【摘要】 目的观察巨噬细胞条件培养液(MφCM)及白细胞介素-1β(IL-1β)对视网膜色素上皮(RPE)细胞分泌基质金属蛋白酶(MMP)-2及MMP-9的影响,探讨巨噬细胞与RPE细胞之间的相互关系。方法在人RPE细胞培养液中加入50%体积分数的人MφCM作用48h;加入终质量浓度为10、50、100、200ng/ml重组人IL-1β作用12、24、48h。用酶谱法检测RPE细胞培养液中MMP-2及MMP-9,凝胶图像扫描分析仪定量。结果MφCM促RPE细胞分泌MMP-2和MMP-9,促MMP-9分泌尤其明显;48h内IL-1β促RPE细胞分泌MMP-2和MMP-9具有时间和质量浓度依赖性。结论MφCM中含有促RPE细胞分泌MMP-2和MMP-9的因子,IL-1β是巨噬细胞产生的主要炎性因子,提示巨噬细胞可能通过分泌细胞因子调节RPE细胞分泌MMP-2和MMP-9,参与细胞增生、迁移等病理过程。更多还原

【Abstract】 ObjectiveRecent research showed that matrix metalloproteinase-2(MMP-2) and MMP-9 are overexpressed in subretinal fluid,vitreous fluid and preretianl membrane in proliferative vitreoretinopathy(PVR),but the cell type producing MMP-2 and MMP-9 is unclear.This study tried to investigate the effects of macrophages conditioned media(MφCM) and interleukin-1β(IL-1β) on the secretion of MMP-2 and MMP-9 in human retinal pigment epithelial(PRE) cells in vitro.MethodsThe RPE cells from fatal corpse eyes were cultivated in DMEM containing 15% FBS.DMEM containing 50% human peritoneal MφCM was added in culture for 48 hours and concentrations of IL-1β(0,10,50,100,200 ng/ml) were added in medium for 12,24,48 hours respectively in different groups.The cell culture supernatants was collected for MMP-2 and MMP-9 detect using zymography and the results were analyzed by densitometry.ResultsThe zymography of supernatants of RPE cells showed two clear white bands independently at 92 000 and 62 000 of protein molecular weight,indicating expression of MMP-2 and MMP-9.The expressions were significantly increased after the stimulation of MφCM in comparison with control group(P<0.05,P<0.05),and similar results were found after treatment by different concentrations of IL-1β(P<0.05).IL-1β increased MMP-2 and MMP-9 levels at dose- and time-dependent manner.ConclusionMφCM contains some factors promoting MMP-2 and MMP-9 secretion by RPE cells in vitro,and macrophages can regulate MMP-2 and MMP-9 expression by secreting certain cytokines.更多还原