摘要
目的 探讨热补针法治疗类风湿关节炎(RA)寒证的抗炎机制。方法 采用卵蛋白干粉、弗氏完全佐剂联合低温冷冻法制备RA寒证家兔模型。按随机数字表法将50只家兔(雌雄各半)随机分为正常组、模型组、瑞沙托维(TAK-242)组、热补针法组及脂多糖(LPS)联合热补针法组,每组10只。TAK-242组腹腔注射Toll样受体4(TLR4)抑制剂TAK-242(0.75 mg/kg),热补针法组于“足三里”行热补针法针刺,LPS联合热补针法组行热补针法针刺合腹腔注射TLR4激动剂LPS(0.25 mg/kg)。连续治疗14 d后,比较各组家兔膝关节周径、皮肤温度;HE染色法观察家兔膝关节滑膜组织形态学;免疫组化法检测各组家兔膝关节滑膜组织中的肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)表达水平;Western blotting法检测各组家兔膝关节滑膜组织TLR4、髓样分化因子88(MyD88)、丝裂原活化蛋白激酶1/2(ERK1/2)的蛋白表达水平;实时荧光PCR检测各组家兔哺乳动物雷帕霉素靶蛋白(mTOR)、核糖体蛋白S6激酶(p70S6K)mRNA的表达。结果 与模型组比较,各干预组家兔膝关节周径缩小、皮肤温度升高(P<0.01);HE染色法示,TAK-242组及热补针法组家兔膝关节关节囊内侧黏膜基本平整,血管翳增殖减轻,存在少量炎性细胞浸润,而LPS联合热补针法组关节囊内侧黏膜粗糙,血管翳增殖,炎性细胞浸润聚集明显;滑膜组织中TNF-α、IL-1β、IL-6平均光密度均降低(P<0.01);TLR4、MyD88、ERK1/2蛋白表达水平及mTOR、p70S6K mRNA表达水平均降低(P<0.01)。与LPS联合热补针法组比较,热补针法组及TAK-242组家兔膝关节周径缩小、皮肤温度升高(P<0.01);滑膜组织中TNF-α、IL-1β、IL-6平均光密度均降低(P<0.01);TLR4、MyD88、ERK1/2蛋白表达水平及mTOR、p70S6K mRNA表达水平均降低(P<0.01)。结论 热补针法可以有效改善RA寒证家兔模型膝关节肿胀,缓解炎症反应,其机制可能与抑制滑膜组织TLR4
Objective We aimed to investigate the anti-inflammatory mechanism of heat-reinforcing needling on rheumatoid arthritis(RA) with cold syndrome.Methods The RA with cold syndrome model was established by injecting ovalbumin dry powder and Freund’s complete adjuvant combined with cold freezing.Fifty rabbits(50% male and 50% female) were randomly divided into the normal group, the model group, the resatorvid(TAK-242) group, the heat-reinforcing needling group, and the lipopolysaccharide(LPS) combined with heat-reinforcing needling group, 10 rabbits in each group.Rabbits in the TAK-242 group were intraperitoneally injected with the TLR4 inhibitor TAK-242(0.75 mg/kg);rabbits in the heat-reinforcing needling group were treated with “Zusanli”(ST 36) with heat-reinforcing needling;and rabbits in the LPS combined with heat-reinforcing needling group were injected intraperitoneally with the TLR4 agonist LPS(0.25 mg/kg) on the basis of heat-reinforcing needling. Each group received continuous treatment for 14 days. The circumference and skin temperature of the knee joint of the rabbits were recorded;the histomorphology of the synovial membrane of the rabbit knee joints was observed by HE staining;the expression levels of TNF-α, IL-1β, and IL-6 in synovial tissues were detected by immunohistochemistry;the protein expression levels of TLR4, MyD88, and ERK1/2 in synovial tissues were detected by Western blotting;and the mRNA levels of mTOR and p70 S6 K were detected by real-time PCR.Results The circumference of the knee joint was lower and the skin temperature of the knee joint was higher(P<0.01) in all intervention groups compared with the model group. The HE staining of the TAK-242 group and the heat-reinforcing needling group showed that the inner mucosa of the joint capsule was basically flat, the proliferation of pannus was reduced, and there was a small amount of inflammatory cell infiltration, while the LPS combined with the heat-reinforcing needling group showed that the inner mucosa of the joint capsule was roug
作者
苏成红
杜小正
方晓丽
刘强
刘莉梅
袁博
井维尧
张小娜
宋亚文
SU Chenghong;DU Xiaozheng;FANG Xiaoli;LIU Qiang;LIU Limei;YUAN Bo;JING Weiyao;ZHANG Xiaona;SONG Yawen(College of Acupuncture-Moxibustion and Tuina,Gansu University of Chinese Medicine,Lanzhou 730000,China;Department of Zheng-shi Acupuncture,Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2022年第7期745-754,共10页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金项目(No.82060891)
甘肃省自然科学基金项目(No.21JR7RA568)
国家中医药管理局甘肃郑氏针法学术流派传承工作室项目(No.2305135901)。
