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目的:观察氧化应激条件下骨形成蛋白4(BMP4)对人视网膜微血管内皮细胞(hRMEC)的增生和迁移影响。方法:将体外培养的hRMEC分为对照组、4-羟基壬烯醛(HNE)处理组(4-HNE组)、4-HNE+BMP4处理组(BMP4组)。4-HNE组细胞培养基加入10 μmmol/L 4-HNE;BMP4组细胞培养基加入10 μmmol/L 4-HNE刺激6 h后,再加入100 ng/ml重组人BMP4。噻唑蓝比色法检测4-HNE、BMP4对hRMEC生存力的影响。细胞划痕实验测定4-HNE、BMP4对细胞迁移能力的影响。实时定量聚合酶链反应(qRT-PCR)检测对照组、4-HNE组细胞中BMP4 mRNA相对表达量以及对照组、BMP4组细胞中纤维连接蛋白(FN)、层粘连蛋白(Laminin)、α-平滑肌收缩蛋白(α-SMA)、胶原蛋白Ⅰ型(Collagen Ⅰ)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)mRNA相对表达量。蛋白免疫印迹法(Western blot)检测对照组、4-HNE组细胞中BMP4蛋白相对表达量。两组间比较采用n t检验。n 结果:与对照组比较,4-HNE组、BMP4组细胞生存力(n t=12.73、16.26,n P=0.000 2、<0.000 1)、细胞迁移率(n t=28.17、37.48,n P<0.000 1、<0.000 1)均明显提高,差异有统计学意义;4-HNE组细胞中BMP4 mRNA、蛋白相对表达量明显升高,差异有统计学意义(n t=16.36、69.35,n P=0.000 1、<0.000 1)。qRT-PCR检测结果显示,与对照组比较,BMP4组细胞中VEGF、FN、Laminin、α-SMA、Collagen Ⅰ、CTGF mRNA相对表达量明显升高,差异有统计学意义(n t=10.61、17.00、14.85、7.78、12.02、10.61,n P=0.0004、<0.000 1、0.000 1、0.001 5、0.000 1、0.000 4)。n 结论:BMP4可诱导hRMEC的增生和迁移,并可调控hRMEC中血管生成因子和纤维化相关因子表达。“,”Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by n t test.n Results:Compared with the control group, cell viability (n t=12.73, 16.26, n P=0.000 2, <0.000 1), cell migration rate ( n t=28.17, 37.48, n P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant (n t=16.36, 69.35, n P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( n t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, n P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4).n Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.