殖、迁移和血管形成减少(t值分别为5.32,3.22,4.32,P<0.01),而HG+pcDNA3.1-MEG3组TTR表达无差异(P>0.05),MEG3表达上调(t=12.02,P<0.01),VEGF表达下调(t=7.05,P<0.01),促进了细胞增殖、迁移和血管形成(t值分别为6.45,5.22,11.32,P均<0.01)。结论TTR在DR患者血清和HG诱导的ARPE-19细胞中高表达,可能通过调控lncRNA MEG3表达上调VEGF表达,抑制ARPE-19细胞增殖、迁移和血管形成。
Objective To explore the expression of transthyretin(TTR)in patients with diabetic retinopathy(DR)and its effect mechanism on the proliferation,migration and angiogenesis of human retinal pigment epithelial cells 19(ARPE-19)induced by high glucose(HG).Methods The serum levels of DR patients(n=43)and healthy subjects(n=45)were collected,the serum levels of TTR and vascular endothelial growth factor(VEGF)were detected by enzyme linked immunosorbent assay(ELISA),the level of long-chain non-coding RNA maternally expressed gene 3(MEG3)in serum was detected by reverse transcription-quantitative polymerase chain reaction(RT-PCR).The cells were divided into control group,HG group,HG+TTR group,HG+pcDNA3.1 group,HG+pcDNA3.1-MEG3 group,HG+TTR+pcDNA3.1-MEG3 group,the expression levels of TTR,MEG3 and VEGF in each group were detected by RT-PCR and Western blot,the proliferation of cell was detected by cell counting kit-8(CCK8),the migration of cell was detected by transwell test,and the angiogenesis of cell was measured by BD Matrigel matrix culture.Results Compared with healthy subjects,serum TTR and VEGF levels of DR patients were significantly increased,and MEG3 levels were significantly reduced(t values were 7.81,13.04,11.89,respectively,all P<0.001).Compared with the control group,the expression of TTR and VEGF protein in the HG group were increased,and the mRNA level of MEG3 were decreased(t values were 6.45,5.68,8.13,respectively,all P<0.05).Compared with the HG group,the VEGF protein expression in the HG+TTR group was increased,the mRNA level of MEG3 was decreased(t values were 7.43,9.29,all P<0.01),the proliferation,migration and angiogenesis of cells in the HG+TTR group were reduced(t values were 5.32,3.22,4.32,respectively,P<0.01).There was no significantly difference of TTR expression between the HG+pcDNA3.1-MEG3 group and HG group,while the MEG3 expression in the HG+pcDNA3.1-MEG3 group was up-regulated(t=12.02,P<0.01),the VEGF expression was down-regulated(t=7.05,P<0.01),the proliferation,migration and angiogenes
作者
齐新刚
顾永欣
肖云
杜雷
QI Xingang;GU Yongxin;XIAO Yun;DU Lei(Department of Army Ophthalmology Center,Beijing Road Medical District of General Hospital of Xinjiang Military Command,Urumqi Xinjiang830013,China)
出处
《华南国防医学杂志》
CAS
2020年第11期757-761,776,共6页
Military Medical Journal of South China
基金
新疆维吾尔自治区自然科学基金项目(2019D01C410)。
