溶血磷脂酸对人类视网膜色素上皮细胞生物学功能的影响研究

Effects of lysophosphatidic acid on the biological function of human retinal pigment epithelial cells in vitro

目的:研究溶血磷脂酸(lysophosphatidic acid,LPA)对体外培养人类视网膜色素上皮(human retinal pigment epithelia-lium,hRPE)细胞增殖活力、分化特性及免疫活性等生物学功能的影响。方法:采用MTT法检测体外培养hRPE细胞增殖活力;角蛋白及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)免疫细胞化学染色方法分析体外培养hRPE细胞分化特性;流式细胞术检测hRPE细胞表面HLA-ABC、HLA-DR、ICAM-1的表达。结果:加药后第2d开始LPA组吸光度明显高于对照组(P<0.001);角蛋白18表达对照组较LPA组强(uc=-12.683,P<0.001),α-SMA表达对照组较LPA组弱(uc=10.402,P<0.001);hRPE细胞表面HLA-ABC、HLA-DR、ICAM-1的阳性细胞率和平均荧光强度组间差异无统计学意义(各指标均P>0.05)。结论:LPA明显促进hRPE的增殖活力,促进hRPE转分化,在PVR形成过程中起潜在的促进作用,但对hRPE免疫活性无影响。

AIM：To investigate the effect of lysophosphatidic acid（LPA） on the cell proliferation,cell differentiation and immunological competence of human retinal pigment epithelialium（hRPE） cells in vitro METHODS：The cell proliferation of hRPE was observed by MTT.The cell differentiation of hRPE was observed by immunocytochemical staining to detect the expressions of keratin and α-smooth muscle actin.And the immunological competence of hRPE was observed by flow cytometry to detect the expressions of HLA-ABC,HLA-DR and ICAM-1.RESULTS：The absorbance of the LPA group was higher than that of the control group from the second day after administrating LPA（P〈0.001）.The expression of keratin 18 in the control group was stronger than that of the LPA group（P〈0.001）and the expression of α-smooth muscle actin in the control group was weaker than that of the LPA group（P〈0.001）.The differences of the positive cell rate and the mean fluorescence intensity of HLA-ABC,HLA-DR,ICAM-1between the LPA group and the control group had no statistical significance.（P〉0.05）.CONCLUSION：LPA promotes the cell proliferation of hRPE obviously.Furthermore,LPA promotes the cell transdifferentiation of hRPE,which plays a role in the development of proliferative vitreoretinal disorders.However,LPA dose not affect the immunological competence of hRPE.