摘要
目的观察少阳膝痹方对大鼠膝骨关节炎的干预效应,初步从LXR-α/NF-κB信号通路探讨其可能的作用机制。方法采用改良hulth法建立大鼠双侧膝关节膝骨关节炎模型,随机分为假手术组、模型组、美洛昔康组、低、中、高剂量中药复方组,模型制备成功后灌胃给药,给药4周后观察软骨表面大体形态学,HE染色观察软骨病理变化,ELISA法检测血清中基质金属蛋白抑制剂-1(TIMP-1)、前列腺素2E(PGE2)、基质金属蛋白13(MMP13)指标,Western Blot检测大鼠膝关节TNF-α、调节肝X受体α(LXR-α)、核因子κB(NF-κB)P65、NF-κB P-P65蛋白的表达;建立膝关节软骨细胞培养体系,通过LXR-α激动剂及抑制剂干预,分为空白对照组、模型组、中药复方组、中药复方+抑制剂组、中药复方+激动剂组,检测PGE2、MMP13及TNF-α的表达情况。结果术后4周,大体形态学观察显示中药复方各组软骨表面较模型组更完整,HE染色显示各中药复方组关节软骨略有损伤,潮线略有杂乱,骨皮质略有损伤,较模型组有所改善。与假手术组比较,模型组MMP13浓度、PGE2、TNF-α蛋白表达、P-P65蛋白表达、P-P65/P65相对蛋白表达升高(P<0.05),NF-κB P65蛋白表达下降(P<0.05)。与模型组比较,美洛昔康组、各剂量中药复方组MMP13浓度、P-P65蛋白表达、P-P65/P65蛋白相对表达降低,TIMP-1浓度、TIMP-1/MMP13比值、LXR-α基因及蛋白表达升高(P<0.05);美洛昔康组、低、高剂量中药复方组PGE2、TNF-α蛋白降低,中剂量中药复方组TNF-α蛋白表达量亦降低(P<0.05)。与空白对照组比较,模型组MMP13、PGE2蛋白表达升高(P<0.05);与模型组比较,中药复方组及中药复方+激动剂组MMP13、PGE2蛋白表达量降低(P<0.05),TNF-α基因表达升高(P<0.05);与中药复方+抑制剂组比较,中药复方组、中药复方+激动剂组MMP13、PGE2表达量降低(P<0.05),TNF-α基因表达降低(P<0.05)。结论少阳膝痹方可降低血清中的MMP13、PGE
Objective To evaluate the intervention effect of Shaoyang Xibi Decoction(SXD)on rat knee osteoarthritis and to initiate an exploration into its probable mechanism of action through the LXR-α/NF-κB signaling pathway.Methods A rat bilateral knee osteoarthritis model was established by the modified Hulth technique and was subsequently segregated into six groups:sham operation group,model group,meloxicam group,and low,medium,and high dose Chinese medicine compound group.Following the successful preparation of the model,the drug was administered via gavage.After 4 weeks of treatment,the gross morphology of the cartilage surface was examined,while HE staining was employed to observe pathological changes in cartilage.ELISA was used to assay serum matrix metalloproteinase inhibitor-1(TIMP-1),prostaglandin E2(PGE2),and matrix metalloproteinase 13(MMP13)levels.Western Blot analysis was utilized to detect rat knee joint TNF-α,liver X receptorα(LXR-α),nuclear factorκB(NF-κB)P65,and NF-κB P-P65 proteins expressions.A knee joint cartilage cell culture system was established and intervened with LXR-αagonist and inhibitor.The cells were divided into blank controlgroup,model group,Chinese medicine compound group,Chinese medicine compound+agonist group and Chinese medicine compound+inhibitor group.The expression of PGE2 and TIMP-1 proteins were measured.Results Four weeks after surgery,gross morphology observations indicated that the cartilage surface integrity in the Chinese medicine compound groups was superior to that of the model group.HE staining revealed mild damage in joint cartilage,slightly disordered tide lines,and minor damage to the bone cortex within each Chinese medicine compound group,showing an improvement in comparison to the model group.Compared with the sham operation group,the model group exhibited increased MMP13 concentration,PGE2,TNF-αprotein expression,P-P65 protein expression,P-P65/P65 relative protein expression(P<0.05),and a decrease in NF-κB P65 protein expression(P<0.05).Compared with the mo
作者
张轩鸣
郭杨
潘娅岚
刘孟敏
涂鹏程
陆超然
马勇
ZHANG Xuan-ming;GUO Yang;PAN Ya-lan;LIU Meng-min;TU Pengcheng;LU Chao-ran;MA Yong(New Technology Laboratory of Bone Trauma Repair and Reconstruction,Nanjing University of Traditional Chinese Medicine,Nanjing,210023;Chronic Disease Nursing Intervention Laboratory,Nanjing University of Traditional Chinese Medicine,Nanjing,210023;College of Traditional Chinese Medicine,College of Integrated Traditional and Western Medicine,Nanjing University of Traditional Chinese Medicine,Nanjing,210023)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2023年第12期1469-1477,共9页
Chinese Journal of Integrated Traditional and Western Medicine
基金
江苏省自然科学基金面上项目(No.BK20211300)
江苏高校中西医临床医学品牌专业建设工程资助项目(二期)(No.2020PPZXL261)。
关键词
少阳膝痹方
膝骨关节炎
少阳主骨
肝X受体
核因子ΚB
中药复方
Shaoyang Xibi Decoction
knee osteoarthritis
Shaoyang governing bones
liver X receptor
nuclear factorκB
Chinese herbal compound
