摘要
[目的]探讨miR-145体内转染对骨性关节炎模型小鼠关节软骨的影响。[方法]8周龄雄性C57小鼠40只,随机分为4组,每组10只。分别为正常对照组(normal control,NC)、骨性关节炎模型组(osteoarthritis model,OAM)、正常动物转染组(transferred normal animal,TNA)及OA模型转染组(transferred OAM,TOAM)。对OAM和TOAM组动物切断全部内侧半月板-胫骨韧带建立OA模型。7、10 d时,对TOAM组和TNA组关节腔内注射载有miR-145的腺病毒液20μl。再过4 d后过度麻醉处死动物。行大体和组织学观察,并采用qPCR和Western blot检测相应标志物。[结果]形态方面,与NC组相比,OAM组关节软骨结构明显紊乱,纤维细胞增生,基质着色主要呈现蓝绿色。TNA组的软骨结构完整,但陷窝中出现空泡,基质着色呈蓝绿色。相比之下,TOAM组软骨结构完整,关节软骨基质着色呈红色,与NC组接近。免疫组化检测显示,与NC组相比,OAM组和TNA组的FRS2、LC3和p62染色的OD值均显著增加(P<0.05);而TOAM组的FRS2、LC3和p62染色的OD值显著低于OAM组和TNA组(P>0.05)。定量检测方面,与NC组相比,OAM组的miRNA-145的mRNA表达水平显著下降(P<0.05),而体内转染后,TNA组和TOAM组的miRNA-145的mRNA表达水平显著高于NC组(P<0.05)。OAM组的LC3II/I比值降低且p62表达水平显著升高(P<0.05)。TOAM组的p62的表达水平显著低于OAM组和TNA组(P<0.05)。[结论]miR-145体内转染可减轻关节失稳状的软骨退变,其机理可能与改变软骨自噬功能有关。
[Objective]To investigate the effect of miR-145 transfection in vivo on articular cartilage of osteoarthritis(OA)model in mice.[Methods]Forty male C57 mice aged 8 weeks were randomly divided into 4 groups with 10 mice in each group,including normal control group(NC),osteoarthritis model group(OAM),transfected normal animal(TNA),and transfected OA model(TOAM).The animals in the OAM and TOAM groups received total resection of medial meniscus-tibial ligament to create OA model.At 7 d and 10 d,20μl of adenosis vector carrying miR-145 was injected into the joint cavity of the mice in the TOAM group and TNA group.The animals were sacrificed after another 4 days by excessive anesthesia,and then gross and histological observation were conducted,while qPCR and western blot assays were used to detect corresponding markers.[Results]In terms of morphology,the structure of articular cartilage in OAM group was obviously disordered,with fibrocyte proliferation and matrix stained in mainly blue-green compared with the NC group,while which preserved intact in structure,but with vacuoles in the lacunae,and the matrix in blue-green in the TNA group,whereas intact cartilage structure was noted in the TOAM group with red color of articular cartilage matrix similar to the NC group.In term of immunohistochemistry,the OD values of FRS2,LC3 and p62 staining were significantly increased in OAM group compared with those in the NC group and TNA group(P<0.05),whereas which in the TOAM group were significantly lower than those in OAM group and TNA group(P<0.05).In terms of the quantitative detections,m RNA expression level of miR-145 in OAM group was significantly decreased(P<0.05),while which significantly ramped up in the TNA group and TOAM group compared with NC group(P<0.05).In OAM group the LC3II/I ratio significantly decreased(P<0.05)and p62 expression level were significantly increased compared with those in NC group(P<0.05),while which in the TOAM group was significantly lower than that in OAM group and TNA group(P<0.05).[Conclusion]
作者
苑龙
郑海艺
李森
卞继超
田珂
王国栋
YUAN Long;ZHENG Hai-yi;LI Sen;BIAN Ji-chao;TIAN Ke;WANG Guo-dong(College of Clinical Medicine,Jining Medical University,Jining 272000,China;Department of Osteoarthrosis,Affiliated Hospital,Jining Medical University,Jining,Shandong 272000,China)
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2022年第18期1674-1679,共6页
Orthopedic Journal of China
基金
国家自然科学基金项目(编号:81871814)
济宁市重点研发计划项目(编号:2020YXNS022)。
