黑骨藤乙醇提取物影响人类风湿关节炎滑膜成纤维细胞增殖及COX-2、PGE-2表达的研究  

Effects of Periploca forrestii ethanol extract( PFE) on proliferation,COX-2 expression,and PG2 expression of Rheumatoid arthritis synovial fibroblasts from patients

作　　者：梁江[1] 马武开[1] 刘正奇[1] 安阳[1] 姚血明[1] 唐志宇[1] 

机构地区：[1]贵阳中医学院,贵州贵阳550002

出　　处：《时珍国医国药》2015年第1期24-27,共4页Lishizhen Medicine and Materia Medica Research

基　　金：贵州省科学技术基金项目(黔科合J字[2013]2074号);2012年贵阳中医学院博士启动基金项目;国家自然科学基金(地区项目)(H2720);贵州省科技创新人才团队项目[黔科合人才团队(2013)4033]

摘　　要：目的观察黑骨藤乙醇提取物(Periploca forrestii ethanol extract,PFE)对人类风湿关节炎滑膜成纤维细胞(Rheumatoid arthritis synovial fibroblasts,RASF)的增殖以及环氧化酶-2(cyclooxygenase-2,COX-2)、前列腺素2(prostaglandin E2,PGE2)的表达影响。方法离体培养第4代的人类风湿关节炎滑膜成纤维细胞,同时设立空白对照组和试验组,两组细胞均先予白介素-1β(interleukin-1β,IL-1β)干预刺激,试验组予不同浓度的PFE(125,250,500μg/ml)干预细胞,24h后,采用MTT法检测细胞抑制率。用RT-RCR法测定COX-2的mRNA表达。免疫蛋白印记(WB法)检测COX-2的表达量,ELISA法检测48h末,细胞培养上清中PGE2的含量,结果干预48h后,PFE对细胞抑制率呈量-效果正相关性,各剂量组的PFE对PGE2、COX-2mRNA及其蛋白的表达量有明显抑制(P<0.05),结论 PFE能抑制IL-1β诱导的RASF增殖,在mRNA和蛋白两个层面均对RASF高表达的COX-2有明显抑制效果,同时对该细胞分泌的炎症效应分子PGE2也有抑制作用,这可能是黑骨藤治疗类风湿关节炎的潜在机制之一。Objective To investigate the effects of Periploca forrestii ethanol extract on proliferation, COX -2 expression, and PGE2 expression of Rheumatoid arthritis synovial fibroblasts from patients （RASF）. Methods RASF were separated from synovia and cultured in vitro, then divided into 5 groups , the blank control, positive control（ LL - 1β10ng/ml）, other 3 groups were first treated with LL- 1β（ lOng/ml）then deferent doses of PFE（ 125,250,500txg/ml） ,the level of RASF proliferation at 2 time point （24h,48h） were measured by MTT . After 48h,expression of COX- 2mRNA were measured by RT- PCR , expression of COX- 2 protein level were measured by Western Blot assay, expression of PGE2 in the culture supernatant were tested By ELISA. Results After 48h, compared with the positive control, different concentrations of HPE could significantly inhibit the proliferation of RASF with dose - effect relationship（ P 〈 0.05 ）, high and middle dose groups of PFE could significantly inhibit the expression of COX - 2 on mRNA and proteins level（ P 〈 0.05 ）. Conclusion The therapeutic mechanism of PFE for RA may include two aspects, first PFE could restrain proliferation of RASF ,second PFE may down regulate the expression of COX- 2 ,PGE2.

关 键 词：黑骨藤 类风湿滑膜成纤维细胞 环氧化酶-2 

分 类 号：R285.5[医药卫生—中药学]