下调FGFR3表达加重对小鼠关节软骨表层细胞的损伤

doi:10.16352/j.issn.1001-6325.2024.11.1530

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (11): 1530-1537.doi: 10.16352/j.issn.1001-6325.2024.11.1530

下调FGFR3表达加重对小鼠关节软骨表层细胞的损伤

关云博^1,2, 李超¹, 徐成¹, 孙笑非¹, 白雪东¹, 何勍¹, 王祖强^1,2*

1.解放军总医院第六医学中心骨科,北京 100048;
2.安徽医科大学第五临床医学院,安徽合肥 230032

收稿日期:2024-08-26 修回日期:2024-09-19 出版日期:2024-11-05 发布日期:2024-10-31
通讯作者: ^*wangzqtmmu@hotmail.com
基金资助:
国家自然科学基金(82102594)

Down-regulation of FGFR3 expression aggravates the damage of articular chondrocyte superficial zone cells in mice

GUAN Yunbo^1,2, LI Chao¹, XU Cheng¹, SUN Xiaofei¹, BAI Xuedong¹, HE Qing¹, WANG Zuqiang^1,2*

1. Department of Orthopedics, the Sixth Medical Center, General Hospital of Chinese PLA, Beijing 100048;
2. The Fifth Clinical Medical College of Anhui Medical University, Hefei 230032, China

Received:2024-08-26 Revised:2024-09-19 Online:2024-11-05 Published:2024-10-31
Contact: ^*wangzqtmmu@hotmail.com

摘要/Abstract

摘要： 目的探究成纤维细胞生长因子受体3(FGFR3)对关节软骨表层细胞(SPZCs)损伤的影响。方法将小鼠随机分为2组:假手术组(sham组)、手术诱导的内侧半月板不稳定模型组(DMM组)。于术后4周和8周使用臧红固绿染色观察关节软骨组织学形态;免疫组化染色检测细胞凋亡情况及FGFR3蛋白表达情况。提取小鼠原代SPZCs,随机分为对照组和Fgfr3敲降处理组,用RT-qPCR与Western blot检测软骨细胞外基质合成、降解及软骨细胞肥大化相关基因和蛋白质表达。结果与sham组比较,DMM组术后小鼠膝关节软骨出现SPZCs率先损伤,免疫组化提示软骨细胞凋亡增加、基质金属蛋白酶13(MMP-13)及FGFR3表达降低(P＜0.05)。原代SPZCs通过转染小干扰RNA(siRNA)敲降Fgfr3,RT-qPCR结果显示软骨细胞外基质合成相关基因蛋白聚糖(Acan)和Ⅱ型胶原蛋白(Col2) mRNA表达降低,细胞外基质降解相关基因Mmp13、血小板反应蛋白解整合素金属肽酶5(Adamts5) mRNA表达升高(P＜0.05)。Western blot与RT-qPCR结果一致,显示MMP-13蛋白表达水平显著升高,而collagen Ⅱ和aggrecan蛋白表达出现显著下降(P＜0.05)。结论 Fgfr3敲降可诱导小鼠原代SPZCs出现损伤,导致早期骨关节炎(OA)发生。

关键词: 成纤维细胞生长因子受体3, 软骨表层细胞, 骨关节炎

Abstract: Objective To investigate the effect of fibroblast growth factor receptor 3 (FGFR3) on articular cartilage superficial zone cells (SPZCs). Methods C57 mice were randomly divided into two groups: a sham operated group (sham group) and a group of surgically induced unstable medial meniscus model group (DMM group). The histological morphology of articular cartilage was microscopied by Safranin O/Fast Green-stained in 4 weeks and 8 weeks after surgery. Apoptosis and FGFR3 protein expression were detected by immunohistochemical staining microscopy. Primary SPZCs were separated and randomly divided into control group and Fgfr3 knockdown treatment group. The genes and protein expression related to chondrocyte extra cellular matrix synthesis, degradation and chondrocytehypertrophy were detected by RT-qPCR and Western blot. Results Compared with the sham group, the keen cartilage of mice in DMM group showed a pioneer damage of SPZCs after surgery; Immunohistochemistry results showed an increase in chondrocyte apoptosis and a decrease in expression of MMP-13 and FGFR3(P＜0.05). Primary SPZCs were transfected with small interfering RNA (siRNA) to knockdown Fgfr3; RT-qPCR results showed that the mRNA expression of genes related to the synthesis of cartilage extracellular matrix aggrecan and Col2 was reduced; And the mRNA expression of extracellular matrix degradation-related genes Mmp13 and Adamts5 was increased. The mRNA expression of chondrocyte hypertrophy-related genes Col10 and Mmp13 was increased. Western blot and RT-qPCR results were consistent and the expression l of MMP13 protein was significantly increased, while the expression of collagen Ⅱ and aggrecan protein was significantly decreased (P＜0.05). Conclusions Knockdown of Fgfr3 induces damage to primary SPZCs in mice resulting in early osteoarthritis (OA) development.

Key words: fibroblast growth factor receptor 3, chondrocyte superficial zone cell, osteoarthritis

中图分类号:

R684.3

关云博, 李超, 徐成, 孙笑非, 白雪东, 何勍, 王祖强. 下调FGFR3表达加重对小鼠关节软骨表层细胞的损伤[J]. 基础医学与临床, 2024, 44(11): 1530-1537.

GUAN Yunbo, LI Chao, XU Cheng, SUN Xiaofei, BAI Xuedong, HE Qing, WANG Zuqiang. Down-regulation of FGFR3 expression aggravates the damage of articular chondrocyte superficial zone cells in mice[J]. Basic & Clinical Medicine, 2024, 44(11): 1530-1537.

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下调FGFR3表达加重对小鼠关节软骨表层细胞的损伤

Down-regulation of FGFR3 expression aggravates the damage of articular chondrocyte superficial zone cells in mice

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