骨健口服液联合间充质干细胞来源外泌体治疗小鼠膝骨关节炎的实验研究  

Gujian(骨健)peroral liquids combined with mesenchymal stem cell-derived exosome against knee osteoarthritis in mice:an experimental study

作　　者：吴捷 张若谷 邱敏 程良艳 徐建波 金敏伟[2] 金红婷[2] 王萍儿 WU Jie;ZHANG Ruogu;QIU Min;CHENG Liangyan;XU Jianbo;JIN Minwei;JIN Hongting;WANG Pinger(The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China;Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China)

机构地区：[1]浙江中医药大学第一临床医学院,浙江杭州310053 [2]浙江省中医院,浙江杭州310006

出　　处：《中医正骨》2024年第4期28-36,共9页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：浙江省中医药科技计划项目(2022ZB116)。

摘　　要：目的:探讨骨健口服液联合间充质干细胞来源外泌体(mesenchymal stem cell-derived exosome,MSC-EXO)治疗小鼠膝骨关节炎(knee osteoarthritis,KOA)的效果及作用机制。方法:将25只12周龄雄性无特定病原C57BL/6J小鼠随机分为假手术组、模型组、骨健组、MSC-EXO组、骨健MSC-EXO组,每组5只。假手术组仅切开关节囊而不切断内侧半月板,其余4组均采用内侧半月板失稳法构建KOA模型。骨健组、骨健MSC-EXO组小鼠每日用0.2 mL的骨健口服液灌胃,假手术组、模型组、MSC-EXO组小鼠每日用等量的生理盐水灌胃,均连续8周。MSC-EXO组、骨健MSC-EXO组小鼠每周膝关节腔注射1次10μL的MSC-EXO,其余各组小鼠每周膝关节腔注射1次等量的磷酸盐缓冲液(phosphate buffer solution,PBS),均连续8周。干预8周后处死小鼠,进行小鼠膝关节Micro-CT分析、膝关节软骨组织病理学观察、膝关节软骨组织中Ⅱ型胶原蛋白α1链(collagenⅡα1,Col2A1)表达水平检测。结果:①小鼠膝关节Micro-CT分析结果。假手术组、骨健组、MSC-EXO组、骨健MSC-EXO组小鼠的骨体积分数均低于模型组(P=0.004,P=0.007,P=0.040,P=0.001),骨健组、MSC-EXO组小鼠的骨体积分数与骨健MSC-EXO组的差异均无统计学意义(P=0.108,P=0.126)。假手术组、骨健组、MSC-EXO组、骨健MSC-EXO组小鼠的骨小梁厚度均大于模型组(P=0.015,P=0.023,P=0.023,P=0.009),骨健组、MSC-EXO组小鼠的骨小梁厚度与骨健MSC-EXO组的差异均无统计学意义(P=0.347,P=0.062)。假手术组和骨健组小鼠的骨小梁数量多于模型组(P=0.016,P=0.021),MSC-EXO组、骨健MSC-EXO组小鼠的骨小梁数量与模型组的差异均无统计学意义(P=0.196,P=0.206)。假手术组、骨健组、MSC-EXO组、骨健MSC-EXO组小鼠的骨小梁分离度均小于模型组(P=0.016,P=0.012,P=0.007,P=0.001),骨健MSC-EXO组小鼠的骨小梁分离度小于骨健组(P=0.016),MSC-EXO组小鼠的骨小梁分离度与骨健MSC-EXO组的差异无统计学�Objective:To observe the effects of Gujian(骨健,GJ)peroral liquids combined with mesenchymal stem cell-derived exosome(MSC-EXO)on knee osteoarthritis(KOA)in mice,and to explore its mechanism.Methods:Twenty-five 12-week-old specific pathogen free(SPF)-grade male C57BL/6J mice were randomized into sham-operated group,model group,GJ group,MSC-EXO group,GJ combined MSC-EXO group,5 cases in each group.All mice but the ones in sham-operated group were subjected to destabilized medial meniscus(DMM)method on the right knees for inducing KOA,while the ones in sham-operated group underwent surgery for merely incising the joint capsule,but without transecting medial meniscus.After successful modeling,the mice in GJ group and GJ combined MSC-EXO group were intervened by intragastric administration with 0.2 mL GJ peroral liquids,while the ones in sham-operated group,model group and MSC-EXO group with the same dose of normal saline,once a day for consecutive 8 weeks;moreover,the mice in MSC-EXO group and GJ combined MSC-EXO group were further intervened by knee intra-articular injection of 10μL MSC-EXO,while the ones in sham-operated group,model group and GJ group by the same dose of phosphate buffer solution(PBS),once a week for consecutive 8 weeks.After the end of intervention,the mice were sacrificed by cervical dislocation method,and their right knees were harvested for scanning and analysing by using Micro-CT,followed by observation on histopathological changes of knee tissues and determination of the expression level of collagenⅡα1(Col2A1)in knee cartilage tissues.Results:①The bone volume fraction was lower in sham-operated group,GJ group,MSC-EXO group and GJ combined MSC-EXO group compared to model group(P=0.004,P=0.007,P=0.040,P=0.001),while,there was no statistical difference between GJ group and GJ combined MSC-EXO group as well as between MSC-EXO group and GJ combined MSC-EXO group(P=0.108,P=0.126).The trabecular thickness was greater in sham-operated group,GJ group,MSC-EXO group and GJ combined MSC-EXO grou

关 键 词：骨关节炎 膝 小鼠 间质干细胞 外泌体 骨健口服液 软骨 动物实验 

分 类 号：R285.5[医药卫生—中药学]