Fu Changlong, Qiu Zhiwei, Huang Yanfeng, Mei Yangyang, Lin Qing, Zeng Jianwei, Zhong Weihong, Ma Dezun
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.
Fujian Provincial Key Laboratory of Integrative Medicine on Geriatrics, Fuzhou, Fujian 350122, P.R. China.
Exp Ther Med. 2022 Jun 22;24(2):532. doi: 10.3892/etm.2022.11459. eCollection 2022 Aug.
polysaccharides (ABPS) is an active ingredient of the flowering plant that has been previously reported to be effective for the treatment of osteoarthritis (OA). However, the underlying molecular mechanism remain to be fully clarified. Emerging studies have shown that the long non-coding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) is involved in the pathogenesis of OA. Therefore, the present study aimed to investigate the potential mechanism of ABPS by focusing on its effects on the regulation of chondrocyte extracellular matrix (ECM) homeostasis, with particular emphasis on lncRNA GAS5. In the present study, the modified Hulth method was used to construct OA rats, which were gavaged with 400 mg/kg ABPS for 8 weeks. Histopathological changes in cartilage and subchondral bone were evaluated by hematoxylin-eosin staining and Safranin O-fast green staining. In experiments, IL-1β-treated chondrocytes were infected with Lenti-lncRNA GAS5. Fluorescence in situ hybridization assay was performed to measure the expression of the lncRNA GAS5 in chondrocytes. Moreover, the relative expression level of lncRNA GAS5 in cartilage tissue and chondrocytes was detected using reverse transcription-quantitative PCR. Western blot analysis was used to detect protein expression levels of MMP-9, MMP-13, TIMP-1, TIMP-3 and type II collagen in cartilage tissue and chondrocytes. The results indicated that ABPS delayed the degradation of the ECM by chondrocytes in addition to reducing lncRNA GAS5 expression both and . Furthermore, silencing of lncRNA GAS5 expression in IL-1β-treated chondrocytes downregulated the protein expression of MMP-9 and MMP-13 whilst upregulating the expression of tissue inhibitor matrix metalloproteinase (TIMP)-1, TIMP-3 and type II collagen. To conclude, the present study provides evidence that ABPS can inhibit the expression of lncRNA GAS5 in chondrocytes to regulate the homeostasis of ECM, which in turn may delay the occurrence of cartilage degeneration during OA.
多糖（ABPS）是一种开花植物的活性成分，此前有报道称其对骨关节炎（OA）的治疗有效。然而，其潜在的分子机制仍有待充分阐明。新兴研究表明，长链非编码RNA（lncRNA）生长停滞特异性转录本5（GAS5）参与了OA的发病机制。因此，本研究旨在通过关注ABPS对软骨细胞外基质（ECM）稳态调节的影响，特别是lncRNA GAS5，来探讨其潜在机制。在本研究中，采用改良的Hulth方法构建OA大鼠，用400mg/kg ABPS灌胃8周。通过苏木精-伊红染色和番红O-固绿染色评估软骨和软骨下骨的组织病理学变化。在实验中，用慢病毒载体lncRNA GAS5感染白细胞介素-1β处理的软骨细胞。进行荧光原位杂交试验以检测软骨细胞中lncRNA GAS5的表达。此外，使用逆转录定量PCR检测软骨组织和软骨细胞中lncRNA GAS5的相对表达水平。采用蛋白质印迹分析检测软骨组织和软骨细胞中基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶-13（MMP-13）、基质金属蛋白酶组织抑制因子-1（TIMP-1）、基质金属蛋白酶组织抑制因子-3（TIMP-3）和II型胶原蛋白的蛋白表达水平。结果表明，ABPS除了降低软骨组织和软骨细胞中lncRNA GAS5的表达外，还延缓了软骨细胞对ECM的降解。此外，在白细胞介素-1β处理的软骨细胞中沉默lncRNA GAS5的表达可下调MMP-9和MMP-13的蛋白表达，同时上调基质金属蛋白酶组织抑制因子（TIMP）-1、TIMP-3和II型胶原蛋白的表达。总之，本研究提供了证据表明ABPS可抑制软骨细胞中lncRNA GAS5的表达以调节ECM的稳态，这反过来可能延迟OA期间软骨退变的发生。
