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山东大学学报 (医学版) ›› 2022, Vol. 60 ›› Issue (10): 1-8.doi: 10.6040/j.issn.1671-7554.0.2022.0439

• 基础医学 •    下一篇

促甲状腺激素通过抗炎蛋白CTRP3促进软骨细胞分化

张薇薇1,2,华芳3,梁超帅2,褚苗苗2,孙嘉忆4,Frank Zaucke5,辛玮2,4   

  1. 1.山东大学齐鲁医学院, 山东 济南 250012;2.山东第一医科大学附属省立医院中心实验室, 山东 济南 250021;3.临沂市人民医院输血科, 山东 临沂 276003;4.山东第一医科大学临床与基础医学院, 山东 济南 250021;5.歌德大学法兰克福大学医院 骨科, 法兰克福 德国 60388
  • 发布日期:2022-09-30
  • 通讯作者: 辛玮. E-mail:vivienxin@126.com
  • 基金资助:
    国家自然科学基金(81970427,81471007)

Thyroid stimulating hormone promotes chondrocyte differentiation via anti-inflammatory protein CTRP3

ZHANG Weiwei1,2, HUA Fang3, LIANG Chaoshuai2, CHU Miaomiao2, SUN Jiayi4, FRANK Zaucke5, XIN Wei2,4   

  1. 1. Cheeloo College of Medicine, Shandong University, Jinan 250012, Shandong, China;
    2. Central Laboratory, Shandong Provincial Hospital, Shandong First Medical University, Jinan 250021, Shandong, China;
    3. Department of Blood Transfusion, Linyi Peoples Hospital of Shandong Province, Linyi 276003, Shandong, China;
    4. School of Clinical and Basic Medicine, Shandong First Medical University, Jinan 250021, Shandong, China;
    5. Department of Orthopaedics University Hospital Frankfurt, Goethe University 60388, Frankfurt, Germany
  • Published:2022-09-30

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摘要： 目的 探讨促甲状腺激素(TSH)对软骨细胞(PMCs)分化的作用及相关机制。 方法 利用qRT-PCR、Western blotting、免疫荧光检测PMCs细胞外基质Ⅱ型胶原(Col Ⅱ)、Ⅹ型胶原(Col Ⅹ)以及基质金属蛋白酶3和13(MMP3,MMP13)的表达变化。采用qRT-PCR、ELISA检测TSH刺激PMCs后炎症因子白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNFα)的表达水平。应用高通量RNA-seq技术筛选TSH调控PMCs分化的差异表达基因,通过在原代PMCs中过表达CTRP3,探索TSH调控PMCs分化的机制。 结果 qRT-PCR、Western blotting结果显示,TSH刺激下PMCs中MMP3(P=0.016,P<0.001)和MMP13(P均<0.001)的表达显著增加。qRT-PCR、ELISA结果显示,IL-1β(P=0.007,P=0.002)、IL-6(P均<0.001)、TNFα(P=0.006,P<0.001)的表达上调。通过RNA-seq筛选出差异表达基因CTRP3。CTRP3过表达后PMCs中Col Ⅱ表达增高(F=76.062,F=77.085,F=190.115)、Col Ⅹ表达降低(F=33.494,F=38.424,F=43.351);同时与单纯TSH刺激相比,MMP3(F=88.607,F=214.145,F=135.60)、MMP13(F=116.561,F=138.674,F=86.865)表达均显著降低(P均<0.001),IL-1β、IL-6、TNFα的表达则均降低(P均<0.001)。 结论 CTRP3对TSH诱导的PMCs分化具有保护作用,有望成为亚临床甲状腺功能减退症(SCH)患者关节损伤的潜在治疗靶点。

关键词: 骨关节炎, 促甲状腺激素, 软骨细胞分化, C1q/肿瘤坏死因子相关蛋白3, 炎症因子

Abstract: Objective To investigate the role of thyroid stimulating hormone(TSH)in the differentiation of primary mouse chondrocytes(PMCs)and the underlying mechanism. Methods The expressions of chondrocyte differentiation markers, including extracellular matrix collagen type Ⅱ(Col Ⅱ)and collagen type Ⅹ(Col Ⅹ), and matrix metalloproteinase 3(MMP3)and matrix metalloproteinase 13(MMP13)were detected on both mRNA and protein levels with qRT-PCR, Western blotting and immunofluorescent staining. The changes of interleukin-1β(IL-1β), interleukin-6(IL-6)and tumor necrosis factor-α(TNFα)were detected in TSH stimulated group and control group with qRT-PCR and ELISA. The differentially expressed genes of PMCs treated with TSH were screened with high-throughput RNA-Seq. PMCs were transfected with C1q/tumor necrosis factor-related protein 3(CTRP3)overexpression plasmid o explore the mechanism of TSH regulating PMCs differentiation. Results TSH upregulated the expressions of MMP3(P=0.016, P<0.001), MMP13(P<0.001), IL-1β(P=0.007, P=0.002), IL-6(P<0.001)and TNFα(P=0.006, P<0.001). The differentially expressed gene CTRP3 was screened out with RNA-seq. After CTRP3 overexpression, the expression of Col Ⅱ in PMCs was increased(F=76.062, F=77.085, F=190.115, P<0.001), while the expression of Col Ⅹ was decreased(F=33.494, F=38.424, F=43.351). Compared with simple TSH stimulation, after CTRP3 overexpression, the expressions of MMP3 (F=88.607, F=214.145, F=135.60), MMP13(F=116.561, F=138.674, F=86.865), IL-1β, IL-6 and TNFα were significantly decreased(P<0.001). Conclusion CTRP3 has a protective effect on TSH-induced differentiation of PMCs, and it may become a promising anti-inflammatory therapeutic target for joint injury in patients with subclinical hypothyroidism(SCH).

Key words: Osteoarthritis, Thyroid stimulating hormone, Primary mouse chondrocytes differentiation, C1q/tumor necrosis factor-related protein 3, Inflammatory cytokines

中图分类号: 

  • R446
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