CP-25通过抑制GRK2活性改善小鼠骨关节炎的机制  

CP-25 inhibits GRK2 activity to treat osteoarthritis in mice

作　　者：王惠敏 王旭蕾 储柱平 郭婷婷 徐诺 王康 李颖 魏伟[1] 严尚学[1,2] WANG Hui-min;WANG Xu-lei;CHU Zhu-ping;GUO Ting-ting;XU Nuo;WANG Kang;LI Ying;WEI Wei;YAN Shang-xue(Institute of Clinical Pharmacology,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immuno Drugs,Anhui Medical University;Experimental Animal Center,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,安徽省抗炎免疫药物协同创新中心,安徽合肥230032 [2]安徽医科大学实验动物中心,安徽合肥230032

出　　处：《中国药理学通报》2024年第5期936-944,共9页Chinese Pharmacological Bulletin

基　　金：安徽高校自然科学研究重大项目(No KJ2020ZD15)。

摘　　要：目的研究芍药苷-6-氧-苯磺酸酯(CP-25)通过抑制GRK2活性对骨关节炎(osteoarthritis,OA)小鼠膝关节软骨的保护作用。方法内侧半月板失稳(destabilization of the medial meniscus,DMM)手术诱导构建小鼠骨关节炎模型,实验分为假手术组、模型组、CP-25给药组和帕罗西汀给药组。术后开始灌胃给药。给药12周处死动物,Micro-CT成像观察膝关节软骨退变、骨重塑异常等情况,番红固绿染色观察小鼠关节组织病理,免疫组化、免疫荧光检测软骨组织相关分子表达水平的影响。Western blot检测CP-25用药后软骨细胞的膜蛋白及总蛋白表达水平。结果模型小鼠关节软骨严重退变。CP-25可显著降低关节软骨骨赘数量及软骨下板厚度,促进软骨基质再生,减少软骨基质降解蛋白表达,对膝关节软骨有明显的保护作用。免疫组化和免疫荧光结果显示,CP-25治疗可显著降低膝关节组织中GRK2、ADAMTS5、MMP13的表达,并且升高膝关节组织中ColⅡ、Aggrecan表达。体外实验结果表明,CP-25给药可以显著降低GRK2的膜蛋白及总蛋白表达水平,升高EP4膜蛋白水平,降低MMP13水平。结论CP-25给药可显著促进OA小鼠关节软骨基质再生,减少软骨基质降解,对OA具有治疗作用,其机制与抑制GRK2介导的软骨基质代谢有关。Aim To investigate the protective potential of paeoniflorin 6-oxy-benzenesulfonate(CP-25)in preserving knee cartilage integrity in osteoarthritis mice through inhibition of GRK2 activity.Methods The posttraumatic osteoarthritis model was established following DMM surgery.The experiment consisted of a sham operation group,a model group,a CP-25 administration group,and a paroxetine positive control group.Intragastric administration commenced after the surgery.After 12 weeks of administration,the animals were euthanized.Micro-CT imaging was used to observe the knee cartilage degeneration and abnormal bone remodeling,and the joint histopathology of mice was observed by staining with ferrubens solid green.Immunohistochemistry and immunofluorescence were used to detect the expression level of related molecules in cartilage tissue.Furthermore,Western blot was employed to determine GRK2 and EP4 membrane protein expression levels as well as total protein levels of GRK2 and MMP13 following CP-25 treatment.Results Compared with the sham operation group,the articular cartilage in the model group was significantly degraded,with the cartilage surface calcifying and osteophytes forming.CP-25 could significantly reduce the number of osteophytes and the thickness of the subchondral plate of articular cartilage,promote the regeneration of the cartilage matrix,reduce the expression of cartilage matrix degradation protein,and have a significant protective effect on knee cartilage.Immunohistochemical and immunofluorescence results showed that compared with the model group,CP-25 treatment significantly decreased the expressions of GRK2,ADAMTS5 and MMP13 in knee tissue and increased the expressions of Col II and Aggrecan in knee tissue.The results of in vitro experiments showed that CP-25 administration could significantly reduce the expression levels of GRK2 membrane protein and total protein,increase the level of EP4 membrane protein,and decrease the level of MMP13.Conclusions The administration of CP-25 can significantly promote the

关 键 词：骨关节炎 软骨保护 CP-25 软骨细胞 GRK2 DMM手术 

分 类 号：R-332[医药卫生] R284.1R322.71R345.57R684.3