GPR35 mediates lodoxamide-induced migration inhibitory response but not CXCL17-induced migration stimulatory response in THP-1 cells; is GPR35 a receptor for CXCL17?
GPR35 mediates lodoxamide-induced migration inhibitory response but not CXCL17-induced migration stimulatory response in THP-1 cells; is GPR35 a receptor for CXCL17?
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DOI：
10.1111/bph.14082
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发表时间：
2018-01-01
影响因子：
7.3
通讯作者：
Im, Dong-Soon
中科院分区：
文献类型：
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作者：
Park, Soo-Jin;Lee, Seung-Jin;Im, Dong-Soon
背景和目的由于没有发现GPR 35的内源性配体，GPR 35一直被认为是一种孤儿GPCR。CXCL 17（一种趋化因子）已被报道是GPR 35的内源性配体，甚至有人建议将其称为CXCR 8。然而，目前还没有证据表明CXCL 17与GPR 35确实相互作用。实验方法我们采用两种检测系统来探讨CXCL 17与GPR 35之间的关系。使用GPR 35过表达HEK 293细胞中的AP-TGF-脱落测定作为功能获得测定。在内源性GPR 35表达THP-1 cells.Key ResultsIn the AP-TGF-alpha shedding assay，lodoxamide，一种众所周知的合成GPR 35激动剂，被证实是最有效的激动剂，在其他报道的激动剂中。然而，无论是人还是小鼠CXCL 17对GPR 35都没有影响。与先前的发现一致，发现G蛋白G（i/o）和G（12/13）与GPR 35偶联。此外，CID 2745687（一种选择性GPR 35拮抗剂）可浓度依赖性抑制洛度沙胺诱导的GPR 35激活。在内源性表达GPR 35的THP-1细胞中，洛度沙胺浓度依赖性地抑制迁移，并且这种抑制作用被CID 2745687处理或GPR 35 siRNA转染阻断。然而，即使CXCL 17刺激THP-1细胞的迁移，这是与以前的报告一致，CXCL 17的这种刺激作用没有被CID 2745687或GPR 35 siRNA阻断。结论和启示目前的研究结果表明，GPR 35作为一个迁移抑制受体的功能，但CXCL 17刺激的THP-1细胞的迁移是不依赖于GPR 35。
Background and PurposeGPR35 has long been considered an orphan GPCR, because no endogenous ligand of GPR35 has been discovered. CXCL17 (a chemokine) has been reported to be an endogenous ligand of GPR35, and it has even been suggested that it be called CXCR8. However, at present there is no supporting evidence that CXCL17 does interact with GPR35.Experimental ApproachWe applied two assay systems to explore the relationship between CXCL17 and GPR35. An AP-TGF- shedding assay in GPR35 over-expressing HEK293 cells was used as a gain-of-function assay. GPR35 knock-down by siRNA transfection was performed in endogenously GPR35-expressing THP-1 cells.Key ResultsIn the AP-TGF-alpha shedding assay, lodoxamide, a well-known synthetic GPR35 agonist, was confirmed to be the most potent agonist among other reported agonists. However, neither human nor mouse CXCL17 had an effect on GPR35. Consistent with previous findings, G proteins G(i/o) and G(12/13) were found to couple with GPR35. Furthermore, lodoxamide-induced activation of GPR35 was concentration-dependently inhibited by CID2745687 (a selective GPR35 antagonist). In endogenously GPR35-expressing THP-1 cells, lodoxamide concentration-dependently inhibited migration and this inhibitory effect was blocked by CID2745687 treatment or GPR35 siRNA transfection. However, even though CXCL17 stimulated the migration of THP-1 cells, which is consistent with a previous report, this stimulatory effect of CXCL17 was not blocked by CID2745687 or GPR35 siRNA.Conclusions and ImplicationsThe present findings suggest that GPR35 functions as a migration inhibitory receptor, but CXCL17-stimulated migration of THP-1 cells is not dependent on GPR35.