口腔医学研究

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (4): 315-320.DOI: 10.13701/j.cnki.kqyxyj.2024.04.007

• 口腔生物学研究 • 上一篇    下一篇

流体静压力通过Ca2+/CaMKⅡ/MAPK信号通路调控髁突软骨细胞增殖与凋亡

梁秋娟1, 热依拉·艾克兰木1, 肖朋2*   

  1. 1.新疆医科大学第五附属医院口腔科 新疆 乌鲁木齐 830000;
    2.新疆医科大学第七附属医院口腔科 新疆 乌鲁木齐 830000
  • 收稿日期:2023-12-18 发布日期:2024-04-22
  • 通讯作者: *肖朋,E-mail:17799757839lqj@sina.com
  • 作者简介:梁秋娟(1984~ )女, 新疆人, 硕士, 副主任医师, 研究方向:颞下颌关节紊乱病病因机制研究。
  • 基金资助:
    新疆维吾尔自治区自然科学基金(编号:2022D01C315)

Hydrostatic Pressure Regulates Proliferation and Apoptosis of Condylar Chondrocytes through Ca2+/CaMKⅡ/MAPK Signaling Pathway

LIANG Qiujuan1, Reyila·AIKELANMU1, XIAO Peng2*   

  1. 1. Department of Stomatology, Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, China;
    2. Department of Stomatology, Seventh Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, China
  • Received:2023-12-18 Published:2024-04-22

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摘要： 目的: 考察30 kPa流体静压力调控髁突软骨细胞增殖及凋亡的分子机制。方法: 体外培养大鼠髁突软骨细胞,分为4组:正常压力组、30 kPa压力组、KN93对照组、30 kPa压力+KN93组。5-乙炔基-2'脱氧尿嘧啶核苷( 5-ethynyl-2'-deoxyuridine,EdU)标记法测定细胞增殖。流式细胞术法检测细胞凋亡比例。荧光探针法检测细胞内Ca2+水平。Western blot法检测细胞钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinase Ⅱ,CaMKⅡ)、p-c-Jun、c-Jun、磷酸化的细胞外信号调节激酶1/2(phosphorylated extracellular signal-regulated kinase 1/2,p-ERK1/2)、细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)、p-p38、p38蛋白表达。结果: 与正常压力组相比,30 kPa压力组细胞增殖率均降低;凋亡率均提高;细胞内Ca2+水平升高;CaMKⅡ蛋白表达上调,(p-c-Jun)/(c-Jun)、(p-ERK1/2)/(ERK1/2)、(p-p38)/(p38)的比值均增加,差异具有统计学意义(P<0.01)。与30 kPa压力组相比,30 kPa压力+KN93组细胞增殖率均提高;凋亡率均降低;细胞内Ca2+水平降低;CaMKⅡ蛋白表达下调,(p-c-Jun)/(c-Jun)、(p-ERK1/2)/(ERK1/2)、(p-p38)/(p38)的比值均降低,差异具有统计学意义(P<0.01)。结论: 30 kPa静压力持续作用髁突软骨细胞,可抑制细胞增殖,促进细胞凋亡,提高细胞内Ca2+水平,其作用机制为Ca2+通过CaMKⅡ激活丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPK)信号通路。

关键词: 软骨细胞, 静压力, 增殖, 凋亡, 丝裂原活化蛋白激酶信号通路

Abstract: Objective: To investigate the molecular mechanism of 30 kPa hydrostatic pressure regulating the proliferation and apoptosis of condylar chondrocytes. Methods: Rat condylar chondrocytes were cultured in vitro. Cells divided into four groups: normal pressure group, 30 kPa pressure group, KN93 control group, and 30 kPa pressure+KN93 group. EdU labeling method was used to measure cell proliferation. Flow cytometry was used to detect the proportion of cell apoptosis. Fluorescence probe method was used to detect intracellular Ca2+ levels. Western blot was used to detect the expression of CaMK Ⅱ, p-c-Jun, c-Jun, p-ERK1/2, ERK1/2, p-p38, and p38 proteins in cells. Results: Compared with the normal pressure group, the cell proliferation rate in the 30 kPa pressure group decreased, the cell apoptosis rate increased significantly, the intracellular Ca2+ levels increased significantly, the expression of CaMK Ⅱ protein was upregulated, and the ratios of (p-c-Jun)/(c-Jun), (p-ERK1/2)/(ERK1/2), and (p-p38)/(p38) were all increased (P<0.01). Compared with the 30 kPa pressure group, the cell proliferation rate in the 30 kPa pressure+KN93 group was increased, the cell apoptosis rate decreased, the intracellular Ca2+ level decreases, the expression of CaMK Ⅱ protein was downregulated, and the ratios of (p-c-Jun)/(c-Jun), (p-ERK1/2)/(ERK1/2), and (p-p38)/(p38) were all reduced (P<0.01). Conclusion: The sustained effect of 30 kPa hydrostatic pressure can inhibit condylar chondrocytes proliferation, promote cell apoptosis, and increase intracellular Ca2+ levels. The mechanism of action may be that Ca2+ activates the MAPK signaling pathway through CaMK Ⅱ.

Key words: chondrocytes, hydrostatic pressure, proliferation, apoptosis, MAPK signaling pathway