Preliminary discussion on the potential mechanism of follistatin-like protein 1 in the process of proliferative diabetic retinopathy

Q4 Medicine

中华眼底病杂志 Pub Date : 2020-03-25 DOI:10.3760/CMA.J.CN511434-20190115-00024

Rui Niu, Lijie Dong, Xueli Du, Yanhua He, Zetong Nie, Weina Cui, Qiong Chen

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引用次数: 0

Abstract

Objective To observe the changes of follistatin-like protein 1 (FSTL1) in serum of patients with proliferative diabetic retinopathy (PDR). Methods Twenty PDR patients confirmed by clinical examination and 20 normal people were included in the study. Human retinal vascular endothelial cells (HRCEC) were divided into HRCEC blank control group, 3 h hypoxia group, 6 h hypoxia group. Human umbilical vein endothelial cell (HUVEC) were divided into HUVEC blank control group, 3h hypoxia group, 6h hypoxia group. Real-time quantitative PCR (RT-PCR) and ELISA were used to determine the expression of FSTL1, TGF-β, VEGF, connective tissue growth factor (CTGF) mRNA and protein in peripheral blood and cells of all groups from all subjects. Results The expressions of FSTL1, TGF-β1, CTGF, VEGF mRNA in blood samples of patients with PDR were 1.79±0.58, 0.97±0.21, 1.85±0.69 and 1.38±0.44. The expressions of FSTL1, TGF-β1 protein were 1.19±0.50, 0.71±0.24 ng/ml and 734.03±116.45, 649.36±44.23 ng/L. Compared with normal people, the differences were statistically significant (tmRNA=0.90, 0.21, 2.85, 1.77; P=0.00, 0.00, 0.04, 0.02. tprotein=1.88, 7.68; P=0.00, 0.02). The cell viability of HRCEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.66±0.05 and 0.64±0.04, respectively. Compared with the blank control group, the difference was statistically significant (F=13.02, P=0.00). The cell viability of HUVEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.63±0.06 and 0.68±0.06, respectively. Compared with the blank control group, the difference was statistically significant (F=26.52, P=0.00). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HRCEC blank control group and 3 h hypoxia group, the differences were statistically significant (F=14.75, 44.93, 85.54, 6.23; P=0.01, 0.00, 0.00, 0.03). Compared with the HRCEC blank control and 3 h hypoxia group, the expressions of FSTL1 and TGF-β1 protein were statistically significant (P<0.05). There was a statistically significant difference in TGF-β1 protein expression in the hypoxic 6 h group (P=0.03) and no significant difference in FSTL1 protein expression (P=0.68). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HUVEC blank control group and 3h hypoxia group, the differences were statistically significant (F=19.08, 25.12, 22.89, 13.07; P=0.00, 0.00, 0.00, 0.01). Immunofluorescence staining results showed that FSTL1, TGF-β1, CTGF, and VEGF proteins were positively expressed in cells in the 3h hypoxia and 6h hypoxia groups. Conclusion The expression of FSTL1 gene and protein in serum of PDR patients was significantly higher than that of normal people. Key words: Follistatin-related proteins; Human umbilical vein endothelial cells; Diabetic retinopathy

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卵泡抑素样蛋白1在增殖性糖尿病视网膜病变过程中的潜在机制初探

目的观察增生性糖尿病视网膜病变(PDR)患者血清卵泡他汀样蛋白1 (FSTL1)的变化。方法选取经临床检查证实的PDR患者20例，正常人20例。将人视网膜血管内皮细胞(HRCEC)分为HRCEC空白对照组、缺氧3 h组、缺氧6 h组。将人脐静脉内皮细胞(HUVEC)分为HUVEC空白对照组、缺氧3h组、缺氧6h组。采用实时荧光定量PCR (RT-PCR)和酶联免疫吸附法(ELISA)检测各组外周血和细胞中FSTL1、TGF-β、VEGF、结缔组织生长因子(CTGF) mRNA和蛋白的表达。结果PDR患者外周血FSTL1、TGF-β1、CTGF、VEGF mRNA表达量分别为1.79±0.58、0.97±0.21、1.85±0.69、1.38±0.44。FSTL1、TGF-β1蛋白表达量分别为1.19±0.50、0.71±0.24 ng/ml和734.03±116.45、649.36±44.23 ng/L。与正常人比较，差异有统计学意义(tmRNA=0.90, 0.21, 2.85, 1.77;P=0.00, 0.00, 0.04, 0.02。tprotein = 1.88, 7.68;P = 0.00, 0.02)。缺氧3 h和缺氧6 h时HRCEC细胞活力分别为0.66±0.05和0.64±0.04。与空白对照组比较，差异有统计学意义(F=13.02, P=0.00)。缺氧3 h组和缺氧6 h组HUVEC细胞活力分别为0.63±0.06和0.68±0.06。与空白对照组比较，差异有统计学意义(F=26.52, P=0.00)。HRCEC空白对照组与缺氧3 h组FSTL1、TGF-β1、CTGF、VEGF mRNA表达比较，差异均有统计学意义(F=14.75、44.93、85.54、6.23;P=0.01, 0.00, 0.00, 0.03)。与HRCEC空白对照组和缺氧3 h组比较，FSTL1、TGF-β1蛋白表达均有统计学意义(P<0.05)。缺氧6 h组TGF-β1蛋白表达差异有统计学意义(P=0.03)， FSTL1蛋白表达差异无统计学意义(P=0.68)。HUVEC空白对照组与缺氧3h组FSTL1、TGF-β1、CTGF、VEGF mRNA表达比较，差异均有统计学意义(F=19.08、25.12、22.89、13.07;P=0.00, 0.00, 0.00, 0.01)。免疫荧光染色结果显示，缺氧3h和缺氧6h组细胞中FSTL1、TGF-β1、CTGF、VEGF蛋白均阳性表达。结论PDR患者血清FSTL1基因及蛋白表达明显高于正常人。关键词:卵泡抑素相关蛋白;人脐静脉内皮细胞;糖尿病性视网膜病变

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来源期刊

中华眼底病杂志 Medicine-Ophthalmology

CiteScore

0.40

自引率

0.00%

发文量

5383

期刊介绍： Chinese Journal of Ocular Fundus Diseases is the only scientific journal in my country that focuses on reporting fundus diseases. Its purpose is to combine clinical and basic research, and to give equal importance to improvement and popularization. It comprehensively reflects the leading clinical and basic research results of fundus disease disciplines in my country; cultivates professional talents in fundus disease, promotes the development of fundus disease disciplines in my country; and promotes academic exchanges on fundus disease at home and abroad. The coverage includes clinical and basic research results of posterior segment diseases such as retina, uveal tract, vitreous body, visual pathway, and internal eye diseases related to systemic diseases. The readers are medical workers and researchers related to clinical and basic research of fundus diseases. According to the journal retrieval report of the Chinese Institute of Scientific and Technological Information, the comprehensive ranking impact factor and total citation frequency of the Chinese Journal of Ocular Fundus Diseases have been among the best in the disciplines of ophthalmology, otolaryngology, and ophthalmology in my country for many years. The papers published have been included in many important databases at home and abroad, such as Scopus, Peking University Core, and China Science Citation Database (CSCD).