HSYA对骨关节炎大鼠膝关节软骨损伤的修复作用研究

Study on the repair effect of HSYA on knee cartilage injury in rats with osteoarthritis

目的探讨羟基红花黄色素A(HSYA)对骨关节炎(OA)大鼠膝关节及MMP-3、TGF-β1、Notch1的影响。方法将25只大鼠随机分为正常组(假手术)、OA组(模型大鼠)、低HSYA组(模型大鼠尾部注射2.5 mg/kg HSYA)、中HSYA组(模型大鼠尾部注射5.0 mg/kg HSYA)、高HSYA组(模型大鼠尾部注射10.0 mg/kg HSYA)。采用ELISA检测血清指标IL-1β、TNF-α,Pelletier评分评估膝关节软骨损伤情况,HE染色观察组织病理形态并以Mankin评分进行评估,免疫组化检测MMP-3阳性表达,Western blot检测TGF-β1、Notch1蛋白水平。结果OA组血清中IL-1β、TNF-α水平高于其余各组(P<0.05),高HSYA组IL-1β、TNF-α水平低于低HSYA组和中HSYA组(P<0.05)。各组大鼠Pelletier评分组间比较差异有统计学意义(P<0.05)。OA组软骨和滑膜组织Mankin评分高于其余各组(P<0.05),高HSYA组Mankin评分低于低HSYA组、中HSYA组(P<0.05)。各组大鼠关节组织MMP-3阳性细胞率组间比较差异有统计学意义(P<0.05)。各组大鼠关节组织TGF-β1、Notch1蛋白水平组间比较差异有统计学意义(P<0.05)。结论HSYA能够降低OA大鼠血清炎性因子IL-1β、TNF-α水平,缓解滑膜组织损伤,且存在浓度依赖性,其机制与抑制MMP-3和Notch1表达、提高TGF-β1水平相关。

Objective To investigate the effect of hydroxy safflower yellow A(HSYA)on knee joints and MMP-3,TGF-β1 and Notch1 in osteoarthritis(OA)rats.Methods A total of 25 rats were randomly divided into the normal group(sham operation),the OA group(model rats),the low HSYA group(model rats with tail injection of 2.5 mg/kg HSYA),the medium HSYA group(model rats with tail injection of 5.0 mg/kg HSYA)and the high HSYA group(model rats with tail injection of 10.0 mg/kg HSYA).Serum indicators of IL-1βand TNF-αwere detected by ELISA and the articular cartilage injury was evaluated by Pelletier score.Tissue pathomorphology was observed by HE staining and evaluated by Mankin score.MMP-3 positive expression was measured by immunohistochemical method.The protein levels of TGF-β1 and Notch1 were detected by Western blot.Results The levels of serum IL-1βand TNF-αin the OA group were higher than those in other groups(P<0.05).The levels of serum IL-1βand TNF-αin the high HSYA group were lower than those in the low HSYA group and the medium HSYA group(P<0.05).There were statistically significant differences in Pelletier scores of rats among all groups(P<0.05).Mankin score of cartilage and synovial tissue in the OA group was higher than that in other groups(P<0.05),and Mankin score in the high HSYA group was lower than that in the low HSYA group and the medium HSYA group(P<0.05).There were statistically significant differences in MMP-3 positive expression of the rat joint tissues among all groups.There were statistically significant differences in the protein levels of TGF-β1 and Notch1 of the joint tissues among all groups(P<0.05).Conclusion HSYA can reduce the level of serum inflammatory factors of IL-1βand TNF-αin OA rats and alleviate the injury of synovial tissues in a concentration dependent manner,which is related to inhibiting the expression of MMP-3 and Notch1 and increasing the level of TGF-β1.