本文選題：神經(jīng)鞘氨醇磷酸膽堿 + 成纖維細胞�。� 參考：《大連醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:本實驗采用人真皮成纖維細胞作為實驗?zāi)Ｐ?研究神經(jīng)鞘氨醇磷酸膽堿(sphingosylphosphorylchoine, SPC)對人真皮成纖維細胞中基質(zhì)金屬蛋白酶-1(MMP-1)、基質(zhì)金屬蛋白酶-2(MMP-2)、基質(zhì)金屬蛋白酶-3(MMP-3)表達的影響,探討SPC促進皮膚創(chuàng)傷愈合作用機制。 方法:采用組織塊消化法培養(yǎng)人真皮成纖維細胞,取4-10代進行實驗,利用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)和免疫印跡(Western blotting)的方法半定量檢測經(jīng)SPC誘導(dǎo)的人真皮成纖維細胞中MMP-1、MMP-2、MMP-3 mRNA水平和蛋白水平的表達。 結(jié)果:采用組織塊消化法培養(yǎng)人真皮成纖維細胞,與傳統(tǒng)的消化法和組織法比較,成功率較高。SPC以時間梯度(2、6、12、24和48小時)誘導(dǎo)人真皮成纖維細胞后,RT-PCR結(jié)果顯示,mRNA的表達水平產(chǎn)生了不同程度的變化,MMP-1、MMP-3在SPC作用2小時左右時逐漸升高,六小時左右達到高峰,維持一段時間后下降。MMP-2 mRNA的表達水平變化不顯著。Western blotting分析顯示,SPC對MMP-1、MMP-3蛋白水平的影響同樣具有時間依賴性,MMP-2蛋白表達變化不明顯。 結(jié)論:SPC促進人真皮成纖維細胞MMP-1、MMP-3的表達,進而參與調(diào)節(jié)細胞外基質(zhì),可能為SPC促進皮膚創(chuàng)傷愈合的作用機制之一。
[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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R751

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