氟化钠通过氧化应激和内质网应激途径诱导PC12细胞凋亡  

Sodium fluoride induces apoptosis of PC12 via oxidative stress and endoplasmic reticulum stress pathways

作　　者：戴姗姗[1,2] 李锦沂 杨克宇 李征阳 刘飞[1,2] DAI Shanshan;LI Jinyi;YANG Keyu;LI Zhengyang;LIU Fei(Key Laboratory of Shaanxi Province for Craniofacial Precision Medical Research,College of Stomatology,Xi’an Jiaotong University,Xi’an 710004,China;Department of Pediatric Dentistry,College of Stomatology,Xi’an Jiaotong University;Department of Pediatric Dentistry,Xi’an Children’s Hospital)

机构地区：[1]西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室,西安710004 [2]西安交通大学口腔医院儿童口腔科 [3]西安市儿童医院儿童口腔科

出　　处：《山西医科大学学报》2023年第8期1107-1112,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81801310);陕西省青年科技新星项目(2023KJXX-034);中央高校基本科研业务费资助项目(xzy012021068)。

摘　　要：目的 探讨氟化钠对PC12细胞凋亡的影响及机制。方法 采用0,0.5,1,2,4 mmol/L浓度梯度的氟化钠培养基分别培养PC12细胞6,12,24,48 h。通过CCK-8法检测细胞活力,筛选构建氟中毒PC12细胞模型所需采用的氟化钠浓度和作用时间。将0,2,4 mmol/L氟化钠溶液培养24 h的PC12细胞进行AnnexinⅤ-FIFC/PI双染流式细胞术验证细胞凋亡状况;生化方法检测细胞内超氧化物歧化酶(SOD)以及丙二醛(MDA)的表达水平;RT-PCR及Western blot检测内质网应激标志物葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)的mRNA及蛋白表达水平。结果 0,2,4 mmol/L氟化钠分别作用PC12细胞24 h,细胞活力与氟化钠成剂量依赖性降低,差异具有统计学意义(P<0.05)。显微镜下观察细胞形态和流式细胞术验证了细胞凋亡率与氟化钠浓度呈剂量依赖性增加,差异具有统计学意义(P<0.05)。与0 mmol/L氟化钠相比,2,4 mmol/L氟化钠作用下细胞内SOD水平降低,而MDA的水平增加(均P<0.01);GRP78的mRNA及蛋白表达水平增加(P<0.05)。结论 氟化钠可通过细胞氧化应激及内质网应激介导细胞凋亡通路,诱导PC12细胞凋亡。Objective To explore the effect of sodium fluoride(NaF)on the apoptosis of PC12 cells and the underlying mechanism.Methods The PC12 cells were cultured in vitro with 0,0.5,1,2,4 mmol/L NaF for 6,12,24,48 h.The PC12 cell vitality was detected by CCK-8 to screen the concentration and the incubation time of NaF for the following experiments.After PC12 cells were treated with 0,2,4 mmol/L NaF for 24 h,the cell apoptosis was detected with AnnexinⅤ-FIFC/PI apoptosis kit.Superoxide dismutase(SOD)and malondialdehyde(MDA)levels were detected with relevant kits,respectively.Meanwhile,the relative mRNA and protein levels of glucose regulated protein 78(GRP78)were examined by RT-PCR and Western blotting.Results The vitality of PC12 cells was dose-dependently decreased after cultured with 0,2,4 mmol/L NaF for 24 h(P<0.05).The apoptosis rate of PC12 cells was increased dose-dependently by microscope and flow cytometry(P<0.05).Compared with 0 mmol/L NaF group,the content of SOD in PC12 cells was decreased while the content of MDA was increased in 2,4 mmol/L NaF group(P<0.01),and the relative mRNA and protein levels of GRP 78 were increased significantly(P<0.05).Conclusion NaF could induce the apoptosis of PC12 cells through oxidative stress and endoplasmic reticulum stress signaling pathway.

关 键 词：氟化钠 PC12细胞 氧化应激 内质网应激 细胞凋亡 

分 类 号：R363[医药卫生—病理学]