龟鹿二仙胶对膝骨关节炎大鼠软骨细胞IHH-GLI信号通路的影响  

Effects of Guilu Erxian Gum on IHH-GLI signal pathway of chondrocytes in rats with knee osteoarthritis

作　　者：郑珍萍 吴伟欣 李晔 王和鸣 耿秋东 李楠 ZHENG Zhenping;WU Weixin;LI Ye;WANG Heming;GENG Qiudong;LI Nan(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian University of Traditional Chinese Medicine Affiliated Rehabilitation Hospital,Fuzhou 350003,China;Key Laboratory of Orthopedics&Traumatology of Traditional Chinese Medicine and Rehabilitation,Ministry of Education,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中医学院,福州350122 [2]福建中医药大学附属康复医院,福州350003 [3]中医骨伤及运动康复教育部重点实验室,福州350122

出　　处：《中华中医药杂志》2024年第4期1747-1752,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81973880);福建中医药大学校管科研课题(No.X2021022)。

摘　　要：目的:观察龟鹿二仙胶含药血清对白介素-1β(IL-1β)诱导大鼠肥大软骨细胞印度刺猬蛋白-胶质细胞瘤转录因子(IHH-GLI)信号通路的作用,探讨龟鹿二仙胶对膝骨关节炎(KOA)的影响。方法:制备含药血清,提取大鼠软骨细胞,分为空白组、模型组、龟鹿组、抑制剂组,构建大鼠KOA软骨细胞肥大模型,免疫荧光观察GLI入核情况。CCK-8检测含药血清干预最佳时效及量效关系。含药血清干预24 h后,Real-time PCR、Western Blot检测软骨细胞IHH、平滑蛋白(Smo)、Runt相关转录因子2(Runx2)、X型胶原(COL10A1)和基质金属蛋白酶13(MMP-13)的mRNA及蛋白表达情况。结果:荧光显微镜下,模型组GLI信号进入细胞核。与模型组比较,龟鹿组及抑制剂组IHH、Smo、Runx2、COL10A1、MMP-13 mRNA表达显著降低(P<0.01,P<0.05),IHH、Smo、Runx2蛋白表达显著下降(P<0.05,P<0.01)。结论:龟鹿二仙胶调控肥大软骨细胞的分解合成代谢平衡,减少细胞外基质的降解和钙化,改善失稳态环境,可能与其抑制IHH-GLI信号通路有关。Objective:To observe of the effects of Guilu Erxian Gum containing serum on interleukin-1β(IL-1β)inducing the role of IHH-GLI signaling pathway in rat hypertrophic chondrocytes and explore the effects of Guilu Erxian Gum on knee osteoarthritis(KOA).Methods:Drug-containing serum was prepared,and rat chondrocytes were extracted and divided into blank group,model group,Guilu group and inhibitor group.Rat KOA chondrocyte hypertrophy model were constructed and GLI entry into nucleus by immunofluorescence were observed.CCK-8 detected the best time-effect and dose-effect relationship of drug-containing serum intervention.After 24 hours of intervention with serum containing drugs,Real-time PCR and Western Blot were used to detect the mRNA and protein expression of IHH,Smo,Runx2,COL10A1,MMP-13 in chondrocytes.Results:Under fluorescence microscope,GLI signal of model group entered the nucleus.Compared with the model group,the expression of IHH,Smo,Runx2,COL10A1,MMP-13 mRNA and the expression of IHH,Smo and Runx2 protein were significantly decreased(P<0.01,P<0.05)in the Guilu group and the inhibitor group.Conclusion:Guilu Erxian Gum regulates the balance of catabolism and metabolism of hypertrophic chondrocytes,reduces the degradation and calcification of extracellular matrix,and improves the unstable environment,which may be related to its inhibition of IHH-GLI signal pathway.

关 键 词：膝骨关节炎 软骨细胞肥大 龟鹿二仙胶 IHH-GLI信号通路 

分 类 号：R285.5[医药卫生—中药学]