【摘要】 目的探讨骨形态发生蛋白-6（BMP-6）对转化生长因子-β2（TGF-β2）所致视网膜色素上皮（RPE）细胞迁移的影响。方法 RPE细胞常规培养后,用5μg·L^-1 TGF-β2处理48 h, Western blot检测细胞中BMP-6蛋白水平的变化,Transwell法检测细胞迁移能力变化。随后,筛选BMP-6 siRNA抑制率最高的片段用于后续实验,构建BMP-6过表达质粒并进行鉴定后用于后续实验,RPE细胞随机分组,即（1）对照组:加入正常培养基培养48 h;（2）TGF-β2组:培养基中加入5μg·L^-1 TGF-β2培养48 h;（3）BMP-6 siRNA组:转染BMP-6-homo-1087后培养48 h;（4）BMP-6过表达组:转染BMP-6过表达质粒后培养48 h;（5）TGF-β2+BMP-6过表达组:转染BMP-6过表达质粒24 h后,加入5μg·L^-1TGF-β2继续培养48 h。Transwell法检测细胞迁移数的变化,倒置显微镜观察细胞形态的变化。结果 TGF-β2组迁移细胞数为（122.00±5.57）个,与对照组[（63.67±4.04）个]相比差异有统计学意义（P=0.000）。TGF-β2组BMP-6蛋白表达水平为0.41±0.13,与对照组（0.70±0.08）相比差异具有统计学意义（P=0.031）。BMP-6 siRNA组迁移细胞数为（115.67±1.53）个,与对照组[（57.00±7.00）个]相比差异具有统计学意义（P=0.000）。BMP-6过表达组迁移细胞数为（44.33±5.51）个,与对照组[（60.00±6.25）个]相比差异无统计学意义（P=0.076）。TGF-β2+BMP-6过表达组迁移细胞数为（90.33±3.51）个,与TGF-β2组[（112.00±9.64）个]相比差异具有统计学意义（P=0.016）。结论 BMP-6可以阻止TGF-β2所致的RPE细胞的迁移,为增生性玻璃体视网膜病变的防治提供新的视角。更多还原

【Abstract】 Objective To investigate the effect of bone morphogenetic protein-6（BMP-6） on the migration of retinal pigment epithelial（RPE） cells induced by the transforming growth factor-β2（TGF-β2）.Methods After being routinely cultured, RPE cells were treated with 5 μg·L^-1 TGF-β2 for 48 h. Their BMP-6 protein levels were detected by the Western blot, and cell migration was detected by the Transwell chamber. Subsequently, the fragment with the highest inhibition rate of BMP-6 siRNA was selected for subsequent experiments, and BMP-6 overexpression plasmids were constructed and identified for use in subsequent experiments. RPE cells were randomly divided into（1）control group: cultured with the normal medium for 48 h;（2）TGF-β2 group: cultured with 5 μg·L^-1 TGF-β2 for 48 h;（3）BMP-6 siRNA group: transfected with the BMP6-homo-1087 and cultured for 48 h;（4）BMP-6 overexpression group: transfected with the BMP-6 overexpression plasmid and cultured for 48 h;（5）TGF-β2+BMP-6 overexpression group: after transfection with the BMP-6 overexpression plasmid for 24 h, cultured for 48 h with additional 5 μg·L^-1 TGF-β2. The Transwell chamber was used to detect the change in the number of migrating cells, and the inverted microscope to observe the change in the cell morphology.Results The number of migrating cells in the TGF-β2 group was 122.00±5.57, which was significantly higher than that in the control group（63.67±4.04）（P=0.000）. The expression level of BMP-6 protein in the TGF-β2 group was 0.41±0.13, which was significantly lower than that in the control group（0.70±0.08）（P=0.031）. The number of migrating cells in the BMP-siRNA group was 115.67±1.53, which was significantly higher than that in the control group（57.00±7.00）（P=0.000）. The number of migrating cells in the BMP-6 overexpression group was 44.33±5.51, which had significant no difference when compared with the control group（60.00±6.25）（P=0.076）. The number of migrating cells in the TGF-β2+BMP-6 overexpression group was 90.33±3.51, which was significantly lower than that in the TGF-β2 group（112.00±9.64）（P=0.016）.Conclusion BMP-6 can prevent the migration of RPE cells caused by TGF-β2, which may provide a new insight for the prevention and treatment of proliferative vitreoretinopathy.更多还原