赵轶男,孙 强,毕 龙,陈小超,程建岗.miR-130a-3p调控SOX4对骨关节炎软骨细胞增殖、分化和炎症因子释放的影响[J].,2019,19(22):4201-4207 |
miR-130a-3p调控SOX4对骨关节炎软骨细胞增殖、分化和炎症因子释放的影响 |
miR-130a-3p Regulates the Proliferation, Differentiation and Release of Inflammatory Factors of Osteoarthritis Chondrocytes through miR-130a-3p |
投稿时间:2019-04-26 修订日期:2019-05-28 |
DOI:10.13241/j.cnki.pmb.2019.22.001 |
中文关键词: miR-130a-3p 骨关节炎 SOX4 增殖 分化 炎症因子 |
英文关键词: miR-130a-3p Osteoarthritis SOX4 Proliferation Differentiation Inflammatory cytokine |
基金项目:国家自然科学基金项目(2016YFC1100304) |
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中文摘要: |
摘要 目的:探讨miR-130a-3p对骨关节炎(osteoarthritis, OA)软骨细胞增殖、分化和炎症因子释放的影响及作用机制。方法:收集我院住院的40例半月板损伤患者和40例OA患者,采用RT-PCR检测半月板损伤患者和OA患者膝关节软骨组织中miR-130a-3p的表达。在OA软骨细胞中分别转染miR-NC、miR-130a-3p mimics、miR-130a-3p inhibitors、siRNA-SOX4(si-SOX4)或过表达SOX4的慢病毒载体(LV- SOX4),采用CCK8法检测细胞增殖情况;RT-PCR和western blot检测细胞分化相关分子BMP2和BMP4;RT-PCR检测炎症因子IFN-γ和TNF-α的表达。结果:半月板损伤患者软骨组织和软骨细胞中miR-130a-3p的表达明显高于OA患者(P均<0.05),而SOX4的表达水平明显低于OA患者(P<0.05)。OA患者膝关节软骨组织中miR-130a-3p和SOX4的表达呈显著负相关(P<0.05)。miR-130a-3p mimics能够明显促进OA软骨细胞增殖(P<0.05)、增加分化相关分子BMP2和BMP4的表达(P均<0.05)以及抑制炎症因子IFN-γ和TNF-α的表达(P均<0.05)。miR-130a-3p inhibitors能够明显抑制OA软骨细胞增殖(P<0.05)、分化相关分子BMP2和BMP4的表达(P均<0.05)以及促进炎症因子IFN-γ和TNF-α的表达(P均<0.05)。OA软骨细胞在转染miR-130a-3p mimics后,SOX4 mRNA和蛋白的表达水平明显降低(P均<0.05),在转染miR-130a-3p inhibitors后,SOX4 mRNA和蛋白的表达水平明显升高(P均<0.05)。双荧光素酶结果显示miR-130a-3p能够靶向结合SOX4。在OA软骨细胞中,采用siRNA低表达SOX4后,明显促进细胞增殖和分化以及抑制炎症因子的释放。共转染miR-130a-3p mimics和LV- SOX4能够逆转过表达miR-130a-3p对OA软骨细胞增殖、分化和炎症因子释放的影响(P均>0.05),而共转染miR-130a-3p inhibitors和LV- SOX4能够进一步加强低表达miR-130a-3p对OA软骨细胞增殖、分化和炎症因子的影响(P均<0.05)。结论:miR-130a-3p在OA中明显低表达,并能够通过靶向抑制SOX4促进OA软骨细胞增殖、分化和炎症因子释放。 |
英文摘要: |
ABSTRACT Objective: To investigate the effects of miR-130a-3p on osteoarthritis (OA) chondrocyte proliferation, differentiation and inflammation, and the underlying mechanisms. Methods: 40 patients with OA and 40 patients with meniscus injury in our hospital were selected. miR-130a-3p expression in OA and meniscus injury patients were analyzed by real-time polymerase chain reaction (RT-PCR). After transfecting OA chondrocytes with miR-NC, miR-130a-3p mimics, miR-130a-3p inhibitors, si-SOX4 or LV- SOX4, cell proliferation was detected by the cell counting kit-8 (CCK8) assay, BMP2 and BMP4 were detected by RT-PCR and western blotting, and the expression of IFN-γ and TNF-α were detected by RT-PCR. Results: The expression level of miR-130a-3p in patients with meniscus injury and normal chondrocytes was higher than that in OA patients (P<0.05). However, the expression level of SOX4 was lower than that in OA patients (P<0.05). The expression level of miR-130a-3p was significantly negatively related with the expression level of SOX4 in OA cartilage tissues(P<0.05). After OA chondrocytes transfected with miR-130a-3p mimics, OA chondrocyte proliferation was significantly increased(P<0.05), the expression of BMP2 and BMP4 were significantly increased(P<0.05) and the expression of IFN-γ and TNF-α were significantly decreased(P<0.05). After OA chondrocytes transfected with miR-130a-3p inhibitors, OA chondrocyte proliferation was significantly decreased(P<0.05), the expression of BMP2 and BMP4 were significantly decreased(P<0.05) and the expression of IFN-γ and TNF-α were significantly increased (P<0.05). After transfection with miR-130a-3p mimics, the expression levels of SOX4 mRNA and protein were significantly decreased in OA chondrocytes (P <0.05), while the expression levels of SOX4 mRNA and protein were significantly increased after transfection with mir-130a-3p inhibitors (P<0.05). Double luciferase results showed that miR-130a-3p could bind SOX4 in a targeted manner(P<0.05). Low expression of SOX4 by siRNA in OA chondrocytes could significantly promote cell proliferation, differentiation, and inhibit the release of inflammatory cytokine. After co-transfection with miR-130a-3p mimics and LV-SOX4, the effects of overexpression of miR-130a-3p on cell proliferation and differentiation and the release of inflammatory factors in OA chondrocytes could be reversed (P<0.05). However, after co-transfection with miR-130a-3p inhibitors and LV-SOX4, the effects of low expression of miR-130a-3p on cell proliferation and differentiation and inflammatory factors in OA chondrocytes could be further enhanced (all P<0.05). Conclusion: miR-130a-3p was significantly low expressed in OA and could regulate the proliferation, differentiation and inflammatory cytokine release in OA chondrocytes through targeting SOX4. |
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