摘要
目的:探讨山药多糖关节腔注射对兔膝关节骨性关节炎模型软骨退变的保护机制及抑制炎症因子表达水平的影响,为山药多糖的开发及研究提供相关证据,同时为进一步研究奠定基础。方法:将55只新西兰白兔,用随机数字表法分为5组,每组11只,采用改良Hulth法制备膝关节骨性关节炎兔模型,分别为模型组、山药多糖低、中、高剂量(0.7,1.43,2.15 mg·kg^(-1))组、玻璃酸钠(1.00 mg·kg^(-1))组。造模成功1周后模型组不干预治疗,各组使用相应药物干预5周,1次/周。5周后通过肉眼观察关节大体标本,光镜下观察关节软骨组织苏木素-伊红(HE)染色和甲苯胺蓝(TB)染色病理切片及评分,使用酶联免疫吸附测定法(ELISA)检测兔膝关节液白细胞介素-6(IL-6),白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)表达水平,采用免疫组化法检测关节软骨基质金属蛋白酶-13(MMP-13),转化生长因子-β;(TGF-β;),Ⅱ型胶原(Col-Ⅱ)的表达。结果:山药多糖干预5周后,与模型组比较,山药多糖各组关节液中IL-6,IL-1β,TNF-α的含量均明显降低(P<0.05),关节软骨中MMP-13的表达水平均也明显低于模型组(P<0.05),而TGF-β;和Col-Ⅱ的含量明显升高(P<0.05);与山药多糖低、高剂量组比较,山药多糖中剂量组兔膝关节炎症因子IL-6,IL-1β,TNF-α含量明显降低(P<0.05),软骨组织中TGF-β;和Col-Ⅱ的含量明显升高(P<0.05),MMP-13的含量明显降低(P<0.05)。与玻璃酸钠组比较,山药多糖中剂量组兔膝关节炎症因子IL-6,IL-1β,TNF-α含量,软骨组织中TGF-β;,Col-Ⅱ和MMP-13含量差异无统计学意义。结论:山药多糖关节腔注射可明显降低兔膝关节液中IL-6,IL-1β,TNF-α的表达,并能有效抑制关节软骨中MMP-13表达,使关节软骨胶原的降解得到抑制,促进TGF-β;,Col-Ⅱ的合成,对关节软骨起到保护及修复作用,从而延缓关节软骨的退变。
Objective:To investigate the mechanism of joint cavity injection of Dioscoreae Rhizoma polysaccharides (DRP) in protecting against cartilage degeneration and inhibiting the expression of inflammatory factors in the rabbit model of knee osteoarthritis to provide relevant references for the developmen and further research on DRP.Method:Fifty-five New Zealand white rabbits were selected for the induction of knee osteoarthritis model by the modified Hulth’s modeling method.The model rabbits were randomly divided into a model group,a sodium hyaluronate group (1.00 mg·kg^(-1)),and low-(0.7 mg·kg^(-1)),medium-(1.43 mg·kg^(-1)),and high-dose(2.15 mg·kg^(-1))DRP group according to a random number table.One week after modeling,the rabbits in the groups with drug intervention were treated correspondingly for five weeks,once per week,and no intervention was performed in the model group.Five weeks later,the joint specimens were observed by visual observation.The articular cartilage tissues were observed under the light microscope for pathological sections and scores by hematoxylin-eosin(HE)staining and toluidine blue(TB)staining.The expression levels of interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in the synovial fluid were measured by enzyme-linked immunosorbent assay(ELISA),and the expression of matrix metalloproteinase-13(MMP-13),transforming growth factor-β;(TGF-β;),and typeⅡcollagen(Col-Ⅱ)in the articular cartilage were measured by immunohistochemistry.Result:After five weeks of DRP intervention,compared with the model group,the DRP groups exhibited lowered levels of IL-6,IL-1β,and TNF-αin the synovial fluid(P<0.05),reduced expression of MMP-13 in the articular cartilage(P<0.05),and increased levels of TGF-β;and Col-Ⅱ(P<0.05).Compared with the low-dose and high-dose DRP groups,the mediumdose DRP group showed reduced levels of IL-6,IL-1β,and TNF-αin the knee joint(P<0.05),increased levels of TGF-β;and Col-Ⅱin cartilage tissues(P<0.05),and dwindled level of MMP-13(P<
作者
李哲
陈斐斐
韩小康
陈梦晗
杨锋
LI Zhe;CHEN Fei-fei;HAN Xiao-kang;CHEN Meng-han;YANG Feng(Shaanxi University of Chinese Medicine,Xianycwg 712000,China;Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第23期88-96,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81973889)
陕西省重点科技创新团队项目(2013KCT-26)
陕西教育厅服务地方科学研究计划项目(19JC013)。
关键词
膝骨性关节炎
山药多糖
关节腔注射
保护机制
细胞因子
knee osteoarthritis
Dioscoreae Rhizoma polysaccharides
joint cavity injection
protective mechanism
cytokines
