摘要目的:观察增生型糖尿病视网膜病变（PDR）患者血清中卵泡抑素样蛋白1 （FSTL1）的变化。方法:临床检查确诊的PDR患者（PDR组）20例及正常人20名（正常组）纳入研究。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；人脐静脉内皮细胞（HUVEC）分为HUVEC空白对照组、缺氧3 h组、缺氧6 h组。应用实时定量PCR （RT-PCR）及ELISA测定所有受检者外周血和各组细胞中FSTL1、TGF-β、VEGF、结缔组织生长因子（CTGF）mRNA和蛋白表达。受检者血清中FSTL1、TGF-β1、CTGF、VEGF mRNA和蛋白比较行独立样本 t检验；HRCEC空白对照组、缺氧3 h组、缺氧6 h组，HUVEC空白对照组、缺氧3 h组、缺氧6 h组细胞生存力、mRNA表达比较行单因素方差分析。 结果:PDR组患者血清中FSTL1、TGF-β1、CTGF、VEGF mRNA表达分别为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。与正常组比较，差异均有统计学意义（ tmRNA=0.90、0.21、2.85、1.77 ， P=0.00、0.00、0.04、0.02 ； t蛋白＝1.88、7.68， P=0.00、0.02）。HRCEC缺氧3 h组、缺氧6 h组细胞生存力分别为0.66±0.05、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）。HUVEC缺氧3 h组、缺氧6 h组细胞生存力分别为0.63±0.06、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、缺氧3 h组细胞中FSTL1、TGF-β1、CTGF、VEGF mRNA表达比较，差异有统计学意义（ F=14.75、44.93、85.54、6.23， P=0.01、0.00、0.00、0.03 ）；与HRCEC空白对照组上清液中FSTL1、TGF-β1蛋白表达比较，缺氧3 h组，蛋白表达差异均有统计学意义（ 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、缺氧3 h组、缺氧6 h组细胞中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3 h组、缺氧6 h组细胞中FSTL1、TGF-β1、CTGF、VEGF蛋白均呈阳性表达。 结论:PDR患者血清中FSTL1基因及蛋白表达较正常人显著升高。

abstractsObjective: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.