磷酸三(1,3-二氯-2-丙基)酯对人肾小管上皮细胞HKC的毒性效应及凋亡机制研究  

Toxic effects and apoptotic mechanism of tris(1,3-dichloro-2-propyl)phosphate on human renal tubular epithelial cells

作　　者：丰一兴[1] 杨奕[1] 李明[1] 施嘉琛[1] 邵兵[1] FENG Yi-xing;YANG Yi;LI Ming;SHI Jia-chen;SHAO Bing(Beijing Center for Disease Prevention and Control,Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning,Beijing 100013,China)

机构地区：[1]北京市疾病预防控制中心,食物中毒诊断溯源技术北京市重点实验室,北京100013

出　　处：《毒理学杂志》2023年第2期93-98,107,共7页Journal of Toxicology

基　　金：高层次公共卫生技术人才建设项目资助(学科骨干-02-15)。

摘　　要：目的 探讨磷酸三(1,3-二氯-2-丙基)酯[tris(1,3-dichloro-2-propyl) phosphate, TDCPP]对人肾小管上皮细胞(human renal tubular epithelial cells, HKC)的凋亡作用,并基于线粒体凋亡途径探讨其效应机制。方法 选取HKC作为受试细胞,经TDCPP(0、25、50、75和100μmol/L)染毒48 h后,利用CCK-8方法检测HKC的细胞存活率,通过高内涵成像系统观察细胞形态,采用基于细胞成像的多参数分析技术,检测TDCPP对线粒体膜电位(mitochondrial membrane potential, MMP)、细胞凋亡及线粒体凋亡途径相关蛋白表达量的影响。结果 随着TDCPP暴露浓度的增加,HKC存活率明显降低(F=54.671;P<0.05)。TDCPP浓度高于75μmol/L时,细胞形态发生明显改变,MMP明显降低(F=24.923;P<0.05),细胞凋亡率增加(F=29.303;P<0.01),线粒体凋亡途径相关蛋白,包括Bcl-2相关X蛋白(B_(ax))、B淋巴细胞瘤-2(Bcl-2)、细胞色素c和活化的半胱氨酸蛋白酶(cleaved Caspase-3)的表达量以及B_(ax)/Bcl-2的比值均明显增加(F_(B_(ax))=21.152,PB_(ax)<0.01;FBcl-2=118.788,PBcl-2<0.05;F细胞色素c=40.576,P细胞色素c<0.01;F_(caspase-3)=70.067,P_(caspase-3)<0.01;F_(B_(ax))/Bcl-2=12.192,P_(B_(ax)/Bcl-2)<0.01;)。结论 TDCPP可引起HKC线粒体受损,MMP下降,细胞凋亡率增加,线粒体凋亡途径在TDCPP引起的肾细胞凋亡中起到一定的调控作用。Objective To investigate the effect of tris(1,3-dichloro-2-propyl)phosphate(TDCPP)on the apoptosis of human renal tubular epithelial cells(HKC)through the mitochondria-dependent pathway of apoptosis.Methods HKC was selected as a cell model,after being treated with different concentrations of TDCPP(0,25,50,75 and 100μmol/L)for 48 h,cell viability of HKC was detected by CCK8 assay.Cell morphology was observed through high content imaging system.Multi-parameter analysis technology based on cell imaging was used to examine the effects of TDCPP on mitochondrial membrane potential,apoptosis as well as the protein expression levels of mitochondria-mediated apoptosis pathway.Results Result showed that cell viability was decreased significantly with the increased concentration of TDCPP.In addition,at the concentration up to 75μmol/L,alteration in cell morphology,decrease in mitochondrial membrane potential,increase in cell apoptosis,increased ration of Bax/Bcl-2 and up-regulated protein expression(Bax,Bcl-2,Cytochrome c and cleaved Caspase-3)of mitochondrial apoptosis pathway were observed.Conclusion TDCPP treatment can cause damage of mitochondria which decreases mitochondrial membrane potential and might eventually lead to the mitochondria-dependent apoptosis of HKC.

关 键 词：磷酸三(1 3-二氯-2-丙基)酯 人肾细胞 细胞凋亡 线粒体凋亡途径 

分 类 号：R99[医药卫生—毒理学] R114[医药卫生—药学]