中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2011年
5期
489-492
,共4页
李冰%李昕%朱博%张新玉%邢晓越%刘丹%王欣%孙贵范
李冰%李昕%硃博%張新玉%邢曉越%劉丹%王訢%孫貴範
리빙%리흔%주박%장신옥%형효월%류단%왕흔%손귀범
亚砷酸盐类%叔丁基对苯二酚%活性氧
亞砷痠鹽類%叔丁基對苯二酚%活性氧
아신산염류%숙정기대분이분%활성양
Arsenites%Tert-butylhydroquinone%Reactive oxygen species
目的 探讨叔丁基对苯二酚(tBHQ)对亚砷酸钠(NaAsO2)致细胞毒性和氧化损伤的拮抗作用。方法 Chang肝细胞用tBHQ[0(对照)、5、25μmol/L]预处理24h,再用5 μmol/L tBHQ和NaAsO2[0(对照)、30、40、50、60 μmol/L]共同作用24h,采用刃天青钠(Alamar blue)还原法检测细胞活力，结果用实验组Alamar blue还原率与对照组Alamar blue还原率的相对比值表示；Chang肝细胞用tBHQ[0(对照)、5、25 μmol/L]预处理24h,再用5μmol/L tBHQ和NaAso2[0(对照)、40、50 μmol/L]共同作用24h，采用荧光探针2′，7′-二乙酰二氯荧光素(DCFH-DA)检测细胞内活性氧(ROS)的生成，结果用实验组平均荧光强度与对照组平均荧光强度的相对比值表示。结果 30、40、50、60 μmol/L的NaAsO2暴露能够显著降低细胞活力，而tBHQ预处理(5、25 μmol/L)则可明显恢复细胞活力,NaAsO2和tBHQ两因素的主效应及其交互作用均有统计学意义（F值分别为566.57、55.09、14.50，P均＜0.05）；5、25 μmol/L tBHQ预处理的30、40、50、60 μmol/LNaAsO2组细胞活力(0.75±0.02、0.70±0.04、0.59±0.03、0.43±0.03和0.75±0.02、0.73±0.03、0.65±0.02、0.50±0.02)较相应NaAsO2单独作用组(0.70±0.03、0.64±0.03、0.43±0.03、0.33±0.01)显著升高（P均＜0.05），25 μmol/L tBHQ 预处理的50、60μmol/L NaAsO2组细胞活力高于相应5μmol/L tBHQ预处理组（P均＜0.05）。40、50 μmol/L的NaAsO2能显著诱导Chang肝细胞内ROS的产生，而tBHQ预处理(5、25 μmol/L)则可使NaAsO2诱导产生的细胞内ROS水平显著下降，NaAsO2和tBHQ两因素的主效应及其交互作用均有统计学意义（F值分别为181.78、60.55、4.93，P均＜0.05）;5、25 μmol/L tBHQ预处理的40、50 μmol/L NaAsO2组细胞内ROS水平(1.87±0.09、1.80±0.07和1.36±0.11、1.44±0.12)较相应NaAsO2单独作用组(2.30±0.18、2.18±0.17)显著降低(P 均＜ 0.05)，25 μmol/L tBHQ预处理的40、50 μmol/L NaAsO2组细胞内ROS水平低于相应5μmol/L tBHQ预处理组（P均＜ 0.05）。结论 tBHQ对NaAsO2诱导的细胞毒性和氧化损伤具有一定的拮抗作用。
目的 探討叔丁基對苯二酚(tBHQ)對亞砷痠鈉(NaAsO2)緻細胞毒性和氧化損傷的拮抗作用。方法 Chang肝細胞用tBHQ[0(對照)、5、25μmol/L]預處理24h,再用5 μmol/L tBHQ和NaAsO2[0(對照)、30、40、50、60 μmol/L]共同作用24h,採用刃天青鈉(Alamar blue)還原法檢測細胞活力，結果用實驗組Alamar blue還原率與對照組Alamar blue還原率的相對比值錶示；Chang肝細胞用tBHQ[0(對照)、5、25 μmol/L]預處理24h,再用5μmol/L tBHQ和NaAso2[0(對照)、40、50 μmol/L]共同作用24h，採用熒光探針2′，7′-二乙酰二氯熒光素(DCFH-DA)檢測細胞內活性氧(ROS)的生成，結果用實驗組平均熒光彊度與對照組平均熒光彊度的相對比值錶示。結果 30、40、50、60 μmol/L的NaAsO2暴露能夠顯著降低細胞活力，而tBHQ預處理(5、25 μmol/L)則可明顯恢複細胞活力,NaAsO2和tBHQ兩因素的主效應及其交互作用均有統計學意義（F值分彆為566.57、55.09、14.50，P均＜0.05）；5、25 μmol/L tBHQ預處理的30、40、50、60 μmol/LNaAsO2組細胞活力(0.75±0.02、0.70±0.04、0.59±0.03、0.43±0.03和0.75±0.02、0.73±0.03、0.65±0.02、0.50±0.02)較相應NaAsO2單獨作用組(0.70±0.03、0.64±0.03、0.43±0.03、0.33±0.01)顯著升高（P均＜0.05），25 μmol/L tBHQ 預處理的50、60μmol/L NaAsO2組細胞活力高于相應5μmol/L tBHQ預處理組（P均＜0.05）。