百草枯诱导AML12细胞凋亡过程中线粒体活性氧的作用

doi:10.3969/j.issn.1004-616x.2016.01.001

Abstract

Abstract: OBJECTIVE: To investigate the role of mitochondrial reactive oxygen species (mtROS) on the apoptosis of a mouse liver cell line-AML12 cells after their exposure to paraquat. METHODS: AML12 cells were treated with paraquat at concentrations of 0,25,50,100,200 and 300 μmol/L for 24 h,and cell viability was measured by MTT assay. Apoptosis was assessed by flow cytometry using Annexin V-FITC apoptosis detection kit. DCFH-DA fluorescent probe and MitoSOX were used to determine the level of ROS in whole cell and in mitochondria by flow cytometry,respectively. Mitochondrial membrane potential was evaluated by confocal microscope using JC-1 probe. RESULTS: Paraquat from 25 to 300 μmol/L induced dose-dependent decrease of AML12 cells viability. Apoptosis was significantly induced by paraquat at 300 μmol/L for 24 h. Treatment with 25-100 μmol/L paraquat raised the level of mitochondrial reactive oxygen species. However,200 and 300 μmol/L paraquat decreased the level of mtROS. The increase of ROS level was closely related to the decrease of AML12 cells viability (r=-0.90,P<0.05),and the ROS level was positively correlated with the proportion of the late apoptotic cells (r=0.96,P<0.01). Additionally,300 μmol/L paraquat markedly reduced mitochondrial membrane potential. CONCLUSION: Paraquat could induce apoptosis of AML12 cells via the induction of whole cell ROS which suggest that mtROS may not be directly involved in paraquat-induced liver injury.

Key words: paraquat, reactive oxygen species, mitochondrial reactive oxygen species, mitochondrial membrane potential, apoptosis

CLC Number:

R994.6

KONG Deqin, LIU Jiangzheng, YU Weihua, ZHANG Tao, LONG Zi, WANG Xin, ZHANG Xiaodi, BAI Hua, HAI Chunxu. Involvement of mitochondrial reactive oxygen species on paraquat-induced apoptosis in AML12 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(1): 1-7.

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Involvement of mitochondrial reactive oxygen species on paraquat-induced apoptosis in AML12 cells

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