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

doi:10.3969/j.issn.2095-4344.1381

Abstract

Abstract:

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.

Key words: Ayranthes bidentata saponins, osteoarthritis, signaling pathway, chondrocytes, hypoxia-inducible factor 1

CLC Number:

Ma Dujun1, Peng Liping1, Yu Tian1, Cao Yafei1, Gao Kun1, Xiao Wei1, Hu Liekui2, Xu Wenming1. Influence of Achyranthes bidentata saponins on the repair of cartilage and hypoxia-inducible factor 1 signaling pathway in a rabbit model of osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(27): 4332-4337.

Figures/Tables 2

2.1 实验动物数量分析选取造模成功的新西兰大白兔30只，分为3组，实验过程无脱失，进入结果分析30只。 2.2 一般情况骨关节炎模型造模6周后按照马氏骨关节炎模型造模验证方法验证实验动物造模成功[19]，症见：造模关节僵硬、肿胀，股四头肌萎缩，关节腔见软骨颜色白硬，软骨面光泽度降低，软骨面有不同程度粗糙、糜烂、溃疡形成，关节周缘骨刺形成，造模过程中未见固定肢体出现坏死、压疮等现象，实验动物灌胃过程中未出现死亡者。 2.3 软骨组织苏木精-伊红染色、病理学改良Mankin’s评分骨关节炎模型造模成功后苏木精-伊红染色提示基质染色变浅，软骨细胞数量明显减少，血管入侵潮线结构，改良Mankin’s评分6.78±0.53。实验兔灌胃后牛膝总皂苷组苏木精-伊红染色提示基质染色无明显变化，软骨细胞数量较空白对照组增多，软骨细胞呈卵圆形，潮线结构恢复正常，改良Mankin’s评分为3.46±0.45。硫酸氨基葡萄糖组苏木精-伊红染色提示基质染色变浅，软骨细胞数量减少，血管入侵潮线结构，改良Mankin’s评分6.74±0.57。空白对照组苏木精-伊红染色提示基质染色变浅，软骨细胞数量明显减少，血管入侵潮线结构，改良Mankin’s评分6.92±0.49。牛膝总皂苷组与硫酸氨基葡萄糖组、空白对照组改良Mankin’s评分相比有统计学意义(P < 0.05)。见图1。 2.4 软骨组织电镜观察见图2。电镜醋酸铀和柠檬酸铅双染色：牛膝总皂苷组图a显示软骨细胞呈卵圆形，细胞及胞膜完整，表面微绒毛稀疏短小，胞质可见粗面内质网，部分粗面内质网扩张成空泡状，细胞核完整，核内染色质分布均匀，胞膜外软骨囊清晰可见，图b显示纤维组织排列致密，规则；图c硫酸氨基葡萄糖组显示：软骨细胞呈卵圆形，细胞及胞膜完整，表面微绒毛稀疏至缺失，胞质可见大量空泡状粗面内质网，细胞核完整但不规则，核内异染色质分布分布于核膜之下，胞膜外软骨囊不明显，图d显示纤维组织排列致密，不规则；空白对照组图e显示软骨细胞呈卵圆形，细胞及胞膜完整，表面微绒毛稀疏短小，胞质可见粗面内质网和纤维蛋白丝，部分线粒体扩张成空泡状，细胞核完整但不规则，核内异染色质分布分布于核膜之下，胞膜外软骨囊不明显，图f显示纤维组织排列致密，不规则。 2.5 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 qRT-PCR检测结果分析表明，与硫酸氨基葡萄糖组、空白对照组相比，牛膝总皂苷组低氧诱导因子1α、血管内皮生长因子mRNA表达量显著降低(P < 0.01)，与硫酸氨基葡萄糖组相比，牛膝总皂苷组Ⅱ型胶原mRNA表达量均明显增加(P < 0.05)。硫酸氨基葡萄糖组与空白剂量组比较Ⅱ型胶原mRNA表达量均明显增加(P < 0.05)，而低氧诱导因子1α、血管内皮生长因子mRNA表达量差异无显著性意义(P > 0.05)。见图3。 "

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