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目的 探讨骨关节炎中长链非编码RNA转移相关肺腺癌转录物1(MALAT1)/微小RNA-15b-5p(miR-15b-5p)调控Wnt/β-catenin通路在脂多糖(LPS)诱导软骨损伤中的分子机制。方法 以5 mg/L LPS处理ATDC5细胞形成骨关节炎细胞损伤模型，RT-qPCR检测细胞中MALAT1和miR-15b-5p的表达。MTT、流式细胞术、茜素红染色和ELISA检测MALAT1和miR-15b-5p对LPS诱导的软骨细胞损伤的影响。双荧光素酶报告基因实验检验MALAT1和miR-15b-5p的调控关系。Western blot检测相关蛋白的表达情况。结果 在LPS诱导的ATDC5细胞中，MALAT1表达降低(P<0.05)。与Control组比较，LPS组细胞活性降低，凋亡率升高，肿瘤坏死因子-α、白细胞介素-6、白细胞介素-1β水平升高，钙化结节增多，MMP13、ADAMTS5蛋白表达水平升高，CollagenⅡ、Aggrecan蛋白表达水平降低，Wnt1和β-catenin蛋白表达水平升高(P<0.05)。MALAT1过表达可消减LPS对软骨细胞活性、凋亡、炎症反应、成骨分化、细胞外基质降解和Wnt/β-catenin通路的影响(P<0.05);而miR-15b-5p过表达增强了LPS对软骨细胞的作用(P<0.05)。结论 MALAT1在LPS诱导的软骨细胞中低表达，其通过靶向miR-15b-5p抑制Wnt/β-catenin通路减轻LPS诱导的软骨细胞损伤。
Abstract：Objective To explore the molecular mechanism of long non-coding RNA metastasis associated lung adenocarcinoma transcript 1(MALAT1)/microRNA-15b-5p(miR-15b-5p) regulating the Wnt/β-catenin pathway in lipopolysaccharide(LPS)-induced chondrocyte injury in osteoarthritis. Methods ATDC5 cells were treated with 5 mg/L LPS to establish the osteoarthritis cell injury model, and the expression levels of MALAT1 and miR-15b-5p in the cells were detected by RT-qPCR. The MTT, flow cytometry, Alizarin red staining, and ELISA were used to assess the effects of MALAT1 and miR-15b-5p on LPS-induced chondrocyte injury. The dual-luciferase reporter gene assay was used to examine the regulatory relationship between MALAT1 and miR-15b-5p. Western blot assay was used to evaluate the expression of relevant proteins. Results In LPS-induced ATDC5 cells, MALAT1 expression decreased(P<0.05). Compared to the control group, the LPS group exhibited reduced cell activity, an increased apoptosis rate, elevated levels of tumor necrosis factor-α, interleukin-6, and interleukin-1β, a higher number of calcified nodules, increased expression levels of extracellular matrix degradation-related proteins MMP13 and ADAMTS5, decreased expression levels of Collagen Ⅱ and Aggrecan, and increased protein expression levels of Wnt1 and β-catenin(P<0.05). Overexpression of MALAT1 could mitigate the effects of LPS on chondrocyte activity, apoptosis, inflammatory response, osteogenic differentiation, extracellular matrix degradation, and the Wnt/β-catenin pathway(P<0.05). Additionally, the overexpression of miR-15b-5p enhanced the impact of LPS on chondrocytes(P<0.05). Conclusion MALAT1 is lowly expressed in LPS-induced chondrocytes, and it alleviates LPS-induced chondrocyte injury by targeting miR-15b-5p to inhibit the Wnt/β-catenin pathway.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.07.010
中图分类号:R684.3
引用信息:
[1]赵志,刘梦鲲,查日飞等.LncRNA MALAT1/miR-15b-5p调控Wnt/β-catenin通路对脂多糖诱导软骨细胞损伤的影响[J].安徽医科大学学报,2025,60(07):1231-1240.DOI:10.19405/j.cnki.issn1000-1492.2025.07.010.
基金信息:
安徽省教育厅自然科学研究重点项目(编号：KJ2021A0807); 蚌埠医科大学研究生科研创新计划项目(编号：Byycxz21087)~~