T淋巴细胞表达CD147对单核细胞趋化和分泌MMP-9的影响

发布时间：2017-12-31 07:45

本文关键词：T淋巴细胞表达CD147对单核细胞趋化和分泌MMP-9的影响　出处：《蚌埠医学院》2011年硕士论文　论文类型：学位论文

【摘要】：背景：慢性炎症反应能导致动脉血管壁的损伤和内皮功能的紊乱，T淋巴细胞和单核巨噬细胞作为重要的炎症细胞，在动脉粥样硬化发生发展的过程中起到了重要作用。T淋巴细胞高表达的CD147是否对单核细胞趋化性和MMPs的分泌有影响，CypA是否通过T细胞的表达的CD147而促进单核细胞趋化及MMPs分泌，以及CsA是否对此有抑制作用，有待进一步研究。目的：验证T淋巴细胞表达的CD147对单核细胞趋化和分泌MMP-9的影响，亲环素A是否通过Jurkat细胞表达的CD147影响单核细胞趋化和MMP-9的表达，以及CsA是否对此有抑制作用，从而揭示动脉粥样硬化在发生和发展中可能涉及的免疫机制，为防治动脉粥样硬化提供新的有效的靶点。方法：体外培养人Jurkat T淋巴细胞株，转染针对CD147特异的siRNA抑制CD147在Jurkat T淋巴细胞的表达，分别从基因水平和蛋白水平验证siRNA对CD147的抑制效率。建立体外人Jurkat T淋巴细胞和THP-1单核细胞共培养体系，在共培养前分别用亲环素A和环孢素A预处理Jurkat细胞，转染siRNA抑制CD147在Jurkat细胞的表达，亲环素A预处理siRNA抑制CD147表达后的Jurkat细胞，，环孢素A预处理后再加入亲环素A的Jurkat细胞；各共培养体系中分别共培养48h后收集培养液上清，用ELISA检测各组中单核细胞分泌MMP-9量的变化；用Millicell趋化小室，做趋化实验检测上述各组共培养体系中不同干预条件下对单核细胞趋化性的影响。结果：针对CD147的siRNA能特异性的抑制Jurkat T淋巴细胞CD147的表达，与对照组相比siRNA干扰组细胞表面CD147的平均荧光强度（MFI）下降（50.5±4.7）%（p0.05），CD147mRNA和蛋白水平的表达均降低，抑制率分别为（38.57±1.55）%、（47.4±1.47）%（p0.05），符合实验要求。与正常组比较，siRNA特异性抑制CD147表达的Jurkat细胞与THP-1细胞共培养单核细胞分泌MMP-9量和趋化能力均明显降低（p0.05）；亲环素A刺激后的Jurkat细胞与单核细胞共培养体系中单核细胞分泌MMP-9量和趋化能力均明显增加（p0.05），而加入环孢素A预处理的Jurkat细胞与THP-1单核细胞共培养，单核细胞分泌MMP-9量和趋化能力均降低（p0.05）；分别用亲环素A刺激siRNA抑制CD147后的Jurkat T淋巴细胞和环孢素A预处理后的Jurkat T淋巴细胞后再与THP-1单核细胞共培养，在以上两个共培养体系中单核细胞趋化和分泌MMP-9的量变化不明显（p0.05），差异无统计学意义。结论：1．化学合成的siRNA能特异性抑制Jurkat T淋巴细胞CD147基因和蛋白的表达，为下一步研究CD147在动脉粥样硬化中的作用奠定了基础。 2．体外成功建立Jurkat T淋巴细胞和THP-1单核细胞的共培养体系；抑制了CD147在Jurkat细胞上的表达可使共培养体系中单核细胞趋化性和MMP-9的分泌降低，说明T淋巴细胞表达的CD147可影响共培养体系中单核细胞趋化性和MMP-9表达的有效途径。 3．亲环素A可通过Jurkat细胞上表达的CD147对共培养体系中单核细胞趋化和MMP-9的分泌起作用，而环孢素A可阻断亲环素A的这一效应。
[Abstract]:Background : Chronic inflammatory reaction can lead to vascular wall injury and endothelial dysfunction , T lymphocytes and mononuclear phagocytes play an important role in the development of atherosclerosis . Objective : To verify the effect of CD147 expressed by T lymphocytes on monocyte chemoattractant and MMP - 9 secretion , and whether cyclophilin A has an inhibitory effect on monocyte chemoattractant and MMP - 9 expression via CD147 expressed in the cells , as well as whether CsA has an inhibitory effect on the expression of MMP - 9 and whether CsA can be involved in the pathogenesis and development of atherosclerosis , which can provide a new effective target for preventing and treating atherosclerosis . Methods : The CD147 - specific siRNA was cultured in vitro to inhibit CD147 expression . After co - culture , the cells were pretreated with cyclosporins A and cyclosporine A . After 48 hours of co - culture , the expression of MMP - 9 was inhibited and the expression of MMP - 9 was inhibited in each group . The effects of different intervention conditions on monocyte chemoattractant were detected by ELISA . Results : Compared with the control group , the expression of MMP - 9 and MCP - 9 decreased significantly ( P < 0.05 ) . Compared with the control group , the expression of MMP - 9 and MCP - 9 decreased significantly ( P < 0.05 ) . Compared with the control group , the amount of MMP - 9 and monocyte - secreting MMP - 9 were significantly decreased ( P < 0.05 ) . Conclusion : 1 . The chemically synthesized siRNA can specifically inhibit the expression of CD147 gene and protein , and lays a foundation for the next study on the role of CD147 in atherosclerosis . 2 . The co - culture system of T lymphocytes and THP - 1 monocytes was successfully established in vitro , and the expression of CD147 was inhibited in the co - culture system and the secretion of MMP - 9 in the co - culture system was decreased . 3 . Cyclophilin A can inhibit the secretion of monocyte chemoattractant and MMP - 9 in the co - culture system by CD147 expressed on the cells , and cyclosporin A can block the effect of cyclophilin A .

【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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