【摘要】：目的:探討1-磷酸鞘氨醇(S1P)對ARPE-19細(xì)胞分泌炎性細(xì)胞因子的影響以及細(xì)胞信號傳導(dǎo)通路在其中的作用。 方法: ARPE-19細(xì)胞中S1P1-5受體的表達(dá)通過RT-PCR和realtime PCR進(jìn)行檢測。ARPE-19細(xì)胞經(jīng)TNF-α、S1P、百日咳毒素(Pertussiutoxin， PTX)或一系列激酶抑制劑處理后，我們采用ELISA法測定ARPE-19細(xì)胞上清液中IL-8、IL-6和MCP-1的質(zhì)量濃度水平，以及采用Western-Blot方法測定ARPE-19細(xì)胞內(nèi)絲裂原活化蛋白激酶信號傳導(dǎo)分子的活化情況和S1P3受體的表達(dá)水平。 結(jié)果: ARPE-19細(xì)胞表達(dá)所有已知的S1P1-5受體。此外，外源性的S1P促進(jìn)ARPE-19細(xì)胞分泌IL-8，且在一定范圍內(nèi)呈現(xiàn)劑量和時(shí)間依賴方式，但S1P并不能促進(jìn)IL-6和MCP-1的分泌。S1P誘導(dǎo)ARPE-19細(xì)胞分泌IL-8受PTX、ERK1/2和p38MAPK的調(diào)控。有趣的是，當(dāng)ARPE-19細(xì)胞被TNF-α刺激后，能上調(diào)ARPE-19細(xì)胞S1P3受體的表達(dá)，而且還能增加S1P促細(xì)胞分泌炎癥細(xì)胞因子IL-8和IL-6，但對MCP-1沒有影響。 結(jié)論:本研究表明，，S1P能顯著促進(jìn)ARPE-19細(xì)胞分泌炎癥細(xì)胞因子。ERk1/2、p38MAPK和Gi蛋白依賴通路是S1P促進(jìn)ARPE-19細(xì)胞分泌的重要信號成分。此外，這些結(jié)果還證明在眼部炎癥中，S1P對RPE細(xì)胞的刺激作用可能通過對白細(xì)胞功能的調(diào)控來發(fā)揮致炎作用。
[Abstract]:Aim: to investigate the effect of sphingosine 1 phosphate (S1P) on the secretion of inflammatory cytokines by ARPE-19 cells and the role of cell signal transduction pathway. Methods: the expression of S1P1-5 receptor in ARPE-19 cells was detected by RT-PCR and realtime PCR. ARPE-19 cells were treated with TNF- 偽, S1P, pertussis toxin (Pertussiutoxin, PTX) or a series of kinase inhibitors. The mass concentrations of IL-8,IL-6 and MCP-1 in the supernatant of ARPE-19 cells were determined by ELISA assay. The activation of mitogen-activated protein kinase signal transduction molecules and the expression of S1P3 receptor in ARPE-19 cells were measured by Western-Blot method. Results: ARPE-19 cells expressed all known S1P1-5 receptors. In addition, exogenous S1P promoted the secretion of IL-8, by ARPE-19 cells in a dose-and time-dependent manner, but S1P did not promote the secretion of IL-6 and MCP-1. S1P induced the secretion of IL-8 by PTX, by ARPE-19 cells. Regulation of ERK1/2 and p38MAPK. Interestingly, when ARPE-19 cells were stimulated by TNF- 偽, the expression of S1P3 receptor in ARPE-19 cells was up-regulated, and S1P stimulated the secretion of inflammatory cytokines IL-8 and IL-6, but had no effect on MCP-1. Conclusion: S1P can significantly promote the secretion of inflammatory cytokines in ARPE-19 cells. ERk1/2,p38MAPK and Gi protein-dependent pathway are important signal components of S1P in promoting the secretion of ARPE-19 cells. These results also suggest that S1P stimulates RPE cells through the regulation of leukocyte function in ocular inflammation.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R774.1

【共引文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前3條

1 李夢婷;鞘氨醇激酶1對結(jié)腸癌細(xì)胞血管生成擬態(tài)的影響及其機(jī)制[D];廣西醫(yī)科大學(xué);2013年

2 張智超;S1P通過S1PR1/3對THP-1源性巨噬細(xì)胞膽固醇流出的調(diào)控作用及機(jī)制研究[D];南華大學(xué);2013年

3 景小東;冠脈支架內(nèi)再狹窄與S1P的相關(guān)性研究[D];重慶醫(yī)科大學(xué);2013年

本文編號：2368970