牛膝总皂苷对骨关节炎模型兔软骨修复及低氧诱导因子1信号通路的影响

Influence of Achyranthes bidentata saponins on the repair of cartilage and hypoxia-inducible factor 1 signaling pathway in a rabbit model of osteoarthritis

背景:以往研究证实,牛膝能促进实验兔骨关节炎模型关节软骨修复,牛膝总皂苷含药血清体外能促进软骨细胞增殖。目的:观察牛膝总皂苷对实验兔骨关节炎模型软骨修复及低氧诱导因子1信号通路的影响。方法:新西兰大白兔32只由广东省实验动物中心提供,实验方案经广州中医药大学动物实验伦理委员会批准(批准号:2018023)。将32只实验兔进行骨关节炎模型造模,将造模成功的30只随机分为牛膝总皂苷组、硫酸氨基葡萄糖组、空白对照组,每组10只,分别予牛膝总皂苷、硫酸氨基葡萄糖、蒸馏水灌胃30 d,进行软骨组织苏木精-伊红染色观察、病理学改良Mankin’s评分及软骨细胞电镜观察,用qRT-PCR检测软骨组织低氧诱导因子1α、血管内皮生长因子、Ⅱ型胶原mRNA表达。结果与结论:①灌胃30 d后牛膝总皂苷组与硫酸氨基葡萄糖组、空白对照组比较,苏木精-伊红染色观察、病理学改良Mankin’s评分明显改善(P <0.05),电镜观察软骨细胞活力增加;②牛膝总皂苷组与硫酸氨基葡萄糖组、空白对照组比较,qRT-PCR检测软骨组织低氧诱导因子1α、血管内皮生长因子mRNA表达下调及Ⅱ型胶原mRNA表达上调(P <0.05);③牛膝总皂苷能有效提高细胞活力、促进软骨细胞增殖,下调低氧诱导因子1α、血管内皮生长因子mRNA表达及上调Ⅱ型胶原mRNA表达,说明低氧诱导因子1信号通路在牛膝总皂苷对实验兔骨关节炎模型软骨修复过程中发挥了较大作用。

BACKGROUND: Preliminary study has shown that Achyranthes bidentata can promote articular cartilage repair in rabbit model of osteoarthritis and Achyranthes bidentata saponins-containing serum can contribute to chondrocyte proliferation in vitro.OBJECTIVE: To observe the influence of Achyranthes bidentata saponins on the repair of cartilage and the hypoxia-inducible factor 1 signaling pathway in a rabbit model of osteoarthritis.METHODS: Thirty-two New Zealand white rabbits were provided by the Guangdong Provincial Laboratory Animal Center, and the study was approved by the Ethics Committee of Guangdong University of Chinese Medicine, approval number: 2018023. The rabbits were used for establishing the osteoarthritis model, and 30 rabbit models were randomized into three groups(n=10/group), followed by given Achyranthes bidentata saponins, glucosamine sulfate and distilled water(blank control group) via gavage, respectively, for 30 days. The cartilage was observed by hematoxylin-eosin staining, assessed by modified Mankin’s score and electron microscope. The mRNA expression levels of hypoxia-inducible factor 1α, vascular endothelial growth factor, and collagen type II in cartilage tissue were detected by qRT-PCR.RESULTS AND CONCLUSION:(1) The hematoxylin-eosin staining results and modified Mankin’s score were significantly improved in the Ayranthes bidentata saponins group compared with the glucosamine sulfate and blank control groups 30 days after treatment(P < 0.05), and electron microscope showed that the chondrocyte activity was increased.(2) Compared with the glucosamine sulfate and blank control groups,the mRNA expression levels of hypoxia-inducible factor 1α and vascular endothelial growth factor were significantly down-regulated, and the mRNA expression level of collagen type II was significantly up-regulated in the Ayranthes bidentata saponins group(P < 0.05).(3) To conclude, Ayranthes bidentata saponins can effectively improve the cell vitality, promote chondrocyte proliferation, down-regulate the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor mRNA and up-regulate the expression of collagen type mRNA, suggesting that hypoxia-inducible factor 1 signaling pathway plays a critical role in cartilage repair of rabbit model of osteoarthritis by Ayranthes bidentata saponins.