Agkistrodon ameliorates pain response and prevents cartilage degradation in monosodium iodoacetate-induced osteoarthritic rats by inhibiting chondrocyte hypertrophy and apoptosis.
Agkistrodon ameliorates pain response and prevents cartilage degradation in monosodium iodoacetate-induced osteoarthritic rats by inhibiting chondrocyte hypertrophy and apoptosis.
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Agkistrodon 通过抑制软骨细胞肥大和凋亡来改善碘乙酸钠诱导的骨关节炎大鼠的疼痛反应并防止软骨退化。
DOI：
10.1016/j.jep.2018.12.004
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发表时间：
2019-03
影响因子：
5.4
通讯作者：
Efferth Thomas
中科院分区：
文献类型：
--
作者：
Wang Caiwei;Yan Li;Yan Bo;Zhou Li;Sun Wan;Yu Lingying;Liu Fucun;Du Wenxi;Yu Guangping;Hu Zhengyan;Yuan Qiang;Xiao Luwei;Li Hongwen;Tong Peijian;Zhang Jida;Shan Letian;Efferth Thomas
民族药理学相关性骨关节炎（OA）以关节疼痛和软骨退化为特征，是全世界最常见的关节疾病，但目前尚无令人满意的治疗方法。尖吻蝽乙醇提取物(EAA)作为中药被广泛用于治疗关节疼痛和炎症性疾病，但迄今为止尚未见其对骨关节炎的疗效报道。在此，我们在体内测定了EAA对疼痛行为和软骨退化的影响，并在体外阐明了其与软骨细胞肥大和凋亡相关的靶基因和蛋白。 材料与方法通过向大鼠关节内注射（1.5mg）碘乙酸钠（MIA）建立体内OA模型，并每周进行关节内给药治疗 EAA的剂量范围为0.3至0.9g/kg，持续四个星期。治疗前后测试疼痛行为参数、热缩回潜伏期（TWL）和机械缩回阈值（MWT）。然后对关节软骨进行组织病理学、免疫组织化学和TUNEL分析，然后进行Mankin评分。在体外，通过细胞活力、免疫荧光、实时PCR和Western blot检测EAA对软骨细胞的影响。应用UPLC-MS测定EAA的化学成分。结果动物数据显示，EEA不仅可以减轻OA大鼠的疼痛超敏性，而且可以以剂量依赖的方式通过提高软骨细胞存活率和抑制软骨细胞凋亡来阻止软骨退变。此外，EAA 显着恢复 OA 大鼠软骨中 II 型胶原蛋白 (Col2) 和基质金属蛋白酶 13 (MMP13) 的异常表达。细胞数据显示，EAA 显着增加软骨细胞抵抗 OA 样损伤的细胞活力，并恢复受损软骨细胞中 Col2 和 MMP13 的异常表达。分子数据显示，EAA可显着恢复病理状态下软骨细胞中Col2、Col10、MMP2和MMP13的异常mRNA表达以及MMP13、PARP（总和裂解）的异常蛋白表达。 UPLC-MS 分析显示 EAA 的已知主要成分，包括氨基酸（甘氨酸、L-天冬氨酸、L-谷氨酸和 L-羟脯氨酸）、核苷（尿苷）、嘌呤（黄嘌呤和次黄嘌呤）和嘧啶（尿嘧啶）。结论我们的数据表明 EAA 具有镇痛和镇痛作用。 通过抑制软骨细胞肥大和凋亡以及恢复软骨细胞中合成代谢和分解代谢的分子表达来对 OA 发挥软骨保护作用。它为治疗骨关节炎提供了一种有前途的中药候选新药。
Ethnopharmacological relevanceOsteoarthritis (OA), characterized by joint pain and cartilage degradation, is the most common form of joint disease worldwide but with no satisfactory therapy available. The ethanol extract ofAgkistrodon acutus(EAA) has been widely used as a traditional Chinese medicine (TCM) for the treatment of arthralgia and inflammatory diseases, but there is no report regarding its efficacy on OA to date. Here, we determined the effects of EAA on the pain behavior and cartilage degradation in vivo and clarified its target genes and proteins associated with chondrocyte hypertrophy and apoptosis in vitro.Materials and methodsIn vivoOA model was established by intra-articular injection (1.5 mg) of monosodium iodoacetate (MIA) into rats and weekly treated by intra-articular administration of EAA at a dose range from 0.3 to 0.9 g/kg for four weeks. The pain behavior parameters, thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were tested before and after the treatment. Then histopathologic, immunohistochemical and TUNEL analyses of the articular cartilage were conducted, followed by Mankin′s scoring.In vitro, the effects of EAA on chondrocytes were evaluated via assays of cell viability, immunofluorescence, real time PCR, and Western blot. UPLC-MS was applied to determine the chemical composition of EAA.ResultsThe animal data showed that EEA not only attenuated the pain hypersensitivity but also blocked the cartilage degeneration by improving chondrocyte survival and suppressing chondrocyte apoptosis at a dose-dependent manner in OA rats. Furthermore, EAA remarkably restored the abnormal expression of collagen type II (Col2) and matrix metalloproteinase-13 (MMP13) in cartilage of OA rats. The cellular data showed that EAA significantly increased the cell viability of chondrocytes against OA-like damage and restored the abnormal expressions of Col2 and MMP13 in damaged chondrocytes. The molecular data showed that EAA significantly restored the abnormal mRNA expressions of Col2, Col10, MMP2 and MMP13 as well as the abnormal protein expressions of MMP13, PARP (total and cleaved) in chondrocytes under pathological condition. UPLC-MS analysis showed the known main components of EAA, including amino acides (glycine,L-aspartic acid,L-glutamic acid, and L-hydroxyproline), nucleoside (uridine), purines (xanthine and hypoxanthine), and pyrimidine (uracil).ConclusionsOur data demonstrate that EAA exerts antinociceptive and chondroprotective effects on OA through suppressing chondrocyte hypertrophy and apoptosis with restoration of the molecular expressions of anabolism and catabolism in chondrocytes. It provides a promising TCM candidate of novel agent for OA therapy.
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影响因子：
7.4
作者：
Ivanavicius, Stefan P.;Ball, Adrian D.;Read, Simon J.
通讯作者：
Read, Simon J.
滑膜巨噬细胞和巨噬细胞产生的细胞因子在驱动聚集蛋白聚糖酶、基质金属蛋白酶和骨关节炎中其他破坏性和炎症反应中的作用。
影响因子：
4.9
作者：
Bondeson J;Wainwright SD;Lauder S;Amos N;Hughes CE
通讯作者：
Hughes CE
影响因子：
--
作者：
Moreau M;Pelletier JP;Lussier B;d'Anjou MA;Blond L;Pelletier JM;del Castillo JR;Troncy E
通讯作者：
Troncy E
影响因子：
--
作者：
Hegedus, Bela;Viharos, Laszlo;Galfi, Marta
通讯作者：
Galfi, Marta
影响因子：
2.3
作者：
Neogi, Tuhina;Zhang, Yuqing
通讯作者：
Zhang, Yuqing