S1P对视网膜色素上皮细胞分泌炎症细胞因子的影响
发布时间：2018-12-08 20:43
【摘要】：目的:探讨1-磷酸鞘氨醇(S1P)对ARPE-19细胞分泌炎性细胞因子的影响以及细胞信号传导通路在其中的作用。 方法: ARPE-19细胞中S1P1-5受体的表达通过RT-PCR和realtime PCR进行检测。ARPE-19细胞经TNF-α、S1P、百日咳毒素(Pertussiutoxin， PTX)或一系列激酶抑制剂处理后，我们采用ELISA法测定ARPE-19细胞上清液中IL-8、IL-6和MCP-1的质量浓度水平，以及采用Western-Blot方法测定ARPE-19细胞内丝裂原活化蛋白激酶信号传导分子的活化情况和S1P3受体的表达水平。 结果: ARPE-19细胞表达所有已知的S1P1-5受体。此外，外源性的S1P促进ARPE-19细胞分泌IL-8，且在一定范围内呈现剂量和时间依赖方式，但S1P并不能促进IL-6和MCP-1的分泌。S1P诱导ARPE-19细胞分泌IL-8受PTX、ERK1/2和p38MAPK的调控。有趣的是，当ARPE-19细胞被TNF-α刺激后，能上调ARPE-19细胞S1P3受体的表达，而且还能增加S1P促细胞分泌炎症细胞因子IL-8和IL-6，但对MCP-1没有影响。 结论:本研究表明，，S1P能显著促进ARPE-19细胞分泌炎症细胞因子。ERk1/2、p38MAPK和Gi蛋白依赖通路是S1P促进ARPE-19细胞分泌的重要信号成分。此外，这些结果还证明在眼部炎症中，S1P对RPE细胞的刺激作用可能通过对白细胞功能的调控来发挥致炎作用。
[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- 伪, 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- 伪, 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.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R774.1
本文编号：2368970
[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- 伪, 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- 伪, 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.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R774.1
【共引文献】
相关硕士学位论文 前3条
1 李梦婷;鞘氨醇激酶1对结肠癌细胞血管生成拟态的影响及其机制[D];广西医科大学;2013年
2 张智超;S1P通过S1PR1/3对THP-1源性巨噬细胞胆固醇流出的调控作用及机制研究[D];南华大学;2013年
3 景小东;冠脉支架内再狭窄与S1P的相关性研究[D];重庆医科大学;2013年
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