骨髓间充质干细胞对膝关节炎大鼠软骨损伤及KDM6A/SOX9信号通路的影响

Effects of bone marrow mesenchymal stem cells on cartilage damage and KDM6A/SOX9 signaling pathway in rats with knee osteoarthritis

目的观察骨髓间充质干细胞对膝关节骨性关节炎大鼠的影响,并探讨其机制。方法将40只SD大鼠随机分为正常对照组(Control)、膝关节骨性关节炎组(KOA)、骨髓间充质干细胞组(BMSCs),每组10只,剩余10只用于骨髓间充质干细胞的分离。处理4周后进行大鼠关节指数(AI)评分;HE染色观察软骨组织病理学变化;免疫组化检测软骨组织Col II、IL-6、MMP-3、MMP-13表达;ELISA法检测血清IL-6、TNF-α、MMP-3、MMP-13含量;qRT-PCR检测软骨组织Col II、Col I、TNF-α、IL-6、MMP-13、KDM6A、SOX9、Aggrecan mRNA表达;Western blot检测软骨组织KDM6A、SOX9、Aggrecan蛋白表达。结果与Control组比较,KOA组大鼠AI评分、血清IL-6、TNF-α、MMP-3、MMP-13含量、软骨组织IL-6、MMP-3、MMP-13表达明显升高(P<0.05);Col II、KDM6A、Aggrecan、SOX9 mRNA和蛋白表达明显降低(P<0.05)。与KOA组比较,BMSCs组大鼠AI评分,血清IL-6、TNF-α、MMP-3、MMP-13含量、软骨组织IL-6 mRNA表达明显降低(P<0.05),软骨组织KDM6A、Aggrecan mRNA和蛋白表达明显升高(P<0.05)。结论骨髓间充质干细胞明显改善膝关节骨性关节炎病理损伤,可能通过调节KDM6A/SOX9信号通路发挥对膝关节骨性关节炎大鼠的治疗作用。

Objective To observe the effect of bone marrow mesenchymal stem cells on knee osteoarthritis rats and to explore the possible mechanisms.Methods Forty SD rats were randomly divided into normal control group(Control,saline 0.3 mL,once a week for 4 times),knee osteoarthritis group(KOA,saline 0.3 mL,once a week for 4 times),bone marrow mesenchymal stem cells group(BMSCs,0.3 mL,once a week for 4 times),10 rats in each group,and the remaining 10 rats were used for the isolation of bone marrow MSCs.After 4 weeks,the joint index(AI)scores of rats were performed;the pathological changes of cartilage tissue were observed by HE staining;immunohistochemistry was performed to detect the expression of Col II,IL-6,MMP-3 and MMP-13 in cartilage tissues;the content of IL-6,TNF-α,MMP-3 and MMP-13 in serum was detected by ELISA method;and the expression of Col II,Col I,TNF-α,IL-6,MMP-13,KDM6A,SOX9 and Aggrecan mRNA in cartilage tissue was detected by qRT-PCR;Western blot to detect KDM6A,SOX9,Aggrecan protein expression in cartilage tissue.Results Compared with the control group,AI score,serum IL-6,TNF-α,MMP-3,MMP-13 content,and cartilage tissue IL-6,MMP-3,MMP-13 expression were significantly higher in the KOA group rats(P<0.05);Col II,KDM6A,Aggrecan,SOX9 mRNA and protein expression significantly decreased(P<0.05).Compared with the KOA group,the BMSCs group significantly decreased AI score,significantly decreased serum IL-6,TNF-α,MMP-3,MMP-13 levels,and IL-6 mRNA expression in cartilage tissue(P<0.05),and significantly increased KDM6A,Aggrecan mRNA and protein expression in cartilage tissue(P<0.05).Conclusion Bone marrow mesenchymal stem cells significantly improved the pathological damage of knee osteoarthritis and may exert therapeutic effects on knee osteoarthritis rats by regulating the KDM6A/SOX9 signaling pathway.