摘要
目的通过Ⅱ型胶原诱导关节炎(CIA)小鼠模型探讨痛痹颗粒对类风湿关节炎的作用机制。方法将60只雄性小鼠随机分为6组,即正常组、模型组、高剂量组、中剂量组、低剂量组及雷公藤多苷片组,通过2次注射0.1 mL CⅡ乳剂进行免疫刺激,构建CIA模型,造模成功后分别按照50、25、12.5 g/kg浓度对痛痹颗粒高、中、低剂量组灌胃给药,并对模型组及正常组给予等体积生理盐水、雷公藤多苷片组予0.12 g/kg浓度的雷公藤多苷片混悬液,每日1次,持续22 d。采用螺旋测微器观察痛痹颗粒对CIA小鼠足肿胀的改善程度,MTT法检测脾细胞增殖及粘附能力,放射免疫法对滑膜细胞培养上清液中IL-1β、TNF-α含量进行检测,Western blot法检测滑膜组织中NLRP3炎性小体的表达。结果与正常组相比,模型组2次免疫后第13天就出现关节肿胀表现;与模型组比,痛痹颗粒高、中剂量组及雷公藤多苷片组均可明显减轻CIA模型小鼠的足肿胀程度、降低关节炎发生率与关节评分、抑制脾细胞增殖与粘附能力,并且抑制培养的滑膜细胞增殖以及降低滑膜细胞培养上清液中IL-1β、TNF-α含量,同时可抑制滑膜组织中NLRP3炎性小体的表达。结论痛痹颗粒高、中剂量组均可能通过抑制NLRP3炎性小体的表达,从而抑制滑膜细胞增殖,调节炎性因子生成,进而抑制类风湿关节炎的炎性反应,以达到减轻关节损伤的作用。
OBJECTIVE To explore the mechanisms of Tongbi Granules on rheumatoid arthritis(RA)in the typeⅡcollagen-induced arthritis(CIA)mice model.METHODS 60 male mice were randomly divided into 6 groups(n=10),including normal group,blank control group,high dose group,medium dose group,low dose group and tripterygium polyglycoside group.The CIA model was established by two injections of 0.1 mL CⅡemulsion.Then,the mice were administered with high,medium and low doses of Tongbi Granules of 50 g/kg,25 g/kg and 12.5 g/kg,respectively,after the model was established successfully.While,the model group and the normal group were given the same volume of saline,tripterygium glycoside group were given 0.12 g/kg tripterygium glycoside suspension,which once a day and sustained 22 days.The degree of foot swelling in CIA mice was observed by helical micromanometer.The proliferation and adhesion of spleen cells and T lymphocytes were detected by MTT assay.The effects of radiotherapy on the proliferation of synovial cells containing IL-1β,TNF-αlevels were detected.Western blot was used to detect the expression of NLRP3 inflammatory bodies in synovial tissues.RESULTS Compared with the normal group,the swelling of the joint appeared on the 13th day after the second immunization.Compared with the blank control group,the high dose group and the tripterygium polyglycoside group could significantly reduce the degree of foot swelling in the CIA model mice,and could significantly inhibit the spleen cell proliferation and its adhesion ability,while still inhibiting the culture Synovial cell proliferation and reduce the synovial cell culture supernatant contained in the IL-1β,TNF-αcontent.Also,it inhibited the expression of NLRP3 inflammatory body in synovial tissue.CONCLUSION The high and medium dose groups of Tongbi Granules may inhibit the expression of NLRP3 inflammatory body to reduce the proliferation of synovial cells and regulate the formation of inflammatory factors,thereby inhibiting the inflammatory response of RA and relieving joint injury.
作者
何晓瑾
王磊
孔甜
王瑞瑞
谢国倩
管玉洁
周格选
HE Xiao-jin;WANG Lei;KONG Tian;WANG Rui-rui;XIE Guo-qian;GUAN Yu-jie;ZHOU Ge-xuan(Jiangsu Hospital of Traditional Chinese Medicine, Affiliated to Nanjing University of Chinese Medicine, Nanjing, 210029, China;Liutan Community Health Service Center, Wuxi, 214000, China)
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2020年第2期229-234,共6页
Journal of Nanjing University of Traditional Chinese Medicine
基金
江苏省自然科学基金(BK20191415)
第三批中医临床优秀人才研修项目(苏中医科教〔2017〕18号)
江苏高校“青蓝工程”中青年学术带头人资助项目(苏教师〔2018〕1号)。