龙胆苦苷对骨性关节炎软骨细胞外基质的影响  

Effect of Gentiopicroside on Extracellular Matrix of Cartilage Cells in Osteoarthritis

作　　者：刘平举 唐魁韩 孙立[3] 陈在飞 LIU Pingju;TANG Kuihan;SUN Li;CHEN Zaifei(Zunyi Hospital of Traditional Chinese Medicine,Zunyi 563000,China;Guizhou Orthopedic Hospital,Guiyang 550007,China;Guizhou Provincial People′s Hospital,Guiyang 550002,China;Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]遵义市中医院,遵义563000 [2]贵州省骨科医院,贵阳550007 [3]贵州省人民医院,贵阳550002 [4]贵州中医药大学,贵阳550001

出　　处：《世界中医药》2024年第7期957-961,共5页World Chinese Medicine

基　　金：国家自然科学基金项目(81960401);贵州省中医药管理局中医药、民族医药科学技术课题(QZYY-2020-029)。

摘　　要：目的:观察龙胆苦苷对骨性关节炎软骨细胞外基质保护作用的机制。方法:从关节软骨中提取软骨细胞,体外培养传至原代。随机分为空白对照组、模型组和观察组。空白对照组采用常规培养基,模型组在培养基中添加白细胞介素-1β(IL-1β)诱导构建细胞水平关节炎模型,观察组在模型组的基础上添加龙胆苦苷进行干预,在体外共培养72 h后提取RNA。采用实时荧光定量逆转录聚合酶链反应(RT-PCR)等方法在基因水平检测关节软细胞外基质特异性基因Ⅱ型胶原A1、蛋白聚糖和降解酶基质金属蛋白酶-13(MMP-13)、血管性血友病因子裂解蛋白酶-5(ADAMTS-5)基因表达变化。结果:与空白对照组比较,模型组软骨细胞Ⅱ胶原蛋白A1、蛋白聚糖的表达降低,差异均有统计学意义(均P<0.01),MMP-13、ADAMTS-5表达明显增加,差异均有统计学意义(均P<0.01)。与模型组比较,观察组中软骨细胞Ⅱ胶原蛋白A1、蛋白聚糖的表达均出现增加的趋势,其中Ⅱ胶原蛋白A1差异有统计学意义(P<0.05),MMP-13和ADAMTS-5表达减少,差异均有统计学意义(均P<0.05)。结论:龙胆苦苷可上调软骨细胞外基质中Ⅱ胶原蛋白A1、蛋白聚糖的mRNA表达,对骨关节炎的软骨细胞具有保护作用,其作用机制主要是抑制MMP-13的生成。Objective:To observe the protective effect of gentiopicroside on cartilage extracellular matrix in osteoarthritis and explore its mechanism.Methods:Cartilage cells were extracted from articular cartilage and cultured in vitro until primary generation.The experiment set up blank control group,model group and observation group.The control group used conventional culture medium.The model group had interleukin-1β(IL-1β)added to the culture medium to induce an osteoarthritis model at the cellular level.The observation group received gentiopicroside intervention on the basis of the model group conditions.RNA was extracted after 72 h of co-culture in vitro.Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression changes of specific genes for articular cartilage extracellular matrix,such as typeⅡcollagen A1,proteoglycan,matrix metalloproteinase-13(MMP-13),and a disintegrin and metalloproteinase with thrombospondin motifs-5.(ADAMTS-5.)Results:Compared with the control group,the expression of typeⅡcollagen A1 and proteoglycan in the model group decreased significantly(P<0.01),and the expressions of MMP-13 and ADAMTS-5 increased significantly(P<0.01).Compared with the model group,the expressions of typeⅡcollagen A1 and proteoglycan in the observation group increased,with the difference in typeⅡcollagen A1 being statistically significant(P<0.05),and the expressions of MMP-13 and ADAMTS-5 decreased,with statistically significant differences(P<0.05).Conclusion:Gentiopicroside can upregulate the mRNA expression of typeⅡcollagen A1 and proteoglycan in the extracellular matrix of cartilage,and has a protective effect on the cartilage cells in osteoarthritis,primarily by inhibiting the production of MMP-13.

关 键 词：龙胆苦苷 秦艽 关节炎 关节软骨 软骨细胞 基质金属蛋白酶-13 Ⅱ型胶原A1 蛋白聚糖 

分 类 号：R284[医药卫生—中药学] R684.3[医药卫生—中医学]