CP-25对大鼠胶原性关节炎OAT1的调控作用及机制  

The regulatory role and mechanism of CP-25 on OAT1in rats with collagen-induced arthritis

作　　者：段菲 高锦张 王勇 王斌[1] 魏伟[1] 王春[1] DUAN Fei;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12期2295-2302,共8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 明确胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠滑膜有机阴离子转运蛋白1(organic anion transporter^(-1),OAT1)的表达变化及芍药苷-6'-O-苯磺酸酯(paeoniflorin-6'-O-benzenesulfonate,CP-25)的作用,并初步探讨其可能的调控机制。方法 建立大鼠CIA模型,给予CP-25、帕罗西汀、前列腺素E受体4 (prostaglandin E receptor 4,EP4)拮抗剂或CP-25+EP4拮抗剂联用治疗,甲氨蝶呤为阳性对照药。测量足爪容积,进行全身及关节炎指数评分,计算关节肿胀数,进行足爪X线摄片、踝关节病理检查及评分。ELISA测定血清前列腺素E_(2) (prostaglandin E_(2),PGE_(2))水平;Western blot测定CP-25用药后大鼠滑膜组织膜蛋白及总蛋白表达水平。结果 CP-25可减轻CIA大鼠足爪肿胀情况,降低CIA大鼠全身评分、关节炎指数、足爪容积和肿胀关节数,能明显缓解CIA大鼠组织病理学改变及骨侵蚀程度。此外,CP-25能降低CIA大鼠血清PGE_(2)浓度。CIA大鼠滑膜组织OAT1和EP4膜表达均下调,CP-25或帕罗西汀处理增加了OAT1和EP4的表达水平,而EP4拮抗剂可抑制CP-25对OAT1的影响。此外,CIA模型大鼠滑膜组织p-GRK2表达上调;CP-25或帕罗西汀干预后p-GRK2表达均降低;PKA的表达趋势与OAT1相似。结论 OAT1在CIA大鼠滑膜组织中膜表达降低,CP-25治疗可上调OAT1的表达。PGE_(2)-EP4-PKA信号通路可能参与了CP-25对OAT1的调控。Aim The present study was conducted to investigate the expression of organic anion transporter 1(OAT1)in synovial tissue of collagen-induced arthritis(CIA)rats and the effect of paeoniflorin-6′-O-benzene sulfonate(CP-25),and meanwhile reveal its potential regulatory mechanism.Methods The rat CIA model was established and treated with CP-25(50 mg·kg^(-1)·d^(-1)),paroxetine(15 mg·kg^(-1)·d^(-1)),prostaglandin E receptor 4(EP4)antagonist(3 mg·kg^(-1)·d^(-1)),or a combination of CP-25(50 mg·kg^(-1)·d^(-1))plus EP4 antagonist(3 mg·kg^(-1)·d^(-1)),while methotrexate(0.5 mg·kg^(-1)·3d^(-1))was used as the positive control drug.Clinical manifestation of CIA rats,including arthritis index,numbers of swollen joints,paw volume of non-injected hind and clinical scores was measured.Moreover,X-ray imaging of paw,pathological examination of the ankle joint and quantitative assessment were conducted.The concentration of serum prostaglandin E_(2)(PGE_(2))was quantified using the ELISA.Western blot analysis was employed to assess the levels of phospho-G protein-coupled receptor kinase 2(p-GRK2),protein kinase A(PKA),EP4 and OAT1 membrane proteins of rat synovial tissues following CP-25 treatment.Results The administration of CP-25 resulted in a reduction in foot swelling,global score,arthritis index,paw volume of non-injected,and numbers of swollen joints in CIA rats.Pathological examination and X-ray analysis revealed that CP-25 significantly ameliorated the pathological changes and bone erosion degree of CIA rats.CP-25 significantly lowered PGE_(2) concentration in the serum of CIA rats.In the CIA model group,membrane expression of OAT1 and EP4 in synovial tissues of rats was significantly downregulated.Treatment with CP-25 or paroxetine resulted in a significant upregulation of OAT1 and EP4 expression and the effect of CP-25 on OAT1 was blocked when EP4 antagonist was used together.Furthermore,the expression of p-GRK2 in synovial tissue from CIA rats was significantly upregulated,and however,CP-25 or paroxetine in

关 键 词：类风湿性关节炎 芍药苷-6'-O-苯磺酸酯 有机阴离子转运蛋白1 前列腺素E_(2) 前列腺素E受体4 蛋白激酶A G蛋白偶联受体2 

分 类 号：R-332[医药卫生] R284.1R329.11R345.57R593.22