地塞米松通过抑制p53蛋白降解诱导成骨细胞凋亡  

Dexamethasone induces apoptosis in osteoblast cells by inhibiting p53 protein degradation

作　　者：山枫 朱杰[1] 文军 梁磊 印春花 刘尧[1] 方建峰[1] 甄允方[1] SHAN Feng;ZHU Jie;WEN Jun;LIANG Lei;YIN Chunhua;LIU Yao;FANG Jianfeng;ZHEN Yunfang(Department of Pediatric Orthopedics,Children's Hospital of Soochow University,Suzhou,Jiangsu 215000,China)

机构地区：[1]苏州大学附属儿童医院儿童骨科,江苏苏州215000

出　　处：《安徽医药》2023年第5期888-893,共6页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(82172520)。

摘　　要：目的观察地塞米松对成骨细胞凋亡的影响并探讨其作用机制。方法分别采CellTiter-Glo®荧光细胞活力检测试剂盒和Annexin V-FITC/PI双染凋亡检测试剂盒测定地塞米松作用后MC3T3-E1细胞的增殖和凋亡。蛋白质印迹法(Western blotting)检测地塞米松作用后MC3T3-E1细胞和hFOB1.19细胞胱天蛋白酶3(caspase-3)/裂解caspase-3和PARP/裂解PARP的表达。用Western blotting和PCR分析地塞米松作用后MC3T3-E1细胞中p53和检控点激酶2(Chk2)的表达。探讨p53敲除和Chk2敲除在地塞米松诱导MC3T3-E1细胞凋亡中的作用。结果地塞米松能显著抑制MC3T3-E1细胞的生长并诱导其凋亡。地塞米松能通过促进p53的表达诱导成骨细胞凋亡。地塞米松对P53蛋白表达的调节作用是通过上调Chk2来实现的,Chk2与P53相互作用,抑制P53蛋白的降解。P53基因敲除可通过降低裂解caspase-3和裂解PARP的表达来抑制地塞米松诱导的MC3T3-E1细胞凋亡,经Dex处理的MC3T3-E1和hFOB1.19细胞中Ceaved PARP(0.38±0.02比0.20±0.01、0.29±0.02比0.20±0.02)和Ceaved caspase-3蛋白表达增加(均P<0.001)。结论地塞米松增加了Chk2蛋白的表达,稳定了p53蛋白的表达,进而促进成骨细胞的凋亡。Objective To observe the effect of dexamethasone on the apoptosis of osteoblasts and explore its mechanism.Methods The cell proliferation and apoptosis of MC3T3-E1 cells after Dex treatment were determined by the CellTiter-Glo Luminescent Cell Viability Assay kit and Annexin V-FITC/PI Double Staining Apoptosis Detection Kit,respectively.The expressions of caspase-3/Cleaved-caspase-3 and PARP/Cleaved-PARP in MC3T3-E1 cells and hFOB1.19 cells after Dex treatment were determined by western blotting.The expressions of p53 and Chk2 in MC3T3-E1 cells after Dex treatment were analyzed by western blotting and qRT-PCR.The effects of p53 knockdown and Chk2 knockdown on Dex-induced apoptosis of MC3T3-E1 cells were explored.Results Dex can remarkably inhibit cell growth and induce apoptosis of MC3T3-E1 cells.Dex induced osteoblast apoptosis by promoting p53 expression.The regulatory effect of Dex on p53 expression is mediated by its upregulation of Chk2,which interacted with p53 and inhibited p53protein degradation.Knockdown of p53 alleviated Dex-induced MC3T3-E1 cell apoptosis by decreasing the expressions of cleaved-caspase 3 and Cleaved PARP(0.38±0.02 vs.0.20±0.01,0.29±0.02 vs.0.20±0.02).Conclusion Dex increased Chk2 protein expression,which stabilizes the protein expression of p53,in turn promoting osteoblasts apoptosis.

关 键 词：地塞米松 细胞凋亡 成骨细胞 P53 成骨 

分 类 号：R965[医药卫生—药理学]