上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (8): 861-.doi: 10.3969/j.issn.1674-8115.2019.08.009

• 论著·基础研究 • 上一篇    下一篇

胃泌素对白介素 1β诱导的大鼠软骨细胞分解代谢的抑制作用

李莹莹 1，罗亚平 2，傅国辉 1, 2，陈诗慧 1   

  1. 1. 上海交通大学基础医学院病理中心，细胞分化与凋亡教育部重点实验室，上海 200025；2. 上海交通大学附属第一人民医院病理中心，上海
  • 出版日期:2019-08-28 发布日期:2019-09-23
  • 通讯作者: 陈诗慧，电子信箱：chensh2015@shsmu.edu.cn。
  • 作者简介:李莹莹（1994—），女，硕士生；电子信箱： 289010797@qq.com。
  • 基金资助:
    上海市自然科学基金（18ZR1421800）

Inhibition of gastrin on catabolism of rat chondrocytes inducedinterleukin-1β

LI Ying-ying1, LUO Ya-ping2, FU Guo-hui1, 2, CHEN Shi-hui1   

  1. 1. Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Pathology Center, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Pathology Center, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200080, China
  • Online:2019-08-28 Published:2019-09-23
  • Supported by:
    Shanghai Natural Science Foundation，18ZR1421800

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摘要： 目的 ·探究胃泌素对白介素 1β（interleukin-1β，IL-1β）诱导的大鼠关节软骨细胞的保护作用及其机制。方法 ·原代培养并鉴定新生大鼠膝关节软骨细胞，MTT法测定胃泌素对细胞活力的影响。10 ng/mL的 IL-1β诱导软骨细胞 2 d后，胃泌素（ 1×10-7 mol/L）处理 3 d，实时荧光定量 PCR检测软骨细胞分解代谢基因基质金属蛋白酶 3（matrix metalloproteinase 3，MMP3）和 MMP13的表达；Western blotting检测胃泌素信号通路中关键蛋白胆囊收缩素 B受体（ cholecystokinin B receptor，CCKBR）、细胞外调节蛋白激酶（extracellular regulated protein kinases，ERK）、磷酸化 ERK（phospho-ERK，p-ERK）、P65的表达；另设正常对照组和 IL-1β诱导组。结果 · 1×10-7 mol/L浓度的胃泌素无细胞毒性作用，并能明显抑制 IL-1β诱导的软骨细胞分解代谢基因 MMP3和 MMP13的高表达。与正常对照组比较， IL-1β诱导组及胃泌素处理组 CCKBR的表达均显著升高，且胃泌素处理能显著增加 p-ERK的表达，抑制 P65的表达。结论 ·胃泌素能激活 CCKBR，诱导 ERK磷酸化，抑制 P65的活性，从而拮抗 IL-1β诱导引起的 MMP3和 MMP13 mRNA表达升高，进而可能抑制大鼠软骨细胞分解代谢活动。

关键词: 胃泌素, 软骨细胞, 分解代谢, 分子机制

Abstract:

Objective · To investigate the protective effect of gastrin on rat chondrocytes treated with interleukin-1β (IL-1β) and underlying mechanism . Methods · Chondrocytes of knee joint of newborn rats were isolated and cultured. The effect of gastrin on cell viability was determinedMTT assay. In addition, after being induced with 10 ng/mL of IL-1β for 2 d, the chondrocytes were treated with 1×10-7 mol/L of gastrin for 3 d. The s of chondrocyte catabolic genes, i.e., matrix metalloproteinase 3 (MMP3) and MMP13, and the key proteins in gastrin-related signaling pathway, i.e., cholecystokinin B receptor (CCKBR), extracellular regulated protein kinases (ERK), phospho-ERK (p-ERK) and p65 were detectedreal-time PCR and Western blotting, respectively. Results · Gastrin with the concentration of 1×10-7 mol/L had no cytotoxic effects on the viability of chondrocytes. Based on IL-1β induction, the s of MMP3 and MMP13 genes were significantly downregulatedgastrin. Meanwhile, the of CCKBR in the IL-1β induction group and gastrin treatment groups was higher than that in the normal control group. Gastrin also facilitated the phosphorylation of ERK and resulted in the inhibition of P65. Conclusion · Gastrin increases the phosphorylation level of ERKactivating its receptor CCKBR, thereby inhibiting the activity of P65 in NF-κB pathway, antagonizing the chondrocyte catabolic activity inducedIL-1β.Accordingly, gastrin may possess the potentials of cartilage matrix protection via inhibiting mRNA s of MMP3 and MMP13 and catabolism in rat chondrocytes.

Key words: gastrin, chondrocyte, catabolism, molecular mechanism

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