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龙胆苦苷对骨性关节炎软骨细胞外基质的影响

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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.
Effect of Gentiopicroside on Extracellular Matrix of Cartilage Cells in Osteoarthritis
Objective:To observe the protective effect of gentiopicroside on cartilage extracellular matrix in osteoarthritis and ex-plore 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 medi-um.The model group had interleukin-1 β(IL-1 β)added to the culture medium to induce an osteoarthritis model at the cellular lev-el.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 type Ⅱ collagen A1,proteogly-can,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 type Ⅱ collagen A1 and proteoglycan in the model group decreased sig-nificantly(P<0.01),and the expressions of MMP-13 and ADAMTS-5 increased significantly(P<0.01).Compared with the mod-el group,the expressions of type Ⅱ collagen A1 and proteoglycan in the observation group increased,with the difference in type Ⅱcollagen 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 type Ⅱ collagen 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.

GentiopicrosideGentiana macrophyllaArthritisArticular CartilageCartilage CellsMatrix Metalloproteinase-13Type Ⅱ Collagen A1Proteoglycan

刘平举、唐魁韩、孙立、陈在飞

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遵义市中医院,遵义,563000

贵州省骨科医院,贵阳,550007

贵州省人民医院,贵阳,550002

贵州中医药大学,贵阳,550001

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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

国家自然科学基金贵州省中医药局中医药、民族医药科学技术课题

81960401QZYY-2020-029

2024

世界中医药
世界中医药学会联合会

世界中医药

CSTPCDCHSSCD北大核心
影响因子:1.266
ISSN:1673-7202
年,卷(期):2024.19(7)