健腰密骨片延缓OPG基因敲除小鼠椎间盘退变的研究  

Study of Jianyaomigu Tablet for Delaying IntervertebralDisc Degeneration in OPG Knockout Mice

作　　者：薛纯纯[1,2] 王拥军[2,3] 梁倩倩[2,3] 施杞[2,3] 李晓锋[1,2] XUE Chunchun;WANG Yongjun;LIANG Qianqian;SHI Qi;LI Xiaofeng(Shanghai Municipal Hospital of Traditional Chinese Medicine Affiliated of Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Institute of Spinal Diseases of Shanghai University of Traditional Chinese Medicine,Shanghai 200023,China;Longhua Hospital Affiliated of Shanghai University of Traditional Chinese Medicine,Shanghai 200023,China.)

机构地区：[1]上海中医药大学附属市中医医院,上海200071 [2]上海中医药大学脊柱病研究所 [3]上海中医药大学附属龙华医院

出　　处：《中国中医骨伤科杂志》2021年第8期8-11,共4页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家自然科学基金项目(81973881,81804122);上海市卫生计生系统优秀青年人才培养项目(2018YQ27);上海高校青年骨干教师国内访问学者计划上海市“医苑新星”青年医学人才培养资助计划(沪卫计人事〔2019〕72);上海中医药大学“杏林百人”计划(上中医人字〔2020〕23);上海中医药大学后备卓越中医人才(上中医医管字〔2020〕10)。

摘　　要：目的:观察健腰密骨片对OPG基因敲除小鼠椎间盘退变的影响。方法:30只12周龄小鼠随机分为WT,OPG^(-/-),健腰密骨片组,每组10只,健腰密骨片组采用含药饲料干预3个月,3月后取材获得各组颈椎,以颈5-6椎间盘为观察对象,甲苯胺蓝染色观察椎间盘形态,免疫组化染色观察Ⅰ、Ⅱ、Ⅹ型胶原的表达变化,qPCR分析椎间盘组织中Aggrecan和Col10α1的表达。结果:甲苯胺蓝染色显示健腰密骨片组椎间盘软骨终板中部分缺损和钙化明显减少。免疫组化显示健腰密骨片促进OPG^(-/-)小鼠椎间盘Ⅱ型胶原的表达,降低Ⅹ、Ⅰ型胶原的表达,qPCR结果显示健腰密骨片上调OPG^(-/-)小鼠椎间盘Aggrecan mRNA的表达水平,下调Col10α1 mRNA水平,差异有统计学意义(P<0.05)。结论:健腰密骨片可能通过改善椎间盘软骨细胞代谢延缓OPG^(-/-)小鼠的椎间盘退变。Objective:To observe the effect of Jianyaomigu tablet(JYMGT)on intervertebral disc degeneration in osteoprotegerin(OPG)-deficient mice.Methods:Thirty 12-weeks-old mice were randomly divided into WT,OPG-/- and JYMGT group(n=10).In JYMGT group,OPG-/- mice were administrated with medicated drug diet for 3 months,after 3 months,the cervical intervertebral disc were collected from each group for analyses including toluidine blue staining for morphological,immunostaining for typeⅠ,Ⅱ,Ⅹ collagen changes.The intervertebral disc were also subjected to qPCR for mRNA expression of genes for chondrocytes-Aggrecan,Col10α1.Results:JYMGT could reduced the ossification and defect in the endplate of the intervertebral discs in OPG-/- mice.Immunohisto-chemistry showed that JYMGT promoted expression of type Ⅱ collagen,and decreased the expression of typeⅩ and Ⅰ collagen.In addition,qPCR results investigated that JYMGT increased the expression of Aggrecan and decreased Col10α1 mRNA in the cartilage matrix of intervertebral disc in OPG-/- mice.Conclusion:JYMGT may delay the intervertebral disc degeneration in OPG-/- mice by regulating the chondrocytes metabolism of intervertebral disc.

关 键 词：健腰密骨片 椎间盘退变 骨保护素 软骨细胞 

分 类 号：R-33[医药卫生]