目的 观察硫氧还蛋白还原酶(TrxR)1在燃煤污染型砷中毒病区砷暴露人群和大鼠血液和肝脏中的表达,探讨其在燃煤污染型砷中毒肝损伤中的作用.方法 ①人群研究:选择贵州省燃煤污染型砷中毒病区133例砷暴露者作为观察对象,依据临床检查和诊断结果分为病区非病人组(25例)、无明显肝病组(38例)、轻度(43例)和中重度肝病组(27例);以非砷污染村34名健康居民为对照组.采集上述观察对象的外周血,采用实时荧光定量PCR法检测TrxR1 mRNA表达,并用可见分光光度法检测TrxR酶活力.②动物实验:将30只体质量为80 ～ 100 g的Wiser大鼠,采用随机数字表法按体质量分为对照组、饮水型砷中毒组和低、中、高砷粮食污染组,共5组,每组6只.对照组常规喂养,饮水型砷中毒组饮用含10 mg/kg三氧化二砷(As2O3)的水溶液,低、中、高砷粮食污染组喂饲用病区高砷煤烘烤的玉米粉所配制的饲料(含砷量分别为25、50、100 mg/kg),均喂养3个月.采用实时荧光定量PCR法检测其外周血和肝组织中TrxR1 mRNA表达,免疫组织化学法检测肝组织TrxR1蛋白表达,可见分光光度法检测血清和肝组织中TrxR酶活力.结果 ①人群研究结果:病区非病人组、无明显肝病组、轻度和中重度肝病组外周血TrxR1 mRNA表达[中位数(四分位数)]分别为1.599 8(1.128 9～2.156 8)、1.469 3(1.146 1～1.976 3)、1.203 6(0.463 1～1.816 2)和0.912 3(0.631 8～1.535 0),轻度和中重度肝病组均低于对照组[1.649 7(1.161 1～2.380 2),P均＜0.05];血清TrxR酶活力分别为(3.12±0.76)、(2.81±0.84)、(2.52±0.73)、(2.42±0.76)kU/L,轻度和中重度肝病组均低于对照组[(3.02±0.70)kU/L,P均＜0.05].②动物实验结果:大鼠饮水型砷中毒组和低、中、高砷粮食污染组外周血TrxR1 mRNA表达分别为1.05±0.14、1.18±0.18、1.04±0.10、0.97±0.13,除低砷粮食污染组外,其余各染砷组均低于对照组(1.23±0.15,P均＜0.05);肝组织TrxR1 mRNA表达分别为0.78±0.10、0.83±0.10、0.79±0.09、0.77±0.11,除低砷粮食污染组外,其余各染砷组均低于对照组(0.94±0.12,P均＜0.05);肝组织TrxR1蛋白表达量分别为310.33±38.81、312.50±23.36、305.67±20.57、298.17±23.52,各染砷组均低于对照组(348.50±32.35,P均＜0.05);血清TrxR酶活力分别为(4.22±0.73)、(4.86±0.63)、(4.04±0.57)、(3.73±0.64)kU/L,各染砷组均低于对照组[(9.52±1.08)kU/L,P均＜0.05];肝组织TrxR酶活力分别为(14.82±1.67)、(18.76±2.76)、(14.90±2.17)、(11.55±1.74)U/mg,均低于对照组[(23.71±3.05)U/mg,P均＜0.05].结论 砷通过下调TrxR1转录表达和蛋白表达,降低其酶活力,加重燃煤污染型砷中毒肝损伤的发生发展.
