AMP010014A09 in Sus Scrofa Encodes an Analog of G Protein-Coupled Receptor 109A, Which Mediates the Anti-Inflammatory Effects of Beta-Hydroxybutyric Acid
AMP010014A09 in Sus Scrofa Encodes an Analog of G Protein-Coupled Receptor 109A, Which Mediates the Anti-Inflammatory Effects of Beta-Hydroxybutyric Acid
复制标题
野猪中的 AMP010014A09 编码 G 蛋白偶联受体 109A 的类似物，介导 β-羟基丁酸的抗炎作用
DOI：
10.1159/000479206
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Liu, Juxiong
中科院分区：
文献类型：
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作者：
Chen, Guangxin;Fu, Shoupeng;Liu, Juxiong
背景：羟基羧酸受体2 （HCA2，也称为GPR109A）属于G蛋白偶联受体（GPCR）家族，存在于人类、大鼠、小鼠、仓鼠和豚鼠中，但在其他物种中几乎没有发现该蛋白的报道。在本研究中，我们推测AMP010014A09 （AMP+）是猪GPR109A的同源物。方法：为了验证这一假设，设计了以下实验：将猪外周血分离的单核细胞在加或不加β-羟基丁酸（BHBA）预处理后用LPS处理，并评估促炎细胞因子和炎症蛋白的水平。用BHBA处理或不处理，在AMP+基因沉默或稳定转染后，检测福斯可林诱导的猪睾丸（ST）和IPEC-J2细胞cAMP水平。结果：猪的AMP+与人的HM74A和小鼠的PUMA-G具有高度的同源性。BHBA抑制lps诱导的猪单核细胞促炎因子TNF-α、IL-6、IL-1β和炎性蛋白COX-2的分泌。BHBA抑制forskolin诱导的ST细胞cAMP水平升高，但用pGPU6-GFP-Neo-AMP+-su -392质粒转染细胞后，用shRNA沉默AMP+基因后，无法抑制cAMP的积累。用慢病毒载体转染AMP+基因后，BHBA对IPEC-J2细胞cAMP水平无影响，但显著抑制Forskolin处理诱导的cAMP水平升高。结论：我们的研究结果表明，AMP+在猪单核细胞中编码G蛋白偶联受体，抑制cAMP水平并介导抗炎作用。
Background: Hydroxy-carboxylic acid receptor 2 (HCA2, also called GPR109A) belongs to the G protein-coupled receptor (GPCR) family and is found in humans, rats, mice, hamsters and guinea pigs, but there are almost no reports of this protein in other species. In this investigation, we speculated that AMP010014A09 (AMP+) is a homologue of GPR109A in swine. Methods: To test this hypothesis, the following experiments were designed: monocytes isolated from the peripheral blood of swine were treated with LPS after pretreating with or without β-hydroxybutyric acid (BHBA), and the levels of pro-inflammatory cytokines and inflammatory proteins were assessed. cAMP levels induced by Forskolin in swine testicular (ST) and IPEC-J2 cells were detected with or without BHBA treatment and following silencing or stable transfection of the AMP+ gene. Results: AMP+ in swine exhibited a high level of homology with HM74A in humans and PUMA-G in mice. BHBA inhibited the LPS-induced secretion of the pro-inflammatory cytokines TNF-α, IL-6 and IL-1β and the inflammatory protein COX-2 in monocytes of swine. BHBA suppressed the Forskolin-induced cAMP level increase in ST cells, but failed to inhibit the accumulation of cAMP after the AMP+ gene was silenced with shRNA by transfecting cells with the pGPU6-GFP-Neo-AMP+-sus-392 plasmid. BHBA had no effect on cAMP levels in IPEC-J2 cells, but significantly inhibited the increase in cAMP induced by Forskolin treatment following transfection of the AMP+ gene into IPEC-J2 cells by a lentivirus vector. Conclusion: Our results indicated that AMP+ encodes a G protein-coupled receptor in Sus scrofa that inhibits cAMP levels and mediates anti-inflammatory effects in swine monocytes.