SPC对人真皮成纤维细胞中MMP-1、MMP-2、MMP-3表达影响的研究

发布时间：2018-06-28 03:54

本文选题：神经鞘氨醇磷酸胆碱 + 成纤维细胞　；参考：《大连医科大学》2011年硕士论文

【摘要】：目的:本实验采用人真皮成纤维细胞作为实验模型,研究神经鞘氨醇磷酸胆碱(sphingosylphosphorylchoine, SPC)对人真皮成纤维细胞中基质金属蛋白酶-1(MMP-1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-3(MMP-3)表达的影响,探讨SPC促进皮肤创伤愈合作用机制。方法:采用组织块消化法培养人真皮成纤维细胞,取4-10代进行实验,利用逆转录聚合酶链反应(RT-PCR)和免疫印迹(Western blotting)的方法半定量检测经SPC诱导的人真皮成纤维细胞中MMP-1、MMP-2、MMP-3 mRNA水平和蛋白水平的表达。结果:采用组织块消化法培养人真皮成纤维细胞,与传统的消化法和组织法比较,成功率较高。SPC以时间梯度(2、6、12、24和48小时)诱导人真皮成纤维细胞后,RT-PCR结果显示,mRNA的表达水平产生了不同程度的变化,MMP-1、MMP-3在SPC作用2小时左右时逐渐升高,六小时左右达到高峰,维持一段时间后下降。MMP-2 mRNA的表达水平变化不显著。Western blotting分析显示,SPC对MMP-1、MMP-3蛋白水平的影响同样具有时间依赖性,MMP-2蛋白表达变化不明显。结论:SPC促进人真皮成纤维细胞MMP-1、MMP-3的表达,进而参与调节细胞外基质,可能为SPC促进皮肤创伤愈合的作用机制之一。
[Abstract]:Aim: to study the effects of sphingosylphosphorylchoine (SPC) on the expression of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-3 (MMP-3) in human dermal fibroblasts. To explore the mechanism of SPC promoting skin wound healing. Methods: human dermal fibroblasts were cultured by tissue mass digestion and cultured for 4-10 passages. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting (Western blotting) were used to detect the expression of MMP-3 mRNA and protein in human dermal fibroblasts. Results: human dermal fibroblasts were cultured by tissue mass digestion, and compared with the traditional digestion method and organic law. The results of RT-PCR showed that the level of mRNA expression of MMP 1 + MMP-3 increased gradually after SPC was exposed to SPC for 2 hours or so, and reached its peak at about 6 hours after inducing human dermal fibroblasts with a time gradient of 2 h and 48 h, the results showed that the mRNA expression level of MMP 1 + MMP-3 increased gradually at about 2 h after SPC treatment, and reached its peak at about 6 h. Western blotting analysis showed that the effect of SPC on the expression of MMP-2 protein was also time-dependent, and there was no significant change in the expression of MMP-2 protein. Conclusion SPC can promote the expression of MMP-1 and MMP-3 in human dermal fibroblasts, and then regulate the extracellular matrix, which may be one of the mechanisms of promoting skin wound healing by SPC.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R751

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