BACKGROUND Osteoarthritis (OA) is a frequent and incurable joint disease, inducing significant pain and seriously threatening to human health. It has been reported that microRNAs (miRNAs) play crucial roles on cancers and inflammatory diseases via cooperating with genes. However, the effect of miR-374a-3p/Wingless-type MMTV integration site family, member 5B (WNT5B) pair in OA remains to be explored. METHODS GSE105027 and GSE55457 datasets were obtained to reveal the expression of miR-374a-3p and WNT5B in OA cartilages using log-scale. The OA cell model was established by lipopolysaccharides (LPS) stimulation in CHON-001 cells and the functional role of miR-374a-3p on OA was investigated by analyzing cell proliferation, cell apoptosis and the expression of apoptosis-related proteins (Bcl-2, Bax and Bim). Through bioinformatics prediction, WNT5B, the target gene of miR-374a-3p, was predicted and the association between miR-374a-3p and WNT5B was further explored by luciferase reporter assay. Functional experiments in vitro were conducted to assess whether WNT5B was involved in the regulation of miR-374a-3p to LPS-stimulated CHON-001. Finally, the expression of JNK/ERK/MAPK pathway-related proteins was detected to explore the underlying molecular mechanism. RESULTS The data set showed that miR-374a-3p was decreased in OA cartilages and the consistent expressional pattern was observed in LPS-stimulated CHON-001 cells. Overexpression of miR-374a-3p significantly alleviated LPS-induced damage in CHON-001 cells, whereas miR-374a-3p inhibitor aggravated LPS-stimulated injury. Further experiments demonstrated that WNT5B was a target of miR-374a-3p and its expression was decreased by miR-374a-3p. WNT5B expression was increased in OA cartilages. Silencing WNT5B prevented CHON-001 cells from LPS-induced damage. Down-regulation of WNT5B strengthened the protective effect of miR-374a-3p on LPS-stimulated CHON-001 cells. Moreover, miR-374a-3p cooperated with WNT5B to affect cell behaviors of LPS-stimulated CHON-001 cells via mediating the JNK/ERK/MAPK pathway. CONCLUSION These results indicated that overexpression of miR-374a-3p protects CHON-001 cells against LPS challenge by modulating WNT5B and inhibiting the JNK/ERK/MAPK pathway.

中文翻译：

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上调的miR-374a-3p通过调节Wingless型MMTV整合位点家族成员5B减轻CHON-001细胞中脂多糖诱导的损伤。

背景技术骨关节炎（OA）是一种常见且不可治愈的关节疾病，引起严重的疼痛并严重威胁人类健康。据报道，microRNA（miRNA）通过与基因合作在癌症和炎性疾病中起关键作用。但是，在OA中，miR-374a-3p / Wingless型MMTV整合位点家族成员5B（WNT5B）对的作用尚待探索。方法获得GSE105027和GSE55457数据集，以对数刻度揭示miR-374a-3p和WNT5B在OA软骨中的表达。通过脂多糖（LPS）刺激CHON-001细胞建立OA细胞模型，并通过分析细胞增殖，细胞凋亡和凋亡相关蛋白（Bcl-2）的表达来研究miR-374a-3p在OA中的功能。 ，Bax和Bim）。通过生物信息学预测WNT5B，预测了miR-374a-3p的靶基因，并通过荧光素酶报告基因检测进一步探索了miR-374a-3p和WNT5B之间的关联。进行了体外功能实验，以评估WNT5B是否参与miR-374a-3p对LPS刺激的CHON-001的调控。最后，检测JNK / ERK / MAPK途径相关蛋白的表达，以探索其潜在的分子机制。结果数据集显示，miR-374a-3p在OA软骨中降低，并且在LPS刺激的CHON-001细胞中观察到一致的表达模式。miR-374a-3p的过表达显着减轻了LPS诱导的CHON-001细胞的损伤，而miR-374a-3p抑制剂则加剧了LPS刺激的损伤。进一步的实验表明，WNT5B是miR-374a-3p的靶标，miR-374a-3p降低了其表达。WNT5B表达在OA软骨中增加。沉默WNT5B可防止CHON-001细胞受到LPS诱导的损伤。WNT5B的下调增强了miR-374a-3p对LPS刺激的CHON-001细胞的保护作用。而且，miR-374a-3p与WNT5B协同作用，通过介导JNK / ERK / MAPK途径影响LPS刺激的CHON-001细胞的细胞行为。结论这些结果表明，miR-374a-3p的过表达通过调节WNT5B和抑制JNK / ERK / MAPK途径，保护CHON-001细胞免受LPS攻击。miR-374a-3p与WNT5B协同作用，通过介导JNK / ERK / MAPK途径影响LPS刺激的CHON-001细胞的细胞行为。结论这些结果表明，miR-374a-3p的过表达通过调节WNT5B和抑制JNK / ERK / MAPK途径，保护CHON-001细胞免受LPS攻击。miR-374a-3p与WNT5B协同作用，通过介导JNK / ERK / MAPK途径影响LPS刺激的CHON-001细胞的细胞行为。结论这些结果表明，miR-374a-3p的过表达通过调节WNT5B和抑制JNK / ERK / MAPK途径，保护CHON-001细胞免受LPS攻击。