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[甘草附子汤对大鼠骨关节炎及关节软骨蛋白质组学的影响]

[Effect of Gancao Fuzi Decoction on osteoarthritis and proteomics of articular cartilage in rats].

作者信息

Huang Si-Min, Wu Yu-Meng, Zhang Li, Xie Hui-Hui, Yang Shuo, Xu Peng, Liu Bo

机构信息

School of Pharmacy, Jiangxi University of Traditional Chinese Medicine Nanchang 330004, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2021 Feb;46(3):661-669. doi: 10.19540/j.cnki.cjcmm.20201210.401.

Abstract

The rat osteoarthritis model was replicated by injection of sodium iodoacetate into the knee joint cavity, and the effects of Gancao Fuzi Decoction on rat osteoarthritis and the proteome of articular cartilage were investigated. Sixty SD rats weighing 230-250 g were randomly divided into normal group, model group, glucosamine sulfate group, and Gancao Fuzi Decoction high, medium and low dose groups. Osteoarthritis model was induced by intra-articular injection of sodium iodoacetate(3 mg on each leg) in all groups except the normal group. After modeling, each administration group was given intragastric administration for 1 month. During the administration period, joint pain test and joint width measurement were performed every week to observe the autonomous behavior of rats. Enzyme linked immunosorbent assay(ELISA) method was used to detect the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), matrix metalloproteinase-3(MMP-3), and matrix metalloproteinase tissue inhibitor(TIMP-1) in rat joint lavage fluid. Hematoxylin-eosin(HE) staining was used to observe bone and joint morphology. Nano-LC-LTQ-Orbitrap system was used to detect arti-cular cartilage proteins. The results showed that, compared with the model group, Gancao Fuzi Decoction could significantly improve joint pain and joint swelling in osteoarthritis rats, significantly reduce the contents of TNF-α, IL-1β and MMP-3 in the joint cavity la-vage fluid, increase the content of TIMP-1, and relieve inflammatory diseases such as enlarged joint space, rough cartilage edge, different thickness of cartilage layer, and disordered arrangement of chondrocytes. After comparing the proteins between the groups, 273 differential proteins were screened out. KEGG analysis found that the above differential proteins involved 43 signaling pathways such as systemic lupus erythematosus, among which 11 signaling pathways were related to osteoarthritis. The above results indicated that Gancao Fuzi Decoction had a preventive effect on osteoarthritis, and its mechanism of action may be accomplished by regulating the protein expression of osteoarthritis-related signal pathways.

摘要

通过向膝关节腔注射碘乙酸钠复制大鼠骨关节炎模型,研究甘草附子汤对大鼠骨关节炎及关节软骨蛋白质组的影响。将60只体重230 - 250 g的SD大鼠随机分为正常组、模型组、硫酸氨基葡萄糖组以及甘草附子汤高、中、低剂量组。除正常组外,其余各组均通过关节腔内注射碘乙酸钠(每只腿3 mg)诱导骨关节炎模型。造模后,各给药组进行1个月的灌胃给药。给药期间,每周进行关节疼痛测试和关节宽度测量,观察大鼠的自主行为。采用酶联免疫吸附测定(ELISA)法检测大鼠关节灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、基质金属蛋白酶-3(MMP-3)和基质金属蛋白酶组织抑制剂(TIMP-1)的含量。采用苏木精-伊红(HE)染色观察骨关节形态。使用纳升液相色谱-线性离子阱-轨道阱系统检测关节软骨蛋白。结果表明,与模型组相比,甘草附子汤可显著改善骨关节炎大鼠的关节疼痛和关节肿胀,显著降低关节腔灌洗液中TNF-α、IL-1β和MMP-3的含量,增加TIMP-1的含量,并缓解关节间隙增宽、软骨边缘粗糙、软骨层厚度不一以及软骨细胞排列紊乱等炎症病变。组间蛋白质比较后,筛选出273种差异蛋白质。KEGG分析发现,上述差异蛋白质涉及系统性红斑狼疮等43条信号通路,其中11条信号通路与骨关节炎相关。上述结果表明,甘草附子汤对骨关节炎具有预防作用,其作用机制可能是通过调节骨关节炎相关信号通路的蛋白质表达来实现的。

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