Hubei Province Key Laboratory of Occupational Hazard Identification and Control, Wuhan University of Science and Technology, Wuhan, 430065, China.
New Medicine Innovation and Development Institute, Department of Pharmacy, College of Medicine, Wuhan University of Science and Technology, Wuhan, 430065, China.
BMC Complement Med Ther. 2020 Apr 19;20(1):117. doi: 10.1186/s12906-020-02920-9.
Our previous in vitro study reported that the ethyl acetate fraction (EAF) of Semen sojae germinatum (SSG), the processed sprout of Chinese black soybean, possessed the potent anti-inflammatory and chondroprotective properties. The aim of the present work was to verify the in vivo antiosteoarthritic effect of EAF from SSG on a rat osteoarthritis (OA) model .
A classical rat OA model was surgically induced by anterior cruciate ligament transaction (ACLT). The OA rats were intra-articularly administered EAF from SSG for 8 weeks. The cartilage and synovial tissues were stained with hematoxylin and eosin (HE) to observe the histopathological changes. Safranin O/fast green staining was used to assess the glycosaminoglycan content in cartilage tissue sections. The expression of type II collagen and matrix metalloproteinase (MMP)-13 in cartilage was measured by immunohistochemistry. The apoptotic chondrocytes in the cartilage sections were detected using TUNEL assay. The concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-ɑ in synovial fluid were determined using ELISA.
Intra-articular administration of EAF from SSG well retained the structure and superficial layer of cartilage tissues, ameliorated cartilage lesion and the degradation of cartilage matrix, including proteoglycan and type II collagen, induced by ACLT operation. The ACLT-induced upregulation of MMP-13 expression in the cartilage tissues was resisted by EAF from SSG. Moreover, EAF from SSG inhibited the ACLT-induced chondrocyte apoptosis. Compared to OA model group, the inflammatory status of synovial membrane was improved, the levels of inflammatory cytokines IL-1β and TNF-ɑ in synovial fluid were decreased in rats administrated with EAF from SSG.
These data suggested that EAF from SSG displayed in vivo protective effect on OA development via preventing the degeneration of articular cartilage, inhibiting chondrocyte apoptosis and suppressing synovial inflammation.
我们之前的体外研究报告称，中国黑豆发芽后的胚轴提取物（SSG 的乙酸乙酯级分，简称 EAF）具有很强的抗炎和软骨保护特性。本工作旨在验证 SSG 的 EAF 对大鼠骨关节炎（OA）模型的体内抗 OA 作用。
通过前交叉韧带切断术（ACLT）手术诱导经典的大鼠 OA 模型。OA 大鼠接受 SSG 的 EAF 关节内给药 8 周。用苏木精和伊红（HE）染色观察软骨和滑膜组织的组织病理学变化。番红 O/快绿染色用于评估软骨组织切片中的糖胺聚糖含量。免疫组织化学法测量软骨中 II 型胶原和基质金属蛋白酶（MMP）-13 的表达。TUNEL 测定法检测软骨切片中的凋亡软骨细胞。ELISA 法测定滑液中白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-ɑ的浓度。
SSG 的 EAF 关节内给药很好地保留了软骨组织的结构和浅层，改善了软骨病变和 ACLT 手术引起的软骨基质降解，包括糖胺聚糖和 II 型胶原。SSG 的 EAF 抑制了软骨组织中 MMP-13 表达的 ACLT 诱导上调。此外，EAF 从 SSG 抑制 ACLT 诱导的软骨细胞凋亡。与 OA 模型组相比，EAF 从 SSG 处理的大鼠滑膜膜的炎症状态得到改善，滑液中炎症细胞因子 IL-1β和 TNF-ɑ的水平降低。
这些数据表明，SSG 的 EAF 通过防止关节软骨退化、抑制软骨细胞凋亡和抑制滑膜炎症，对 OA 发展具有体内保护作用。
