AIM:To explore the influence of ethyl(2,4,6-trimethylbenzoyl)phenylphosphinate(TPOL)on cell apoptosis and its potential mechanism.METHODS:HEK293T cells sensitive to TPOL were treated with different concentrations of T...AIM:To explore the influence of ethyl(2,4,6-trimethylbenzoyl)phenylphosphinate(TPOL)on cell apoptosis and its potential mechanism.METHODS:HEK293T cells sensitive to TPOL were treated with different concentrations of TPOL with or without exposure to light radiation,before treatment with various inhibitors,N-acetyl-Lcysteine(NAC),pifithrin-αand Z-DVED-FMK.Cell viability was measured by CCK-8 assay.Annexin V/propidium iodide staining was used to count the number of apoptotic cells.DCFH-DA staining was used to detect reactive oxygen species(ROS)levels,and JC-1 staining was used to assess mitochondrial membrane potential by flow cytometry.The expression of apoptosis-related proteins and cell cycle-regulated molecules was measured by Western blot.RESULTS:TPOL enhanced the apoptosis of HEK293T cells in a dose-dependent manner(P<0.05),with a decrease in Bcl-2 and increases in Bax and cytochrome C(Cyto C),followed by up-regulation of activated caspase-9 and caspase-3,and the cleavage of PARP(P<0.05).The TPOL-enhanced cleavage of caspase-3 and PARP was rescued by Z-DVED-FMK(P<0.01).TPOL also led to a rapid increase in ROS,a reduction in mitochondrial membrane potential,and the release of Cyto C(P<0.01),all of which could be reversed by the ROS scavenger NAC.Moreover,the TPOL-caused alterations in p21,p27,Rb,and CDK2 were also recovered by the p53 inhibitor pifithrin-α(P<0.05).The TPOL-induced changes in Bax,Bcl-2,cleaved caspase-9,activated caspase-3,and cleaved PARP were subsequently rescued by pretreatment with pifithrin-α(P<0.05).CONCLUSION:TPOL can induce cellular apoptosis with ROS-mediated mitochondrial membrane damage through the activation of a ROS-dependent p53/p21/p27/Rb/Bax/Cyto C/caspase-mediated signal axis.展开更多
Spodoptera litura is the most threatening pest in lotus production,seriously affecting the lotus yield and quality.Emamectin benzoate and chlorantraniliprole,the main insecticides for controlling S.litura on vegetable...Spodoptera litura is the most threatening pest in lotus production,seriously affecting the lotus yield and quality.Emamectin benzoate and chlorantraniliprole,the main insecticides for controlling S.litura on vegetables,are widely used by farmers to control S.litura on lotus plants.To determine the application concentrations,control effects,and safety of the two insecticides in lotus fields,indoor experiments were conducted to determine the control effects of 200 g/L chlorantraniliprole(SC)and 5%emamectin benzoate(WDG),and the residues of the two insecticides in the water,lotus leaves,and lotus seeds after field application were determined by HPLC-MS/MS.The indoor experiment results showed that chlorantraniliprole and emamectin benzoate both had good control effects on S.litura,with the median lethal concentrations(LC50)of 17.700 and 1.694 mg/L,respectively.After unmanned aerial vehicle spraying of emamectin benzoate at 20 g/667m^(2),there was no residue of emamectin benzoate in the water or lotus leaves after 5 d.After spraying of chlorantraniliprole at 20 mL/667m^(2),the residual amounts in the water and lotus leaves after 9 d were 0.005 and 0.007 mg/L,respectively.No residue of the two insecticides was detected in lotus seeds(dry and fresh)2 h after spraying.Therefore,it was recommended that chlorantraniliprole and emamectin benzoate can be used to control S.litura in lotus fields during the growth period,while attention should be paid to the application interval for safety.Considering the safe harvesting of lotus seeds and leaves,it was recommended that the preharvest intervals of chlorantraniliprole and 5%emamectin benzoate should be 9 d.展开更多
