叔丁基对苯二酚对百草枯神经细胞毒性和氧化应激的拮抗作用

目的 研究叔丁基对苯二酚(tBHQ)预处理对百草枯(PQ)致大鼠肾上腺嗜铬细胞瘤(PC12)细胞毒性和氧化应激的拮抗作用.方法 将PC12细胞分为溶剂对照组及100、300 μmol PQ处理组.用终浓度为40 μmol/LtBHQ预处理PC12细胞4 h再分别用终浓度0、100、300 μmol/L PQ处理细胞24或48 h,用噻唑蓝(MTT)法检测细胞毒性,用Annexin V-FITC/PI法流式细胞仪(FCM)检测细胞凋亡,用硫代巴比妥酸法测定细胞丙二醛(MDA)含量.结果 用40μmol/LtBHQ预处理后再用100、300μ,mol/L PQ处理PC12细胞24 h细胞存活率分别较100、300 μmol/L PQ组增高,差异有统计学意义(P＜0.05,P＜0.01);用40μmol/LtBHQ预处理后再用100μmol/LPQ处理PC12细胞48 h细胞存活率较100μmol/LPQ处理组细胞存活率增高,差异有统计学意义(P＜0.01);用40μmol/LtBHQ预处理后再用100、300μmol/PQ处理PC12细胞24 h后,细胞凋亡率分别较100、300 μmol/L PQ下降,差异有统计学意义(P＜0.05,P＜0.01);用40μmol/LtBHQ预处理后再用100、300μmol/LPQ处理PC12细胞24 h后,MDA含量分别较100、300 μmol/L PQ组下降,MDA含量分别为相应对照组的0.61、0.57倍,差异有统计学意义(P＜0.05).结论 tBHQ预处理可削弱PQ致PC12细胞的细胞毒性、细胞凋亡以及氧化应激作用.

Abstract：

Objective To investigate the protective effects of the tert-butylhydroquinone (tBHQ)pretreatment on neurotoxicity and oxidative stress induced by paraquat (PQ) in PC12 cells. Methods Cytoyoxicity of PC12 cells was measured by MTT assay, following the PC12 cells treatment with different concentrations of 100, 300 μmol/L PQ for 24 h and 48 h. PC12 cells were pretreated with or without 40 μmol/L tBHQ for 4 h, PC 12 cells were exposed to PQ at the doses of 0, 100, 300 μmol/L for 24 h and 48 h, respectively.The viability of PC 12 cells was measured by MTT assay, the apoptosis rates of PC 12 cells were detected by flow cytometry (FCM) and the malondialdehyde (MDA) levels of PC 12 cells were examine by thiobarbituric acid (TBA) method. Results When the exposure doses of PQ were 100 and 300 μmol/L for 24 h, the viability of PC 12 cells pretreated with tBHQ was significantly higher than that of PC 12 cells only exposed to PQ (P＜0.05 or P＜0.01). When the exposure dose of PQ was 100μmol/L for 48 h, the viability of PC12 cells pretreated with tBHQ was significantly higher than that of PC12 cells only exposed to PQ (P＜0.01). When the exposure doses of PQ were 100 and 300 μmol/L for 24 h, the apoptosis rates and MDA levels of PC12 cells pretreated with tBHQ were significantly lower than those of PC12 cells only exposed to PQ (P＜0.05 or P＜0.01). Conclusions tBHQ preteatment can reduce the cytotoxicity, apoptosis and oxidative stress induced by PQ in PC12 cells.     

叔丁基对苯二酚对亚砷酸钠致Chang liver细胞毒性的影响

目的 探讨叔丁基对苯二酚(tert-butylhydroquinone,tBHQ)对亚砷酸钠(NaAsO_2)致Chang liver细胞毒性的影响.方法 Chang liver细胞培养48 h后分别以20、40、60和80 μmo/L的NaAsO_2染毒24和48 h,作为NaAsO_2单独作用组.以5和20μmol/L的tBHQ预处理Chang liver细胞24h,以40和60μmol/L的NaAsO_2染毒24和48h,作为tBHQ预处理组;对照组处理同NaAsO_2:单独作用组.每个浓度设3个复孔.用Alamar Blue法检测细胞活力.结果 NaAsO_2单独作用24和48 h组Alamar Blue还原率显著下降,与对照组比较,差异有统计学意义(P<0.05);且NaAsO_2单独作用48 h组的Alamar Blue还原率均显著低于24 h组,差异有统计学意义(P＜0.01).5 μmol/L的tBHQ预处理组与对应的60 μmol/L NaAsO_2单独作用24h组相比,显著提高了Alamar Blue还原率(P＜0.01);20 μmol/L的tBHQ预处理组的Alamar Blue还原率均显著高于相对应NaAsO_2单独作用24 h组(P＜0.05).5 μmol/L的tBHQ预处理组的Alaraar Blue还原率均显著高于相对应NaAsO_2单独作用48h组(P＜0.01);20μmol/L的tBHQ预处理组与对应的40μmol/L NaAsO_2单独作用48h组相比,显著提高了Alamar Blue还原率(P<0.01).结论 tBHQ能够降低NaAsO_2致Chang liver细胞的毒性,增强细胞对NaAsO_2毒性的抵抗能力.

Abstract：

Objective To study the antagonism of text-butylhydroquinone (tBHQ)to NaAsO_2 induced cytotoxicity in Chang liver cells in vitro.Methods Chang liver cells were exposed to NaAsO_2(0,20,40,60 and 80μmol/L)for 24 and 48 hours,or Chang liver cells were treated with tBHQ(5 and 20 μmol/L),then exposed to NaAsO_2(40 and 60 μmol/L)for 24 and 48 hours.The conditions of control group was the same as NaAsO_2 group.Alamar Blue was used to evaluate the viabifity of cells.Results Chang liver cells were exposed to NaAsO_2 for 24 and 48 hours,Alamar Blue reduction rates decreased significantly and Alamar Blue reduction rates of 48 hours group were lower than 24 hours group (P<0.01).Alamar Blue reduction rate of 5 μmol/L tBHQ pretreatment group was higher than 60 μmol/L NaAsO_2 group for 24 h(P<0.01);Alamar Blue reduction rate of 20μmol/L tBHQ pretreatment group were higher than respective NaAsO_2 group for 24 h(P＜0.05).Alamar Blue reduction rate of 5 μmol/L tBHQ pretreatment group were higher than respective NaAsO_2 group for 48 h(P＜0.01);Alamar Blue reduction rate of 20 μmol/L tBHQ pretreatment group was higher than 40 μmol/L NaAsO_2 group for 48 h(P<0.01).Conclusion tBHQ can decrease the cytotoxieity induced by NaAsO_2 in Chang liver cells,increase the resistance to the cytotoxieity induced by NaAsO_2 in Chang liver cells.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.