落新妇苷通过抗炎和软骨基质降解对膝骨性关节炎小鼠的治疗作用研究

The therapeutic effect of astilbin on knee osteoarthritis in mice through anti-inflammatory and cartilage matrix degradation

目的:探讨落新妇苷对小鼠膝骨性关节炎(OA)的治疗作用和机制。方法:将60只C57BL/6小鼠随机分为假手术组、模型组、落新妇苷低剂量(5 mg/kg)组、中剂量(10 mg/kg)组、高剂量(20 mg/kg)组和阳性药组,每组10只。除假手术组外,其他各组采用内侧半月板失稳术建立OA小鼠模型。治疗4周后,测定各组关节炎评分、关节肿胀度和机械性痛阈值。测定血清白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)、前列腺素E2(PGE2)含量及组织基质金属酶-3(MMP-3)、MMP-13、含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-4(ADAMTS-4)和一氧化氮(NO)含量。Western blotting法测量软骨组织中诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2)蛋白表达水平。苏木精—伊红(HE)和番红O染色观察软骨损伤和基质降解。结果:与模型组比较,落新妇苷各剂量组和阳性药组小鼠的关节炎评分和关节肿胀度明显减小,机械性痛阈值明显增加,血清中IL-1β、IL-6、TNF-α和PGE2,以及组织中NO、MMP-3、MMP-13和ADAMTS-4含量减小(均P<0.05);落新妇苷各剂量组小鼠软骨组织中iNOS和COX-2蛋白表达下调,软骨组织OARSI评分和钙化软骨(CC)减小、透明软骨(HC)厚度增加(均P<0.05)。结论:落新妇苷可有效改善OA小鼠症状,可能与抗炎和软骨基质降解有关。

Objective:To investigate the therapeutic effect and mechanism of astilbin on knee osteoarthritis(OA)in mice.Methods:A total of 60 C57 BL/6 mice were randomly divided into sham-operation group,model group,astilbin low-dose group,medium-dose group,high-dose group and positive drug group,with 10 rats in each group.A mouse model of osteoarthritis was established by medial meniscus instability,and drugs were given for4 weeks.The arthritis score,joint swelling degree and mechanical pain threshold were measured.The levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α) and prostaglandin E2(PGE2) in serum and the contents of matrix metallogenase-3(MMP-3),MMP-13,a disintegrin and metalloproteinase with thrombospondin motifs-4(ADAMTS-4) and nitric oxide(NO) in tissues were detected.Western blotting was used to measure the protein expressions of inducible nitric oxide synthase(iNOS) and cyclooxyganese-2(COX-2) in cartilage tissues.Hematoxylin-eosin and Safranin O staining was used to observe the cartilage damage and matrix degradation.Results:Compared with the model group,the arthritis score,joint swelling,the levels of IL-1β,IL-6,TNF-α,PGE2,NO,MMP-3,MMP-13 and ADAMTS-4,the expressions of iNOS and COX-2,OARSI score and thickness of calcified cartilage tissue in each astilbin dose group and positive drug group were significantly reduced,while the thickness of hyaline cartilage tissue and mechanical pain threshold were significantly increased(P<0.05).Conclusion:Astilbin can effectively improve the symptoms of OA mice,which may be related to the inhibition of inflammation and cartilage matrix degradation.