中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (31): 5002-5008.doi: 10.12307/2023.184

• 骨科植入物相关基础实验 Basic experiments of orthopedic implant • 上一篇    下一篇

黄芪多糖抑制Toll样受体4/核因子κB p65通路治疗大鼠膝骨关节炎

陈景涛,陈  有,李玉静,刘志刚   

  1. 河南大学第一附属医院骨科,河南省开封市  475001
  • 收稿日期:2022-03-17 接受日期:2022-05-24 出版日期:2023-11-08 发布日期:2023-01-31
  • 通讯作者: 刘志刚,硕士,主任医师,河南大学第一附属医院骨科,河南省开封市 475001
  • 作者简介:陈景涛,男,1978年生,河南省新密市人,汉族,2007年郑州大学毕业,硕士,主治医师。
  • 基金资助:
    开封市科技发展计划项目(2003028),项目负责人:陈有

Astragalus polysaccharide inhibits Toll-like receptor 4/nuclear factor kappaB p65 pathway in the treatment of knee osteoarthritis in rats

Chen Jingtao, Chen You, Li Yujing, Liu Zhigang   

  1. Department of Orthopedics, The First Affiliated Hospital of Henan University, Kaifeng 475001, Henan Province, China
  • Received:2022-03-17 Accepted:2022-05-24 Online:2023-11-08 Published:2023-01-31
  • Contact: Liu Zhigang, Master, Chief physician, Department of Orthopedics, The First Affiliated Hospital of Henan University, Kaifeng 475001, Henan Province, China
  • About author:Chen Jingtao, Master, Attending physician, Department of Orthopedics, The First Affiliated Hospital of Henan University, Kaifeng 475001, Henan Province, China
  • Supported by:
    Kaifeng Municipal Science and Technology Development Program, No. 2003028 (to CY)

摘要:


文题释义:

骨关节炎:是临床常见的以关节软骨退行性病变引发一种无菌性炎症性关节疾病,可进展为关节软骨退变、断裂以及缺损,甚至危害整个关节面。此次研究采用木瓜蛋白酶诱导大鼠膝骨关节炎模型。 
黄芪多糖:是中药黄芪中主要的有效生物活性成分,具有抗氧化、抗炎、免疫调节以及抗肿瘤等多种功效,已被广泛用于临床多种疾病的研究。细胞外基质蛋白酶、炎症因子、氧自由基等活性因子在促进骨关节炎发展过程中发挥重要作用,而黄芪多糖对改善细胞外基质、胶原以及氧化应激等均具有一定的作用。

背景:黄芪多糖治疗骨关节炎具有一定的作用,Toll样受体4/核因子κB通路在膝骨关节炎发病机制中发挥重要作用。
目的:探讨黄芪多糖对膝骨关节炎的干预作用及其机制。
方法:40只SD大鼠随机分为对照组、模型组、黄芪多糖低剂量组、黄芪多糖高剂量组和脂多糖组(n=8),除对照组之外,其余各组均建立膝骨关节炎模型。造模成功1周后,黄芪多糖低、高剂量组分别向大鼠膝关节腔内注射质量浓度为0.1,0.5 g/L的黄芪多糖生理盐水溶液
0.2 mL,脂多糖组注射0.5 g/L 黄芪多糖 0.2 mL+500 µg/kg脂多糖 0.2 mL;模型组和对照组同法注射等量生理盐水;均3 d注射 1次,干预6周。干预结束后,苏木精-伊红、番红O-固绿染色观察软骨、滑膜组织病理学变化,采用Mankin’s评分和国际骨关节炎研究协会评分进行评估;ELISA法检测滑膜组织中白细胞介素1β、白细胞介素6、肿瘤坏死因子α、一氧化氮、丙二醛、超氧化物歧化酶表达水平;Western blot检测软骨组织中聚集蛋白聚糖、Ⅱ型胶原、基质金属蛋白酶3及核因子κB p65、Toll样受体4、MyD88表达。

