SMSC-EXO来源lncRNA-SNHG14对OA大鼠软骨细胞损伤及炎性痛的调节作用和机制研究-东南大学学报医学版

SMSC-EXO来源lncRNA-SNHG14对OA大鼠软骨细胞损伤及炎性痛的调节作用和机制研究

单位：海南医学院第一附属医院关节外科, 海南海口 570100

关键词：长链非编码RNA SNHG14 滑膜间充质干细胞外泌体骨性关节炎炎症疼痛大鼠

分类号：R684.3

出版年·卷·期（页码）：2023·42·第一期（22-31）

摘要：

目的：探讨滑膜间充质干细胞（SMSC）外泌体（EXO）来源的长链非编码RNA （lncRNA） SNHG14（SMSC-EXO-SNHG14）对骨关节炎（OA）中软骨细胞损伤及炎性疼痛缓解的作用和机制。方法：通过分离过表达SNHG14的SMSC的EXO，获得SMSC-EXO-SNHG14。使用SMSC-EXO-SNHG14治疗OA软骨细胞和OA大鼠。用IL-1β处理大鼠软骨细胞以建立OA的体外模型，通过CCK-8和transwell测定评估软骨细胞增殖、迁移能力。注射碘乙酸钠建立大鼠OA模型。造模6周后采集大鼠膝关节标本和背根神经节（DRG）进行组织学分析。在模型建立前和模型建立后1、2、4和6周评估机械缩爪阈值（PWT）和热缩爪潜伏期（PWL），并通过Western blotting和ELISA检测软骨降解和炎症相关标志物的表达。结果：体外SMSC-EXO-SNHG14可被软骨细胞内吞。SMSC-EXO-SNHG14处理显著减弱了IL-1β对软骨细胞增殖和迁移的抑制作用（P<0.05），IL-1β诱导的COL2A1下调和MMP13的上调（P<0.05）。SMSC-EXO-SNHG14治疗显著上调OA大鼠软骨组织中的COL2A1蛋白和下调MMP13蛋白（P<0.05）。在第2、4和6周，与未处理的OA大鼠相比，SMSC-EXO-SNHG14处理的OA大鼠的PWL显著改善（P<0.05）。此外，SMSC-EXO-SNHG14治疗显著减轻了OA大鼠DRG组织中降钙素基因相关肽（CGRP）和诱导型一氧化氮合酶（iNOS）的上调（P<0.05）。结论：SMSC-EXO-SNHG14可有效促进OA大鼠的软骨修复，以及减轻膝关节炎症和疼痛。

Objective: To investigate the effect and mechanism of long non-coding RNA(lncRNA) SNHG14(SMSC-EXO-SNHG14) derived from exosomes(EXO) of synovial mesenchymal stem cells(SMSC) on chondrocyte injury and inflammatory pain relief in osteoarthritis(OA). Methods: SMSC-EXO-SNHG14 was obtained by isolating EXO of SNHG14-overexpressing SMSC cells. OA chondrocytes and OA rats were treated with SMSC-EXO-SNHG14. Rat chondrocytes were treated with IL-1β to establish an in vitro model of OA, Chondrocyte proliferation and migration were evaluated by CCK-8 and transwell assays. Rat OA model was established by injecting sodium iodoacetate. The rat knee joint specimens and dorsal root ganglion(DRG) were collected for histological analysis 6 weeks after modeling. For pain assessment, paw withdrawal threshold(PWT) and paw withdrawal latency(PWL) were assessed before model establishment and 1, 2, 4, and 6 weeks after model establishment. The expression of markers related to cartilage degradation and inflammation was detected by Western blotting and ELISA. Results: SMSC-EXO-SNHG14 was endocytosed by chondrocytes in vitro. SMSC-EXO-SNHG14 treatment significantly attenuated the inhibitory effect of IL-1β on chondrocyte proliferation and migration(P<0.05). SMSC-EXO-SNHG14 treatment significantly attenuated IL-1β-induced down-regulation of COL2A1 and up-regulation of MMP13(P<0.05). In animal studies, SMSC-EXO-SNHG14 treatment significantly up-regulated the expression of COL2A1 and down-regulated the expression of MMP13 in cartilage tissue of OA rats(P<0.05). In 2, 4 and 6 weeks, the PWL values of SMSC-EXO-SNHG14-treated OA rats were significantly improved compared with untreated OA animals(P<0.05). In addition, SMSC-EXO-SNHG14 treatment significantly attenuated the up-regulation of CGRP and iNOS in DRG tissues of OA rats(P<0.05). Conclusion: SMSC-EXO-SNHG14 effectively promotes cartilage repair in OA rats, and reduces knee joint inflammation and pain.

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