摘要
目的研究氟化钠(sodium fluoride,NaF)是否通过Fas介导的死亡受体通路导致心肌细胞凋亡,探讨氟中毒致心肌损伤可能的作用机制。方法用0.24、0.48、0.96 mmol·L^(-1) NaF处理H9c2心肌细胞,以制备NaF染毒细胞模型,以不加NaF的组作为对照组。通过CCK-8法检测细胞活力变化,用TUNEL法观察心肌细胞形态和凋亡情况,用Real-time PCR和Western blot方法检测心肌细胞中凋亡相关因子的mRNA和蛋白表达水平。结果NaF可抑制H9c2细胞增殖,改变细胞形态,诱导细胞凋亡,且与染毒剂量及染毒时间有关。与对照组相比,NaF处理H9c2细胞24、48、72 h后,随NaF浓度的增加,Fas、caspase-1、caspase-8、caspase-3 mRNA表达水平明显增加。染毒48 h后,随NaF浓度的增加,H9c2细胞Fas、caspase-1、caspase-8、cleaved caspase-3蛋白表结论NaF可能通过Fas介导的死亡受体通路影响与凋亡相关的基因与蛋白的表达,导致心肌细胞凋亡,进而造成心肌细胞损伤。
Aim To investigate whether sodium fluoride(NaF)could induce cardiomyocyte apoptosis by Fas mediated death receptor pathway,and to explore the possible mechanism of myocardium damage caused by chronic fluorosis.Methods H9c2 cardiomyocytes cultured in vitro were treated with 0.24,0.48 and 0.96 mmol·L^(-1) NaF to establish the model of fluoride exposure.CCK-8 method was used to detect changes in cell viability.TUNEL method was used to observe cardiomyocyte morphology and apoptosis.Real-time PCR and Western blot were employed to detect the mRNA and protein expressions of Fas,caspase-1,caspase-8,caspase-3 in cardiomyocytes.Results After treatment of H9c2 cardiomyocyte with NaF,it was found that NaF significantly inhibited H9c2 cardiomyocyte proliferation,changed cell morphology,and induced cardiomyocyte apoptosis.And these changes were related to the dose and time of NaF exposure.In comparison with control group,mRNA expressions of Fas,caspase-1,caspase-8,caspase-3 of H9c2 cardiomyocytes were up-regulated gradually after treatment with different concentrations of NaF for 24,48,and 72 hours.After 48 hours,with the increase of NaF concentration,the protein expressions of Fas,caspase-1,caspase-8 and cleaved caspase-3 in H9c2 cardiomyocytes were significantly up-regulated,which were significantly higher than those in control group.Conclusion NaF may induce cardiomyocyte apoptosis by Fas-mediated death receptor pathway through affecting the expression of genes and proteins related to apoptosis.
作者
徐薇
曹丽婷
石瑞丽
齐瑞芳
郝肖琼
马宝慧
XU Wei;CAO Li-ting;SHI Rui-li;QI Rui-fang;HAO Xiao-qiong;MA Bao-hui(Department of Physiology, Baotou Medical College,Baotou Inner Mongolia 014040, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第12期1715-1720,共6页
Chinese Pharmacological Bulletin
基金
内蒙古自治区高等学校科学研究项目(No NJZY21047,NJZY214)
内蒙古自然科学基金项目(No 2015MS0851)。