结果与结论:①与模型组相比,黄芪多糖低、高剂量组的Mankin’s评分、国际骨关节炎研究协会评分、白细胞介素1β、白细胞介素6、肿瘤坏死因子α、一氧化氮、丙二醛、基质金属蛋白酶3水平及核因子κB p65、Toll样受体4、MyD88表达均显著降低,聚集蛋白聚糖、Ⅱ型胶原表达及超氧化物歧化酶活性均显著升高(P < 0.05),且黄芪多糖高剂量组均较低剂量组作用更明显(P < 0.05);②与黄芪多糖高剂量组比,脂多糖组Mankin’s评分、国际骨关节炎研究协会评分、白细胞介素1β、白细胞介素6、肿瘤坏死因子α、一氧化氮、丙二醛、基质金属蛋白酶3水平及核因子κB p65、Toll样受体4、MyD88表达均显著升高,聚集蛋白聚糖、Ⅱ型胶原表达及超氧化物歧化酶活性均显著降低(P < 0.05);③提示黄芪多糖可通过抑制Toll样受体4/核因子κB p65通路的激活改善膝骨关节炎的胞外基质降解、炎症反应和氧化应激水平,改善关节损伤。

https://orcid.org/0000-0002-2845-7219(陈景涛) 

中国组织工程研究杂志出版内容重点:人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程

关键词: 黄芪多糖, 膝骨关节炎, 炎症, Toll样受体4, 核因子κB p65, 大鼠

Abstract: BACKGROUND: Astragalus polysaccharide has a certain role in the treatment of arthritis, and abnormal Toll-like receptor 4/nuclear factor κB pathway plays an important role in the pathogenesis of knee osteoarthritis. 
OBJECTIVE: To investigate the effect of astragalus polysaccharide on osteoarthritis and its mechanism. 
METHODS: Forty SD rats were randomly divided into control group (the same volume of normal saline), model group (the same volume of normal saline), astragalus polysaccharide low-dose group (0.2 mL of 0.1 /L astragalus polysaccharide intervention), astragalus polysaccharide high-dose group (0.2 mL of 0.5 g/L astragalus polysaccharide intervention), and lipopolysaccharide group (0.2 mL of 0.5 g/L astragalus polysaccharide+0.2 mL of 500 µg/kg lipopolysaccharide). Interventions in each group began at 1 week after modeling, once every 3 days, for 6 continuous weeks. After intervention, histopathological changes of the cartilage and synovial membrane were observed by hematoxylin-eosin staining and saffron O-solid green staining. Mankin’s score and Osteoarthritis Research Society International score were performed for histological evaluation. The levels of interleukin-1β, interleukin-6, tumor necrosis factor-α, nitric oxide, malondialdehyde, and superoxide dismutase in synovial tissue were detected by ELISA. The protein levels of Aggrecan, collagen type II, matrix metalloproteinase 3, nuclear factor-κB p65, Toll-like receptors 4 and myeloid differentiation factor 88 in cartilage tissue were detected by western blot assay. 
RESULTS AND CONCLUSION: (1) Compared with the model group, the Mankin’s score, Osteoarthritis Research Society International score, the levels of interleukin-1β, interleukin-6, tumor necrosis factor-α, nitric oxide, malondialdehyde, and matrix metalloproteinase 3, and the expressions of nuclear factor-κB p65, Toll-like receptors 4, and myeloid differentiation factor 88 were significantly decreased in the astragalus polysaccharide low- and high-dose groups, while the expression levels of Aggrecan, collagen type II, and superoxide dismutase activity were significantly increased (P < 0.05). Furthermore, the astragalus polysaccharide high-dose group showed better effects than the low-dose group (P < 0.05). (2) Compared with the astragalus polysaccharide high-dose group, the Mankin’s score, Osteoarthritis Research Society International score, the levels of interleukin-1β, interleukin-6, tumor necrosis factor-α, nitric oxide, malondialdehyde, matrix metalloproteinase 3 and the expressions of nuclear factor-κB p65, Toll-like receptors 4, and myeloid differentiation factor 88 were significantly increased in the lipopolysaccharide group, while the superoxide dismutase activity and the expressions of Aggrecan and collagen type II were significantly decreased (P < 0.05). (3) To conclude, astragalus polysaccharide can improve extracellular matrix degradation, inflammatory response, and oxidative stress levels, and alleviate joint injury due to knee osteoarthritis by inhibiting the activation of Toll-like receptor 4/nuclear factor κB p65 pathway. 

Key words: astragalus polysaccharides, knee osteoarthritis, inflammation, Toll-like receptor 4, nuclear factor κB p65, rat

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