基于LXRs/NF-B通路探讨壮骨健膝方对兔膝骨关节炎滑膜组织炎症的影响  

Effect of Zhuanggu Jianxi Decoction on Synovitis in Knee Osteoarthritis Rabbits via LXRs/NF-κB Pathway

作　　者：陈鹏[1,2] 郭洁梅[1,3] 肖艳 王建辉[1] 苏友新 CHEN Peng;GUO Jiemei;XIAO Yan;WANG Jianhui;SU Youxin(Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Key Laboratory of Orthopedics&Traumatology and Rehabilitation of Traditional Chinese Medicine,Ministry of Education,Fuzhou,Fujian 350122,China;Fujian Health College,Fuzhou,Fujian 350101,China)

机构地区：[1]福建中医药大学,福建福州350122 [2]中医骨伤及运动康复教育部重点实验室,福建福州350122 [3]福建卫生职业技术学院,福建福州350101

出　　处：《康复学报》2022年第1期48-55,共8页Rehabilitation Medicine

基　　金：国家自然科学基金面上项目(81774347);福建省自然科学基金项目(2019J01091)。

摘　　要：目目的的:从兔滑膜组织的炎症调控及LXRs/NF-κB通路关键节点蛋白的表达情况探讨壮骨健膝方对兔膝骨关节炎(KOA)滑膜组织炎症的干预效应及机制。方法:制备兔空白血清及含药血清以备滑膜组织干预使用。将22只兔按照随机数字表法分为造模组16只和空白组6只,造模组予右后肢膝关节腔注射木瓜蛋白酶制备KOA模型,模型鉴定成功后,无菌操作下切取兔右后肢膝关节内滑膜组织,将其剪碎成大小约1~2 mm3小块,收集60小块,随机分为4组,即模型组、壮骨健膝方组、LXRα抑制剂组和N-CoR抑制剂组,每组15个组织块,每孔3块,每组5孔,置于6孔板中培养,分别给予10%空白血清、10%含药血清、10%含药血清+5CPPSS-50、10%含药血清+ML-792干预;空白组滑膜同法制备,给予10%空白血清培养,5孔。7 d后收集滑膜组织,各组取2孔滑膜组织块行HE染色,剩余滑膜组织行Western blot和ELISA检测相关指标。结果果:(1)在形态学方面,与空白组比较,模型组HE染色可见滑膜内衬层细胞增生且排列紊乱明显、大量炎性细胞浸润及纤维组织增生等改变,形态学评分显著升高(P<0.05);与模型组比较,壮骨健膝方组可见滑膜内衬层细胞轻度增生排列紊乱、稀疏散在炎性细胞浸润及少量纤维组织增生等改变,形态学评分显著降低(P<0.05);分别与LXRα抑制剂组及N-CoR抑制剂组比较,壮骨健膝方组病理改变形态学评分显著降低(P<0.05)。(2)在炎症相关指标方面,干预后与空白组比较,模型组IL-1β、TNF-α、MMP-3、MMP-13含量显著升高(P<0.05);与模型组比较,LXRα抑制剂组、N-CoR抑制剂组、壮骨健膝方组上述指标含量均显著降低(P<0.05);分别与LXRα抑制剂组及N-CoR抑制剂组比较,壮骨健膝方组上述指标含量均显著降低(P<0.05);壮骨健膝方组干预前后比较,IL-1β、TNF-α含量显著降低(P<0.05)。(3)在通路相关蛋白表达方面,与空白组比较,模型组LXRα、N-CoR蛋�Objective:To study the intervention effect and mechanism of Zhuanggu Jianxi decoction on synovitis in knee osteoarthritis(KOA)rabbits via the inflammatory regulation of synovial tissue and the expression of key node proteins in LXRs/NF-κB pathway.Methods:Blank serum and drug-containing serum of rabbits were prepared for the intervention of synovial tissue.Twentytwo rabbits were randomly divided into two groups,with 16 in the model group and 6 in the blank group.The KOA model was injected with papain into the knee joint of the right hind limb.After the model was identified successfully,the synovial tissue of the right hind limb knee was cut into small pieces with a size about 1-2 mm3 under aseptic operation,60 pieces were collected and randomly divided into model group,Zhuanggu Jianxi decoction group,LXRαinhibitor group and N-CoR inhibitor group with 15 pieces each,the synovial tissue pieces in each group were cultured in 6 well plates with every 3 pieces in each well,the four groups were intervened with 10%blank serum,10%drug-containing serum,10%drug-containing serum+5 CPPSS-50 and 10%drug-containing serum+ML-792 respectively;the synovium of the blank group was prepared by the same method and cultured with 10%blank serum for 5 wells.After 7 days,synovial tissue was collected.Two wells of synovial tissue were taken from each group for HE staining,and the remaining synovial tissue was detected by Western blot and ELISA.Results:1)Morphology:compared with the blank group,HE staining in the model group showed the proliferation and disorder of synovial lining cells,a large number of inflammatory cell infiltration and fibrous tissue hyperplasia,and the morphological score was significantly increased(P<0.05).Compared with the model group,mild hyperplasia and disorder of synovial lining cells,sparse scattered inflammatory cell infiltration and a small amount of fibrous tissue hyperplasia were observed in the Zhuanggu Jianxi decoction group,and the morphological score was significantly decreased(P<0.05).Compared with the

关 键 词：膝骨关节炎 滑膜组织炎症 壮骨健膝方 兔膝关节滑膜 LXRs/NF-κB通路 

分 类 号：R285.5[医药卫生—中药学]