“Sirt1-自噬”在低氧诱导小鼠成牙骨质细胞凋亡中的作用  

Role of“Sirt1-autophagy”on hypoxia-induced apoptosis in mice cementoblasts

作　　者：谭玺 罗虹 武红彦 黄兰[1] Tan Xi;Luo Hong;Wu Hongyan;Huang Lan(Stomatological Hospital of Chongqing Medical University,Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences,Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education)

机构地区：[1]重庆医科大学附属口腔医学院、口腔疾病与生物医学重庆市重点实验室、重庆市高校市级口腔生物医学工程重点实验室,重庆401147

出　　处：《重庆医科大学学报》2023年第1期24-29,共6页Journal of Chongqing Medical University

基　　金：国家自然科学基金青年科学基金资助项目(编号:81300914);2016年重庆高校创新团队建设计划资助项目(编号:CXTDG201602006);重庆市高校市级口腔生物医学工程重点实验室资助项目。

摘　　要：目的:初步研究“Sirt1-自噬”在低氧诱导小鼠成牙骨质细胞OCCM-30凋亡中的作用。方法:体外培养小鼠成牙骨质细胞,随机分为对照组、低氧组、低氧+Sirt1激活剂白藜芦醇组、低氧+自噬抑制剂3-MA组、低氧+白藜芦醇+Sirt1抑制剂ex527组、低氧+白藜芦醇+3-MA组,MDC染色观察自噬溶酶体变化;LC3Ⅱ免疫荧光检测自噬体变化;透射电镜观察自噬体;Western blot检测Sirt1、LC3Ⅱ、p62等蛋白表达;流式细胞仪检测细胞凋亡情况;不同组间结果数据采用单因素方差分析,组间采用LSD-t分析。结果:低氧组较对照组自噬溶酶体含量(P=0.000)及自噬体数量(P=0.000)明显增加,LC3Ⅱ蛋白表达明显增加(P=0.001),p62蛋白表达明显减少(P=0.000),细胞凋亡率增加(P=0.000);低氧+白藜芦醇组较低氧组自噬溶酶体(P=0.019)及自噬体数量(P=0.000)、LC3Ⅱ蛋白表达(P=0.049)、Sirt1蛋白表达(P=0.000)明显增加,p62蛋白表达明显减少(P=0.000),细胞凋亡率减少(P=0.021);低氧+白藜芦醇+ex527组较低氧+白藜芦醇组自噬溶酶体(P=0.000)及自噬体数量(P=0.000)、LC3Ⅱ蛋白表达(P=0.000)、Sirt1蛋白表达(P=0.000)明显减少,p62蛋白表达明显增加(P=0.000),细胞凋亡率增加(P=0.000);低氧+白藜芦醇+3-MA组较低氧+白藜芦醇组自噬溶酶体(P=0.000)及自噬体数量(P=0.000)、LC3Ⅱ蛋白表达明显减少(P=0.000),p62蛋白表达明显增加(P=0.000),细胞凋亡率增加(P=0.000),Sirt1蛋白表达减少(P=0.146),差异无统计学意义。结论:自噬相关分子可能是低氧环境下Sirt1的下游作用分子,自噬在低氧诱导小鼠成牙骨质细胞(OCCM-30)凋亡中起保护作用。Objective:To investigate the role of“Sirt1-autophagy”on the hypoxia-induced apoptosis in OCCM-30 cementoblasts of mouse.Methods:OCCM-30 cells were cultured in vitro and were randomly divided into the control group,the hypoxia group,the hypoxia+resveratrol group,the hypoxia+3-MA group,the hypoxia+resveratrol+ex527 group and the hypoxia+resveratrol+3-MA group.Changes of autophagy lysosomes in each group were observed by MDC staining and changes of autophagosome light chainⅡ(LC3Ⅱ)were detected by immunofluorescence assay.The expression of Sirt1,LC3Ⅱand p62 proteins was detected by Western blot.Cell apoptosis was detected by flow cytometry.Results of different groups were analyzed by one-way ANOVA and different groups were analyzed by LSD-t.Results:Compared with the control group,the content of autophagy lysosomes(P=0.000)and the number of autophagosomes(P=0.000)in the hypoxia group were significantly increased,the expression of LC3Ⅱ(P=0.001)protein was significantly increased,the expression of p62 protein was significantly decreased(P=0.000),and the apoptosis rate was increased(P=0.000).Compared with the hypoxia group,the number of autophagy lysosomes(P=0.019)and autophagosomes(P=0.000),and the expression of LC3Ⅱprotein(P=0.000)and Sirt1 protein(P=0.049,P=0.000)in the hypoxia+resveratrol group were significantly increased,the expression of p62 protein(P=0.000)was significantly decreased,and the apoptosis rate was decreased(P=0.021).Compared with the hypoxia+resveratrol group,the number of autophagy lysosomes(P=0.000)and autophagosomes(P=0.000),and the expression of LC3Ⅱ(P=0.000)and Sirt1(P=0.000)in the hypoxia+resveratrol+ex527 group were significantly decreased,the expression of p62 protein(P=0.000)was significantly increased,and the apoptosis rate was increased(P=0.000).Compared with the hypoxia+resveratrol group,the number of autophagy lysosomes(P=0.000)and autophagosomes(P=0.000),and the expression of LC3Ⅱ(P=0.000)protein expression in the hypoxia+resveratrol+3-MA group were significantly decre

关 键 词：成牙骨质细胞 SIRT1 自噬 低氧 凋亡 

分 类 号：R783.3[医药卫生—口腔医学]