PGE_(2)通过EP4/PKA信号通路调控滑膜细胞OAT1的表达及CP-25的作用  

PGE_(2) regulates expression of OAT1 in synoviocytes via EP4/PKA signaling pathway and effect of CP-25

作　　者：肖康俊 高锦张 王勇 王斌[1] 魏伟[1] 王春[1] XIAO Kang-jun;GAO Jin-zhang;WANG Yong;WANG Bin;WEI Wei;WANG Chun(Institute of Clinical Pharmacology,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immuno Drugs,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,安徽省抗炎免疫药物协同创新中心,安徽合肥230032

出　　处：《中国药理学通报》2024年第9期1658-1664,共7页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金资助项目(No 2008085QH402)。

摘　　要：目的明确前列腺素E2(prostaglandin E2,PGE_(2))对滑膜细胞有机阴离子转运体1(organic anion transporter 1,OAT1)膜表达的调控机制及芍药苷-6'-O-苯磺酸酯(CP-25)的作用。方法免疫荧光法检测不同浓度CP-25对PGE_(2)处理后滑膜细胞OAT1和前列腺素E受体4(prostaglandin E receptor 4,EP4)表达的影响;使用EP4激动剂(TCS2510)与拮抗剂(GW627368X),探究EP4在OAT1调节中的作用;使用CP-25和蛋白激酶A(protein kinase A,PKA)抑制剂H-89,探究CP-25和PKA对滑膜细胞OAT1表达的影响。结果PGE_(2)在0~10 min内明显下调EP4与OAT1的膜表达,20~60 min后明显上调(P<0.05);CP-25明显上调PGE_(2)处理后细胞膜OAT1和EP4的表达(P<0.05);EP4激动剂TCS2510明显上调细胞膜OAT1的表达(P<0.01);CP-25上调PGE_(2)处理的细胞中OAT1的表达,GW627368X和H-89均能下调PGE_(2)和CP-25处理的滑膜细胞中OAT1的表达(P<0.01)。结论PGE_(2)介导的EP4/PKA信号通路可以调控OAT1在滑膜细胞膜上的表达,CP-25可以通过活化EP4/PKA信号通路明显上调滑膜细胞中OAT1的膜表达。Aim To investigate the regulatory mechanism of prostaglandin E2(PGE_(2))on the membrane expression of organic anion transporter 1(OAT1)in synoviocytes and the effect of paeoniflorin-6'-O-benzene sulfonate(CP-25).Methods Immunofluorescence was used to detect the effects of CP-25 on the expression of OAT1 and prostaglandin E receptor 4(EP4)in PGE_(2) stimulated synoviocytes.EP4 agonists(TCS2510)and antagonists(GW627368X)were used to explore the role of EP4 in regulation of OAT1.CP-25 and protein kinase A(PKA)inhibitor H-89 were used to investigate the effects of CP-25 and PKA on OAT1 expression in synoviocytes.Results Membrane expression of EP4 and OAT1 was down-regulated within 0~10 min,and thereby up-regulated between 20~60 min in the presence of PGE_(2) simulation(P<0.05).CP-25 significantly up-regulated the expressions of OAT1 and EP4 in PGE_(2) stimulated synoviocytes(P<0.05).TCS2510 significantly up-regulated the expression of OAT1 in cell membrane of synoviocytes(P<0.01).CP-25 up-regulated the expression of OAT1 in PGE_(2)-treated cells,while the effect of CP-25 on expression of OAT1 was significantly inhibited after the together use of GW627368X and H-89.(P<0.01).Conclusions PGE_(2)-mediated EP4/PKA signaling pathway regulates the expression of OAT1 on membranes of synoviocytes.CP-25 can significantly increase the membrane expression of OAT1 in synoviocytes by activating the EP4/PKA signaling pathway.

关 键 词：芍药苷-6'-O-苯磺酸酯 滑膜细胞 有机阴离子转运蛋白1 前列腺素E2 前列腺素E受体4 蛋白激酶A 

分 类 号：R-332[医药卫生] R284.1R322.72R345.57R593.22R977.6