基于LncRNA GAS5/miR-21调控软骨基质代谢探讨独活寄生汤干预大鼠膝骨关节炎的作用机制

目的观察独活寄生汤对膝骨关节炎(KOA)大鼠软骨LncRNA GAS5/miR-21及其下游MMP-3、MMP-9、MMP-13和ADAMTS-5的影响,探讨独活寄生汤治疗KOA的作用机制。方法2月龄SD大鼠66只,适应性喂养1周后,随机分为正常组(18只)和造模组(48只)。应用改良Hulth法复制大鼠KOA模型,造模2周后,从正常组和造模组中随机抽取3只大鼠进行小动物MRI观察,鉴定造模是否成功。造模成功后,采用抽签法分为模型组、治疗组和西药组各15只。正常组和模型组给予0.9%生理盐水灌胃,治疗组给予独活寄生汤灌胃,西药组给予盐酸氨基葡萄糖胶囊灌胃。治疗12周后,切取右侧胫骨平台软骨组织,Real-time PCR法检测LncRNA GAS5、miRNA-21、MMP-3、MMP-9、MMP-13、ADAMTS-5基因表达。结果Real-time PCR结果显示,与模型组相比,独活寄生汤和盐酸氨基葡萄糖胶囊均能抑制LncRNA GAS5、MMP-3、MMP-9、MMP-13、ADAMTS-5基因表达(P<0.05),促进miR-21基因表达(P<0.05),但2组间无统计学意义(P>0.05)。结论独活寄生汤可能通过调控LncRNA GAS5/miR-21,延缓软骨细胞外基质降解,起到治疗KOA的作用。

Mechanism of Duhuo Jisheng Decoction in Treating Rats with Knee Osteoarthritis Based on LncRNA GAS5/miR-21 Regulating Metabolism of Cartilage Extracellular Matrix

Objective: To explore the mechanism of Duhuo Jisheng Decoction(DHJSD) in the treatment of rats with knee osteoarthritis by observing the effect of DHJSD on LncRNA GAS5/miR-21 and its downstream MMP-3, MMP-9, MMP-13, ADAMTS-5 in cartilage of rats. Methods: After one week of acclimation, two-month-old male SD rats(n=66) were randomly divided into normal group(n=18) and osteoarthritis model group(n=48). The model group was made according to modified Hulth method. Two weeks after operation, three rats from each group were randomly selected for knee joint magnetic resonance imaging(MRI) to determine whether the model was successful. After successfully establish the model, the rats were randomly divided into three groups: model group, treatment group and control group, with 15 rats in each group. The normal group and the model group were given 0.9% NaCl solution, the treatment group was given DHJSD, and the control group was given glucosamine hydrochloride capsule for 12 weeks via gavage. The medial tibial plateau cartilage tissue of right knee was removed after the treatment and the gene expression of LncRNA GAS5, mi RNA-21, MMP-3, MMP-9, MMP-13, ADAMTS-5 was detected by Real-time PCR. Results: Real-time PCR showed that compared with the model group, DHJSD and glucosamine hydrochloride capsule both could inhibit the gene expression of LncRNA GAS5, MMP-3, MMP-9, MMP-13, ADAMTS-5 and promote the expression of mi R-21(P<0.05), but there was no significant difference between the two groups(P >0.05). Conclusion: DHJSD may delay degradation of cartilage extracellular matrixby regulating LncRNA GAS5/miR-21, which may explain its clinical efficacy in knee osteoarthritis treatment.