目的 觀察硫氧還蛋白還原酶(TrxR)1在燃煤汙染型砷中毒病區砷暴露人群和大鼠血液和肝髒中的錶達,探討其在燃煤汙染型砷中毒肝損傷中的作用.方法 ①人群研究:選擇貴州省燃煤汙染型砷中毒病區133例砷暴露者作為觀察對象,依據臨床檢查和診斷結果分為病區非病人組(25例)、無明顯肝病組(38例)、輕度(43例)和中重度肝病組(27例);以非砷汙染村34名健康居民為對照組.採集上述觀察對象的外週血,採用實時熒光定量PCR法檢測TrxR1 mRNA錶達,併用可見分光光度法檢測TrxR酶活力.②動物實驗:將30隻體質量為80 ～ 100 g的Wiser大鼠,採用隨機數字錶法按體質量分為對照組、飲水型砷中毒組和低、中、高砷糧食汙染組,共5組,每組6隻.對照組常規餵養,飲水型砷中毒組飲用含10 mg/kg三氧化二砷(As2O3)的水溶液,低、中、高砷糧食汙染組餵飼用病區高砷煤烘烤的玉米粉所配製的飼料(含砷量分彆為25、50、100 mg/kg),均餵養3箇月.採用實時熒光定量PCR法檢測其外週血和肝組織中TrxR1 mRNA錶達,免疫組織化學法檢測肝組織TrxR1蛋白錶達,可見分光光度法檢測血清和肝組織中TrxR酶活力.結果 ①人群研究結果:病區非病人組、無明顯肝病組、輕度和中重度肝病組外週血TrxR1 mRNA錶達[中位數(四分位數)]分彆為1.599 8(1.128 9～2.156 8)、1.469 3(1.146 1～1.976 3)、1.203 6(0.463 1～1.816 2)和0.912 3(0.631 8～1.535 0),輕度和中重度肝病組均低于對照組[1.649 7(1.161 1～2.380 2),P均＜0.05];血清TrxR酶活力分彆為(3.12±0.76)、(2.81±0.84)、(2.52±0.73)、(2.42±0.76)kU/L,輕度和中重度肝病組均低于對照組[(3.02±0.70)kU/L,P均＜0.05].②動物實驗結果:大鼠飲水型砷中毒組和低、中、高砷糧食汙染組外週血TrxR1 mRNA錶達分彆為1.05±0.14、1.18±0.18、1.04±0.10、0.97±0.13,除低砷糧食汙染組外,其餘各染砷組均低于對照組(1.23±0.15,P均＜0.05);肝組織TrxR1 mRNA錶達分彆為0.78±0.10、0.83±0.10、0.79±0.09、0.77±0.11,除低砷糧食汙染組外,其餘各染砷組均低于對照組(0.94±0.12,P均＜0.05);肝組織TrxR1蛋白錶達量分彆為310.33±38.81、312.50±23.36、305.67±20.57、298.17±23.52,各染砷組均低于對照組(348.50±32.35,P均＜0.05);血清TrxR酶活力分彆為(4.22±0.73)、(4.86±0.63)、(4.04±0.57)、(3.73±0.64)kU/L,各染砷組均低于對照組[(9.52±1.08)kU/L,P均＜0.05];肝組織TrxR酶活力分彆為(14.82±1.67)、(18.76±2.76)、(14.90±2.17)、(11.55±1.74)U/mg,均低于對照組[(23.71±3.05)U/mg,P均＜0.05].結論 砷通過下調TrxR1轉錄錶達和蛋白錶達,降低其酶活力,加重燃煤汙染型砷中毒肝損傷的髮生髮展.
목적 관찰류양환단백환원매(TrxR)1재연매오염형신중독병구신폭로인군화대서혈액화간장중적표체,탐토기재연매오염형신중독간손상중적작용.방법 ①인군연구:선택귀주성연매오염형신중독병구133례신폭로자작위관찰대상,의거림상검사화진단결과분위병구비병인조(25례)、무명현간병조(38례)、경도(43례)화중중도간병조(27례);이비신오염촌34명건강거민위대조조.채집상술관찰대상적외주혈,채용실시형광정량PCR법검측TrxR1 mRNA표체,병용가견분광광도법검측TrxR매활력.②동물실험:장30지체질량위80 ～ 100 g적Wiser대서,채용수궤수자표법안체질량분위대조조、음수형신중독조화저、중、고신양식오염조,공5조,매조6지.대조조상규위양,음수형신중독조음용함10 mg/kg삼양화이신(As2O3)적수용액,저、중、고신양식오염조위사용병구고신매홍고적옥미분소배제적사료(함신량분별위25、50、100 mg/kg),균위양3개월.채용실시형광정량PCR법검측기외주혈화간조직중TrxR1 mRNA표체,면역조직화학법검측간조직TrxR1단백표체,가견분광광도법검측혈청화간조직중TrxR매활력.결과 ①인군연구결과:병구비병인조、무명현간병조、경도화중중도간병조외주혈TrxR1 mRNA표체[중위수(사분위수)]분별위1.599 8(1.128 9～2.156 8)、1.469 3(1.146 1～1.976 3)、1.203 6(0.463 1～1.816 2)화0.912 3(0.631 8～1.535 0),경도화중중도간병조균저우대조조[1.649 7(1.161 1～2.380 2),P균＜0.05];혈청TrxR매활력분별위(3.12±0.76)、(2.81±0.84)、(2.52±0.73)、(2.42±0.76)kU/L,경도화중중도간병조균저우대조조[(3.02±0.70)kU/L,P균＜0.05].②동물실험결과:대서음수형신중독조화저、중、고신양식오염조외주혈TrxR1 mRNA표체분별위1.05±0.14、1.18±0.18、1.04±0.10、0.97±0.13,제저신양식오염조외,기여각염신조균저우대조조(1.23±0.15,P균＜0.05);간조직TrxR1 mRNA표체분별위0.78±0.10、0.83±0.10、0.79±0.09、0.77±0.11,제저신양식오염조외,기여각염신조균저우대조조(0.94±0.12,P균＜0.05);간조직TrxR1단백표체량분별위310.33±38.81、312.50±23.36、305.67±20.57、298.17±23.52,각염신조균저우대조조(348.50±32.35,P균＜0.05);혈청TrxR매활력분별위(4.22±0.73)、(4.86±0.63)、(4.04±0.57)、(3.73±0.64)kU/L,각염신조균저우대조조[(9.52±1.08)kU/L,P균＜0.05];간조직TrxR매활력분별위(14.82±1.67)、(18.76±2.76)、(14.90±2.17)、(11.55±1.74)U/mg,균저우대조조[(23.71±3.05)U/mg,P균＜0.05].결론 신통과하조TrxR1전록표체화단백표체,강저기매활력,가중연매오염형신중독간손상적발생발전.