40、50 μmol/L的NaAsO2能顯著誘導Chang肝細胞內ROS的產生，而tBHQ預處理(5、25 μmol/L)則可使NaAsO2誘導產生的細胞內ROS水平顯著下降，NaAsO2和tBHQ兩因素的主效應及其交互作用均有統計學意義（F值分彆為181.78、60.55、4.93，P均＜0.05）;5、25 μmol/L tBHQ預處理的40、50 μmol/L NaAsO2組細胞內ROS水平(1.87±0.09、1.80±0.07和1.36±0.11、1.44±0.12)較相應NaAsO2單獨作用組(2.30±0.18、2.18±0.17)顯著降低(P 均＜ 0.05)，25 μmol/L tBHQ預處理的40、50 μmol/L NaAsO2組細胞內ROS水平低于相應5μmol/L tBHQ預處理組（P均＜ 0.05）。結論 tBHQ對NaAsO2誘導的細胞毒性和氧化損傷具有一定的拮抗作用。
목적 탐토숙정기대분이분(tBHQ)대아신산납(NaAsO2)치세포독성화양화손상적길항작용。방법 Chang간세포용tBHQ[0(대조)、5、25μmol/L]예처리24h,재용5 μmol/L tBHQ화NaAsO2[0(대조)、30、40、50、60 μmol/L]공동작용24h,채용인천청납(Alamar blue)환원법검측세포활력，결과용실험조Alamar blue환원솔여대조조Alamar blue환원솔적상대비치표시；Chang간세포용tBHQ[0(대조)、5、25 μmol/L]예처리24h,재용5μmol/L tBHQ화NaAso2[0(대조)、40、50 μmol/L]공동작용24h，채용형광탐침2′，7′-이을선이록형광소(DCFH-DA)검측세포내활성양(ROS)적생성，결과용실험조평균형광강도여대조조평균형광강도적상대비치표시。결과 30、40、50、60 μmol/L적NaAsO2폭로능구현저강저세포활력，이tBHQ예처리(5、25 μmol/L)칙가명현회복세포활력,NaAsO2화tBHQ량인소적주효응급기교호작용균유통계학의의（F치분별위566.57、55.09、14.50，P균＜0.05）；5、25 μmol/L tBHQ예처리적30、40、50、60 μmol/LNaAsO2조세포활력(0.75±0.02、0.70±0.04、0.59±0.03、0.43±0.03화0.75±0.02、0.73±0.03、0.65±0.02、0.50±0.02)교상응NaAsO2단독작용조(0.70±0.03、0.64±0.03、0.43±0.03、0.33±0.01)현저승고（P균＜0.05），25 μmol/L tBHQ 예처리적50、60μmol/L NaAsO2조세포활력고우상응5μmol/L tBHQ예처리조（P균＜0.05）。40、50 μmol/L적NaAsO2능현저유도Chang간세포내ROS적산생，이tBHQ예처리(5、25 μmol/L)칙가사NaAsO2유도산생적세포내ROS수평현저하강，NaAsO2화tBHQ량인소적주효응급기교호작용균유통계학의의（F치분별위181.78、60.55、4.93，P균＜0.05）;5、25 μmol/L tBHQ예처리적40、50 μmol/L NaAsO2조세포내ROS수평(1.87±0.09、1.80±0.07화1.36±0.11、1.44±0.12)교상응NaAsO2단독작용조(2.30±0.18、2.18±0.17)현저강저(P 균＜ 0.05)，25 μmol/L tBHQ예처리적40、50 μmol/L NaAsO2조세포내ROS수평저우상응5μmol/L tBHQ예처리조（P균＜ 0.05）。결론 tBHQ대NaAsO2유도적세포독성화양화손상구유일정적길항작용。
Objective To study the protective effects of tert-butylhydroquinone(tBHQ) on sodium arsenite (NaAsO2)-induced cytotoxicity and oxidative injuries. Methods Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L]for 24 h, and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control),30, 40, 50, 60 μmol/L] for another 24 h, and Alamar blue reduction rates were used to evaluate cell viability,the results were expressed as the relative