摘要
目的观察和痹方剂对人类风湿关节炎(RA)成纤维样滑膜细胞(FLS)炎症因子分泌的影响,并探讨其机制。方法取FLS,并随机分为6组。激动剂组每孔加含10%PNU-282987药物血清的DMEM培养液,MTX组加含10%甲氨蝶呤(MTX)药物血清的DMEM培养液,低剂量组加含10%低剂量和痹方剂药物血清的DMEM培养液,中剂量组加含10%中剂量和痹方剂药物血清的DMEM培养液,高剂量组加含10%高剂量和痹方剂药物血清的DMEM培养液,模型组加10%DMEM血清的培养液。24 h后取细胞上清液及滑膜细胞。采用ELISA法检测细胞上清液IL-17、高迁移率组蛋白1(HMGB1),Western blotting法检测细胞α7烟碱型乙酰胆碱受体(α7nAChR)、核转录因子-κB亚基p65(NF-κB p65),RT-PCR法检测细胞α7nAchR mRNA。结果与模型组比较,激动剂组、MTX组、中剂量组IL-17、HMGB1表达降低(P均<0.05);与激动剂组比较,MTX组、低剂量组、中剂量组、高剂量组IL-17表达及高剂量组HMGB1表达升高(P均<0.05);与MTX组比较,中剂量组IL-17表达降低(P均<0.05)。与模型组比较,α7nAChR蛋白、mRNA表达升高,p65蛋白表达降低(P均<0.05);与激动剂组比较,低剂量组p65蛋白及中剂量组α7nAChR蛋白、p65蛋白表达升高,低剂量组、高剂量组α7nAChR蛋白及低剂量组、中剂量组、高剂量组α7nAChR mRNA表达降低(P均<0.05);与MTX组比较,低剂量组p65蛋白及中剂量组α7nAChR蛋白、NF-kBp65蛋白、α7nAChR mRNA表达升高(P均<0.05);与低剂量组比较,中剂量组α7nAChR蛋白、mRNA表达升高,中剂量组p65蛋白及高剂量组α7nAChR蛋白、p65蛋白表达降低(P均<0.05);与中剂量组比较,高剂量组α7nAChR、NF-kBp65蛋白表达降低(P均<0.05)。结论中等浓度和痹方剂能抑制RA FLS分泌炎症因子,进而起到治疗作用,机制可能为上调α7nAChR mRNA、蛋白表达,激活CAP,进而调节NF-kB信号通路。
Objective To observe the effect of Hebi recipe on the secretion of inflammatory factors in human rheumatoid arthritis(RA)fibroblast-like synoviocytes(FLSs)and to explore its mechanism.Methods FLSs were taken and randomly divided into 6 groups.In the agonist group,DMEM medium containing 10%PNU-282987 drug serum was added to each well;in the MTX group,DMEM medium containing 10%methotrexate(MTX)drug serum was added;in the lowdose group,DMEM medium containing 10%low-dose Hebi recipe drug serum was added;in the medium-dose group,DMEM medium containing 10%medium-dose Hebi recipe drug serum was added;in the high-dose group,DMEM medium containing 10%high-dose Hebi recipe drug serum was added;and in the model group,10%DMEM serum was added.After 24 h,the cell supernatant and synovial cells were taken.The interleukin(IL)-17 and high mobility group box 1(HMGB1)in the supernatant were detected by enzyme linked immunosorbent assay(ELISA),α7 nicotinic acetylcholine receptor(α7 nAChR)and nuclear transcription factor-κB subunit p65(NF-κB p65)were detected by Western blotting,and theα7 nAchR mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Compared with the model group,the expression of IL-17 and HMGB1 in the agonist group,the MTX group and the medium-dose group decreased(both P<0.05).Compared with the agonist group,the expression of IL-17 in the MTX group,the lowdose group,the medium-dose group,the high-dose group,and the expression of HMGB1 in the high-dose group increased(both P<0.05).Compared with the MTX group,the IL-17 expression decreased in the medium-dose group(P<0.05).Compared with the model group,the protein and mRNA expression ofα7 nAChR in both the agonist group and the mediumdose group increased,and the expression of p65 protein decreased(all P<0.05).Compared with the agonist group,the expression of p65 protein in the low-dose group and the expression ofα7 nAChR protein and p65 protein in the medium-dose group increased,the expression ofα7 nAChR protein in the low-dose grou
作者
姜萍
邢洁
纪淳望
刘英
马洪美
姜月华
JIANG Ping;XING Jie;JI Chunwang;LIU Ying;MA Hongme;JIANG Yuehua(The First Clinical Medical College of Shandong University of Traditional ChineseMedicine,Jinan 250014,China)
出处
《山东医药》
CAS
2019年第3期39-42,共4页
Shandong Medical Journal
基金
山东省科技发展计划项目(2014GSF119017)
国家中医药管理局第四批全国中医(临床
基础)优秀人才研修项目(国中医药教发[2017]24)
关键词
和痹方剂
类风湿关节炎
成纤维样滑膜细胞
炎症因子
Hebi recipe
rheumatoid arthritis
fibroblast-like synoviocytes
inflammatory factor
