流体静压力通过Ca^(2+)/CaMKⅡ/MAPK信号通路调控髁突软骨细胞增殖与凋亡  

Hydrostatic Pressure Regulates Proliferation and Apoptosis of Condylar Chondrocytes through Ca^(2+) /CaMKⅡ/MAPK Signaling Pathway

作　　者：梁秋娟[1] 热依拉·艾克兰木 肖朋 LIANG Qiujuan;Reyila·AIKELANMU;XIAO Peng(Department of Stomatology,Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,China;Department of Stomatology,Seventh Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,China.)

机构地区：[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

出　　处：《口腔医学研究》2024年第4期315-320,共6页Journal of Oral Science Research

基　　金：新疆维吾尔自治区自然科学基金(编号:2022D01C315)。

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Ca^(2+)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 kinaseⅡ,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Ca^(2+)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Ca^(2+)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Ca^(2+)水平,其作用机制为Ca^(2+)通过CaMKⅡ激活丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPK)信号通路。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 Ca^(2+)levels.Western blot was used to detect the expression of CaMKⅡ,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 Ca^(2+)levels increased significantly,the expression of CaMKⅡprotein 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 Ca^(2+)level decreases,the expression of CaMKⅡprotein 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 Ca^(2+)levels.The mechanism of action may be that Ca^(2+)activates the MAPK signaling pathway through CaMKⅡ.

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 

分 类 号：R782.6[医药卫生—口腔医学]