基金Supported by the National Natural Science Foundation of China(No.81172824)。
文摘AIM:To explore the influence of ethyl(2,4,6-trimethylbenzoyl)phenylphosphinate(TPOL)on cell apoptosis and its potential mechanism.METHODS:HEK293T cells sensitive to TPOL were treated with different concentrations of TPOL with or without exposure to light radiation,before treatment with various inhibitors,N-acetyl-Lcysteine(NAC),pifithrin-αand Z-DVED-FMK.Cell viability was measured by CCK-8 assay.Annexin V/propidium iodide staining was used to count the number of apoptotic cells.DCFH-DA staining was used to detect reactive oxygen species(ROS)levels,and JC-1 staining was used to assess mitochondrial membrane potential by flow cytometry.The expression of apoptosis-related proteins and cell cycle-regulated molecules was measured by Western blot.RESULTS:TPOL enhanced the apoptosis of HEK293T cells in a dose-dependent manner(P<0.05),with a decrease in Bcl-2 and increases in Bax and cytochrome C(Cyto C),followed by up-regulation of activated caspase-9 and caspase-3,and the cleavage of PARP(P<0.05).The TPOL-enhanced cleavage of caspase-3 and PARP was rescued by Z-DVED-FMK(P<0.01).TPOL also led to a rapid increase in ROS,a reduction in mitochondrial membrane potential,and the release of Cyto C(P<0.01),all of which could be reversed by the ROS scavenger NAC.Moreover,the TPOL-caused alterations in p21,p27,Rb,and CDK2 were also recovered by the p53 inhibitor pifithrin-α(P<0.05).The TPOL-induced changes in Bax,Bcl-2,cleaved caspase-9,activated caspase-3,and cleaved PARP were subsequently rescued by pretreatment with pifithrin-α(P<0.05).CONCLUSION:TPOL can induce cellular apoptosis with ROS-mediated mitochondrial membrane damage through the activation of a ROS-dependent p53/p21/p27/Rb/Bax/Cyto C/caspase-mediated signal axis.
基金Supported by Agricultural Science and Technology Innovation Fund Project of Hunan Province(2023CX98)Hunan Vegetable Industry Technology System(HARS-04)Fund for Innovation and Entrepreneurship of Technological Enterprises in Yueyang City(2023081700020)。
文摘Spodoptera litura is the most threatening pest in lotus production,seriously affecting the lotus yield and quality.Emamectin benzoate and chlorantraniliprole,the main insecticides for controlling S.litura on vegetables,are widely used by farmers to control S.litura on lotus plants.To determine the application concentrations,control effects,and safety of the two insecticides in lotus fields,indoor experiments were conducted to determine the control effects of 200 g/L chlorantraniliprole(SC)and 5%emamectin benzoate(WDG),and the residues of the two insecticides in the water,lotus leaves,and lotus seeds after field application were determined by HPLC-MS/MS.The indoor experiment results showed that chlorantraniliprole and emamectin benzoate both had good control effects on S.litura,with the median lethal concentrations(LC50)of 17.700 and 1.694 mg/L,respectively.After unmanned aerial vehicle spraying of emamectin benzoate at 20 g/667m^(2),there was no residue of emamectin benzoate in the water or lotus leaves after 5 d.After spraying of chlorantraniliprole at 20 mL/667m^(2),the residual amounts in the water and lotus leaves after 9 d were 0.005 and 0.007 mg/L,respectively.No residue of the two insecticides was detected in lotus seeds(dry and fresh)2 h after spraying.Therefore,it was recommended that chlorantraniliprole and emamectin benzoate can be used to control S.litura in lotus fields during the growth period,while attention should be paid to the application interval for safety.Considering the safe harvesting of lotus seeds and leaves,it was recommended that the preharvest intervals of chlorantraniliprole and 5%emamectin benzoate should be 9 d.