ratio of Alamar blue reduction rates between the experimental group and the control group. On the other hand, Chang liver cells were pretreated with tBHQ[0(control), 5, 25 μmol/L] for24 h,and then co-treated with tBHQ(5 μmol/L) together with NaAsO2[0(control), 40, 50 μmol/L] for another 24 h,and the levels of cellular reactive oxygen species(ROS) were detected by staining cells with 2',7'-dichlorofluorescin diacetate(DCFH-DA), the results were expressed as the relative ratio of mean fluorescence intensity between the experimental group and the control group. Results Cell viability decreased dramatically by treatment with NaAsO2(30, 40, 50, 60 μmol/L), while relieved to some extent by pretreatment with 5, 25 μmol/L tBHQ, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant(F =566.57, 55.09, 14.50,all P ＜ 0.05) ; the cell viability of NaAsO2(30, 40, 50, 60 μmol/L) pretreated with tBHQ(5, 25 mol/L) were 0.75 ±0.02, 0.70 ± 0.04, 0.59 ± 0.03, 0.43 ± 0.03 and 0.75 ± 0.02, 0.73 ± 0.03, 0.65 ± 0.02, 0.50 ± 0.02, respectively,all significantly higher than corresponding NaAsO2 alone groups(0.70 ± 0.03, 0.64 ± 0.03, 0.43 ± 0.03, 0.33 ±0.01, all P ＜ 0.05), the cell viability of NaAsO2(50, 60 μmol/L) pretreated with 25 μmol/L tBHQ was higher than corresponding 5 μmol/L tBHQ pretreatment groups(all P ＜ 0.05). On the other hand, 40, 50 μmol/L of NaAsO2 significantly induced hepatocellular ROS generation, while tBHQ(5, 25 μ mol/L) pretreatment significantly decreased NaAsO2-induced intracellular ROS levels, the main effects of NaAsO2 and tBHQ, as well as their interaction were all statistically significant (F =181.78, 60.55, 4.93, all P ＜ 0.05) ; the ROS levels of NaAsO2(40, 50 μ mol/L) pretreated with tBHQ(5, 25 μmol/L) were 1.87 ± 0.09, 1.80 ± 0.07 and 1.36 ± 0.11, 1.44 ± 0.12,all significantly decreased than corresponding NaAsO2 alone groups(2.30 ± 0.18, 2.18 ± 0.17, all P ＜ 0.05),the ROS levels of NaAsO2(40, 50 μmol/L) pretreated with 25 μmol/L tBHQ decreased than corresponding 5 μmol/L tBHQ pretreatment groups (all P ＜ 0.05). Conclusion tBHQ has a certain antagonism on arsenic induced cytotoxicity and oxidative injuries.
