Essential Roles of Sphingosine 1-Phosphate Receptor Types 1 and 3 in Human Hepatic Stellate Cells Motility and Activation
Essential Roles of Sphingosine 1-Phosphate Receptor Types 1 and 3 in Human Hepatic Stellate Cells Motility and Activation
复制标题
1 型和 3 型鞘氨醇 1-磷酸受体在人肝星状细胞运动和激活中的重要作用
DOI：
10.1002/jcp.22572
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发表时间：
2011-09-01
影响因子：
5.6
通讯作者：
Li, Liying
中科院分区：
文献类型：
--
作者：
Liu, Xihong;Yue, Shi;Li, Liying
1-磷酸鞘氨醇（S1 P）及其受体（S1 PRs）的生物学作用已被广泛研究。然而，目前S1 P/S1 PRs轴在肝纤维化中的病理生理作用尚不清楚。本研究以人肝星状细胞系LX-2细胞为研究对象，探讨了S1 P/S1 PRs轴的功能。RT-PCR、Westernblot和免疫细胞化学分析表明，LX-2细胞中存在S1 PR 1 -3型。S1 P对LX-2细胞产生了强有力的迁移作用，如在Boyden室中测定的，并刺激LX-2细胞的纤维化活性，如通过平滑肌α-肌动蛋白、前胶原α 1（I）和α 1（III）以及总羟脯氨酸含量的表达增加所证明的。此外，S1 P的作用被S1 PR 1激动剂SEW 2871模拟，并且被W146（S1 PR 1拮抗剂）和/或沉默S1 PR 1消除，三种表达用小干扰RNA，表明S1 PR 1和3的主要作用。然而，S1 PR 2拮抗剂JTE-013和沉默S1 PR 2表达的研究表明，S1 PR 2负调控S1 P诱导的细胞迁移。有趣的是，外源性添加S1 P诱导鞘氨醇激酶-1和合成的额外的S1 P的显着上调，和S1 PR 1，3的表达，但不是S1 PR 2。总之，我们的数据已经确定了由S1 P/S1 PR 1，3轴调节的另外的功能，涉及HSC的迁移和纤维化激活。这些结果表明，选择性调节S1 PR活性可能代表一种新的抗纤维化策略。J.细胞。226：2370-2377，2011。(C)2010 Wiley-Liss，Inc.
The biological roles of sphingosine 1-phosphate (S1P) and S1P receptors (S1PRs) have been broadly investigated. However, at present pathophysiological roles of S1P/S1PRs axis in liver fibrosis are not well defined. Here, we investigated the functions of S1P/S1PRs axis in human hepatic stellate cells (HSC) line, LX-2 cells. We found that S1PR types 1, 2 and 3 (S1PR1-3) are clearly detected in LX-2 cells, as determined by RT-PCR, Western blot and immunocytochemistry analysis. S1P exerted a powerful migratory action on LX-2 cells, as determined in Boyden chambers, and stimulated fibrogenic activity of LX-2 cells, as demonstrated by increase of expression of smooth muscle a-actin, procollagen alpha 1(I) and alpha 1(III) and total hydroxyproline content. Moreover, the effects of S1P were mimicked by S1PR1 agonist SEW2871, and abrogated by W146 (S1PR1 antagonist) and/or silencing S1PR1, three expression with small interfering RNA, suggesting the main roles of S1PR1 and 3. However, studies with S1PR2 antagonist JTE-013 and silencing S1PR2 expression indicated that S1PR2 negatively regulated S1P-induced cell migration. Interestingly, exogenously added S1P induced significant up-regulation of sphingosine kinase-1 and the synthesis of additional S1P, and expression of S1PR1,3, but not S1PR2. In conclusion, our data have identified an additional function regulated by S1P/S1PR1,3 axis involving migration and fibrogenic activation of HSCs. These results suggest that selective modulation of S1PR activity may represent a new antifibrotic strategy. J. Cell. Physiol. 226: 2370-2377, 2011. (C) 2010 Wiley-Liss, Inc.