岗松中抗前列腺素E2受体4拮抗剂筛选及其体内抗类风湿性关节炎作用  

Screening of prostaglandin E2 receptor 4 antagonist from active compounds of Baeckea frutescens L.and its anti-rheumatoid arthritis effect in vivo

作　　者：蔺小兵 汪豪[2] 张钧涵 林子豪 严明[1] 安晓飞 LIN Xiaobing;WANG Hao;ZHANG Junhan;LIN Zihao;YAN Ming;AN Xiaofei(Jiangsu Key Laboratory of Drug Screening,China Pharmaceutical University,Nanjing 210009;Department of Natural Medicinal Chemistry,China Pharmaceutical University,Nanjing 210009;Department of Endocrinology,Jiangsu Provincal Hospital of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]中国药科大学新药筛选中心,南京210009 [2]中国药科大学天然药物化学教研室,南京210009 [3]南京中医药大学附属医院内分泌科,南京210029

出　　处：《中国药科大学学报》2021年第1期92-99,共8页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81774248)。

摘　　要：筛选岗松中抗前列腺素E2受体4(EP4)的拮抗剂和探讨抗类风湿关节炎的作用。体外建立HEK293T-EP4细胞拮抗剂筛选模型,采用均相时间分辨荧光(HTRF)技术筛选岗松活性成分。SPF级别ICR小鼠,雄性,随机分为对照组、模型组、甲氨蝶呤组、岗松提取物BF-2(100、50和25 mg/kg)组,体内建立胶原诱导型类风湿性关节炎(CIA)小鼠模型,测量小鼠足趾肿胀体积,苏木精和伊红染色进行病理学检测。小鼠随机分为对照组、岗松提取物BF-2(100、50和25 mg/kg)组和阿司匹林组,采用醋酸诱导扭体试验观察扭体次数。本研究成功建立了EP4体外拮抗剂筛选模型,初筛结果发现岗松提取物BF-2,BF-20,BF-11和BF-12具有较强的EP4拮抗活性[(102.11±3.45)%,(90.31±3.59)%,(75.72±1.79)%和(76.84±1.64)%],其中,BF-2拮抗活性最强(IC50=0.99±0.08μg/mL)。BF-2可以明显抑制CIA小鼠的足趾肿胀,并缓解关节软骨基质降解和炎性细胞浸润。同时,与对照组比较,BF-2各剂量组小鼠的扭体次数均显著减少。筛选得到了一种EP4拮抗剂BF-2,并且具有潜在抗类风湿性关节炎活性。The objectives of the study were to screen prostaglandin E2 receptor 4 antagonist from active compounds of Baeckea frutescens L.and to explore its anti-rheumatoid arthritis effect.The HEK293T-EP4 cell antagonist screening model was established in vitro.Homogeneous time-resolved fluorescence(HTRF)technique was used to screen the active compounds of Baeckea frutescens L..SPF grade ICR male mice were randomly divided into control group,model group,methotrexate group,and Baeckea frutescens L.compound BF-2(100,50 and 25 mg/kg)groups.The collagen-induced arthritis(CIA)mouse model was established in vivo.The swelling volume of the toes of mice was measured,and the pathological examination was analyzed by staining with hematoxylin and eosin.SPF grade ICR mice,male,were randomly divided into control group,BF-2(100,50 and 25 mg/kg)group,and aspirin group.The acetic acid-induced body writhing test was observed.The EP4 in vitro antagonist screening model was successfully established.The preliminary screening results found that BF-2,BF-20,BF-11 and BF-12 had strong EP4 antagonistic activity[(102.11±3.45)%,(90.31±3.59)%,(75.72±1.79)%and(76.84±1.64)%],and BF-2 had the strongest antagonistic activity(IC50=0.99±0.08μg/mL).BF-2 could significantly inhibit the toe swelling of CIA mice,and relieve the degradation of articular cartilage matrix and inflammatory cell infiltration.At the same time,compared with the control group,the writhing times of the mice in each dose of BF-2 were significantly reduced.In this study,BF-2 of Baeckea frutescens L.was selected as an EP4 antagonist,which has potential anti-rheumatoid arthritis activity.

关 键 词：岗松 类风湿性关节炎 前列腺素E2受体4 炎症反应 

分 类 号：R965[医药卫生—药理学]