摘要
背景:血管新生是早期膝骨关节炎膝关节微环境的重要病理特征,通过膝关节腔注射microRNA的方式抑制血管新生可能为早期膝关节的治疗提供新的作用靶点。目的:观察miR-20b-5p激动剂对早期膝骨关节炎大鼠关节软骨、软骨下骨血管生成的影响,探讨miR-20b-5p对早期膝骨关节炎大鼠软骨及软骨下骨可能的保护机制。方法:从12只SD雄性大鼠中随机抽取3只作为空白组,余下9只制备早期膝骨关节炎模型后随机分组,即模型组、miR-20b agomir组及miR-20b agomir NC组,每组3只大鼠。造模后1周,空白组不予处理,模型组、miR-20b agomir组、miR-20b agomir NC组分别于双后膝注射0.2 mL生理盐水、miR-20b-5p agomir溶液、miR-20b-5p agomir NC溶液1次。4周后处死各组大鼠取膝关节进行番红O-固绿、免疫组织化学染色及国际骨关节炎研究学会“软骨病理评价系统”评分,实时荧光定量PCR及Western blot检测软骨及软骨下骨中缺氧诱导因子1α、血管内皮生长因子的mRNA和蛋白表达水平。结果与结论:①miR-20b agomir组国际骨关节炎研究学会“软骨病理评价系统”评分明显低于模型组和miR-20b agomir NC组(P<0.05);模型组与miR-20b agomir NC组的国际骨关节炎研究学会“软骨病理评价系统”评分相比差异无显著性意义(P>0.05);②膝关节病理切片显示,相比空白组,其他各组均有软骨基质流失和软骨、软骨下骨的缺氧诱导因子1α、血管内皮生长因子蛋白表达水平增加,其中miR-20b agomir组的软骨基质流失程度以及软骨、软骨下骨的缺氧诱导因子1α、血管内皮生长因子蛋白表达水平低于模型组和miR-20b agomir NC组(P<0.05);③miR-20b agomir组软骨、软骨下骨的缺氧诱导因子1α、血管内皮生长因子的mRNA及蛋白表达水平均低于模型组和miR-20b agomir NC组(P<0.05);④提示过表达miR-20b-5p可下调早期膝骨关节炎软骨及软骨下骨缺氧诱导因子1α、血管内�
BACKGROUND:Angiogenesis is an important pathological feature of early-stage knee osteoarthritis.Intra-articular injection of microRNA for the inhibition of angiogenesis may provide a new target for the treatment of early-stage knee osteoarthritis.OBJECTIVE:To observe the effect of microRNA-20b-5p(miR-20b-5p)agomir on the angiogenesis of articular cartilage and subchondral bone in rats with earlystage knee osteoarthritis,and to explore the possible protective mechanism of miR-20b-5p.METHODS:Twelve male Sprague-Dawley rats were enrolled,three of which were randomly selected as blank group,and the remaining nine rats were randomly divided into model group,miR-20b agomir group and miR-20b agomir NC group,with three rats in each group.One week after modeling,the blank group was not treated,while in the model,miR-20b agomir and miR-20b agomir NC groups,0.2 mL of normal saline,miR-20b-5p agomir saline solution and miR-20b-5p agomir NC saline solution was injected into the rat bilateral knee joints,respectively.Four weeks later,the rats in each group were killed and the knee joints were taken for saffron O-fast green staining,immunohistochemical staining,and Osteoarthritis Research Society International scoring.Real-time fluorescence quantitative PCR and western blot assay were used to detect the expression of hypoxia inducible factor 1αand vascular endothelial growth factor at mRNA and protein levels in the cartilage and subchondral bone.RESULTS AND CONCLUSION:The score of Osteoarthritis Research Society International in the miR-20b agomir group was significantly lower than that in the model group and miR-20b agomir NC group(P<0.05),but there was no significant difference between model group and miR-20b agomir NC group(P>0.05).The pathological sections of the knee joint showed that there was an increase in the loss of cartilage matrix and the expression of hypoxia inducible factor-1αand vascular endothelial growth factor proteins in the cartilage and subchondral bone in all intervention groups compared with the blank
作者
卢启贵
谢平金
罗臻
李飞龙
陈群群
柴生颋
Lu Qigui;Xie Pingjin;Luo Zhen;Li Feilong;Chen Qunqun;Chai Shengting(Shenzhen Hospital,Shanghai University of Traditional Chinese Medicine/Shenzhen Luohu District Hospital of Traditional Chinese Medicine,Shenzhen 518000,Guangdong Province,China;Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China;The Research Center of Basic Integrative Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2022年第29期4658-4665,共8页
Chinese Journal of Tissue Engineering Research
基金
深圳市科创委2020年基础研究面上项目(JCYJ20190812170815559),项目负责人:卢启贵
广东省中医药管理局科研项目(20202085),项目负责人:李飞龙
广东省自然科学基金博士启动纵向协同项目(2018A030310606),项目负责人:陈群群。
