蛇床子素对Ⅱ型胶原蛋白诱导的关节炎大鼠软骨损伤的改善及免疫调节作用
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江苏省科技项目基础研究计划项目(BK20181459)


Improvement of cartilage damage and immunomodulatory effects of osthole on type Ⅱ collagen⁃induced arthritis rats
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    摘要:

    目的:研究蛇床子素对Ⅱ型胶原蛋白诱导的关节炎大鼠软骨损伤的改善及免疫调节作用。方法:用牛Ⅱ型胶原诱导构建类风湿性关节炎大鼠模型,在造模完成1周后以不同浓度(10、20、40 mg/kg)蛇床子素灌胃给药。每4 d测定大鼠的爪子体积并评定关节炎指数。30 d后,称重测定脾脏指数和胸腺指数,HE染色观察软骨损伤情况,ELISA检测白细胞介素(IL)-1β、IL-6和肿瘤坏死因子α(TNF-α)含量,qRT-PCR检测基质金属蛋白酶(MMP)-1、MMP-3和MMP-13 mRNA表达,Western blot检测Ki67、PCNA、Caspase-3、Caspase-9、MMP-1、MMP-3、MMP-13、Wnt1、β-catenin和LRP5表达。结果:与模型组相比,各给药组大鼠的爪子体积、关节炎指数、脾脏指数和胸腺指数均降低,软骨细胞排列紊乱程度减轻,炎性细胞数量减少,Ki67和PCNA表达均上调,Caspase-3和Caspase-9表达均下调,IL-1β、IL-6和TNF-α 含量降低,MMP-1、MMP-3、MMP-13、Wnt 1、β-catenin和LRP5表达均下调,其中以40 mg/kg组的作用最显著(P < 0.01)。结论:蛇床子素能够减轻类风湿性关节炎大鼠的软骨损伤、炎症反应及关节软骨基质胶原的破坏,该作用可能是通过抑制Wnt信号通路的激活来实现的。

    Abstract:

    Objective:To investigate the improvement of cartilage damage and immunomodulatory effects of osthole on type Ⅱ collagen-induced arthritis rats. Methods:A rheumatoid arthritis rat model was inducd by bovine type Ⅱ collagen. Osthole was administered by intragastric injection one week after the modeling was established. The paw volume of each group and the arthritis index were evaluated every 4 days. After 30 days,the spleen index and thymus index were determined by weighing method. Cartilage damage was observed by HE staining. The levels of IL-1β,IL-6 and TNF-α in peripheral blood were measured by ELISA. The expressions of MMP-1,MMP-3 and MMP-13 mRNA were determined by qRT-PCR. The levels of Ki67,PCNA,Caspase-3,Caspase-9,MMP-1,MMP-3,MMP-13,Wnt 1,β-catenin and LRP5 were detected by Western blot. Results:Compared with the model group,the paw volume,arthritis score,spleen index and thymus index of the osthole treated group were decreased,the degree of chondrocyte arrangement disorder and inflammatory cells were reduced,the expressions of Ki67 and PCNA were up-regulated,and Caspase-3 and Caspase-9 were down-regulated,the contents of IL-1β,IL-6 and TNF-α were decreased,the expressions of MMP-1,MMP-3,MMP-13,Wnt 1,β-catenin and LRP5 were down-regulated,and the effects were greatest in the Ost 40 mg/kg group(P < 0.01). Conclusion:Osthole can alleviate cartilage damage,inflammation,and destruction of collagen in the articular cartilage matrix of rats with rheumatoid arthritis,which may be achieved by inhibiting the activation of the Wnt signaling pathway.

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贝宇飞,朱 清,王 静,范震宇,殷晓芹.蛇床子素对Ⅱ型胶原蛋白诱导的关节炎大鼠软骨损伤的改善及免疫调节作用[J].南京医科大学学报(自然科学版),2020,(4):521-527

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  • 收稿日期:2019-09-29
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  • 在线发布日期: 2020-05-08
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