纳米二氧化硅通过PI3K/AKt信号通路影响肺泡巨噬细胞凋亡的研究  

Nano-silicon Dioxide Affects Apoptosis of Alveolar Macrophages via PI3K/AKt Signaling Pathway

作　　者：李宁 陈祥娃[1] 霍婷婷 董发勤[3] 邓建军[1,4] LI Ning;CHEN Xiang-wa;HUO Ting-ting;DONG Fa-qin;DENG Jian-jun(Department of Laboratory Medicine,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Laboratory Medicine,Sichuan Provincial Hospital for Women and Children,Chengdu 610041,China;School of Environment and Resource,Southwest University of Science and Technology,Mianyang 621010,China;Department of Laboratory Medicine,Sichuan Mianyang 404 Hospital,Mianyang 621000,China)

机构地区：[1]西南医科大学附属医院医学检验部,泸州646000 [2]四川省妇幼保健院检验科,成都610041 [3]西南科技大学固体废物处理与资源化教育部重点实验室,绵阳621010 [4]四川绵阳四〇四医院检验科,绵阳621000

出　　处：《四川大学学报（医学版）》2020年第4期488-493,共6页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.41472046,No.41602033);四川省科技计划项目(No.2016JY0045)资助。

摘　　要：目的探讨磷脂酰肌醇激酶-3/蛋白激酶B(phosphatidyl inositol 3-kinase/protein kinase B,PI3K/AKt)信号通路对纳米二氧化硅(nano silica,NS)粉尘诱导肺泡巨噬细胞(alveolar macrophages,AM)凋亡发生的影响。方法通过不同质量浓度的NS粉尘染毒AM细胞后,CCK-8法检测细胞存活率;荧光显微镜观察细胞形态;流式细胞术检测PI3K抑制剂LY294002预处理前后细胞的凋亡率与线粒体膜电位;Western blot法检测PI3K抑制剂LY294002预处理前后细胞凋亡相关蛋白Bax、Bcl-2以及磷酸化(p)-PI3K、p-AKt的表达水平。结果NS可降低AM细胞的存活率,部分细胞出现凋亡形态学改变;LY294002预处理AM后,线粒体膜电位水平的下降程度增加,并下调Bcl-2、p-PI3K、p-AKt蛋白的表达,同时上调Bax蛋白的表达,增加细胞凋亡率。结论PI3K/AKt信号通路可能参与了NS诱导AM细胞的凋亡过程。Objective To investigate the effect of phosphatidyl inositol 3-kinase/protein kinase B(PI3 K/AKt)signaling pathway on the apoptosis of alveolar macrophages(AM)induced by nano-silica(NS)dust.Methods After exposure to different concentrations of NS suspension,CCK-8 assay was used to detect the AM viability;the cellular morphology of apoptotic AM was observed under fluorescence microscopy;the apoptosis rate and mitochondrial transmembrane potential of cells were detected by flow cytometry before and after pretreatment with phosphatidyl inositol 3-kinase(PI3 K)inhibitor LY294002;Western blot was used to detect the expression of apoptosis-related proteins Bax,Bcl-2,p-PI3 K and p-AKt.Resluts The survival rate of AM was decreased in a time-dose relationship after NS exposure.With LY294002 pretreatment,the mitochondrial transmembrane potential level and the expressions of p-PI3 K,p-AKt and Bcl-2 were decreased,the expression of Bax and the apoptosis rate were increased.Conclusion Our data suggested that the activation of PI3 K/AKt signaling pathway played an important role in NS-induced apoptosis in alveolar macrophages.

关 键 词：纳米二氧化硅 肺泡巨噬细胞 PI3K/AKT 细胞凋亡 

分 类 号：R114[医药卫生—卫生毒理学]