Objective To study the expression and enzyme activity of thioredoxin reductase 1 (TrxR1) in liver and peripheral blood of human and rats exposed to airborne arsenic through coal-burning as well as its role in liver injury of coal-burning-borne arsenic poisoning.Methods This study was divided into 2 parts.Part 1 was a population study:133 local residents exposed to airborne arsenic through coal-burning were selected as arsenic exposure groups including a non-patient group (25 cases),no obvious hepatopathy group (38 cases),mild (43 cases) and moderate to severe hepatopathy groups (27 cases) from areas affected by endemic arsenism in Guizhou Province.Thirty-four healthy residents from arsenic not affected areas were selected as controls.Peripheral blood samples were collected from all these people.The expression of TrxR1 mRNA was determined by real-time fluorescence quantitative PCR (qPCR),and enzyme activity of TrxR was tested by visible spectrophotometry.Part 2 was an animal experiment study:Thirty Wistar rats,weighing about 80-100 g,were divided into control group,drinking-waterborne arsenic poisoning group and coal-burning-borne arsenic poisoning group (including low,medium and high arsenic contaminated grain groups) by means of a table of random number according to body mass,6 rats in each group.The control group was fed with normal diet for 3 months; drinking-water-borne arsenic poisoning group and coal-burning-borne arsenic poisoning group were fed with 10 mg/kg As2O3 solution and different concentrations(25,50,100 mg/kg) of arsenic-containing feed,respectively,for 3 months.The expression of TrxR1 mRNA was determined by qPCR; protein expression level of TrxR1 in liver tissue was detected by immunohistochemistry,and enzyme activity of TrxR in serum and liver tissue was tested by visible spectrophotometry.Results The mRNA expressions of TrxR1 in peripheral blood were 1.599 8 (1.128 9-2.156 8),1.469 3 (1.146 1-1.976 3),1.203 6 (0.463 1-1.816 2) and 0.912 3(0.631 8-1.535 0),respectively,among non-patient group,no obvious hepatopathy group,mild and moderate to severe hepatopathy groups.Compared to the control group[1.649 7(1.161 1-2.380 2)],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The enzyme activity of TrxR in peripheral blood was (3.12 ± 0.76),(2.81 ± 0.84),(2.52 ± 0.73),(2.42 ± 0.76)U/ml,respectively,in those corresponding groups.Compared to the control group [(3.02 ± 0.70)U/ml],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The mRNA expressions of TrxR1 in peripheral blood were 1.05 ± 0.14,1.18 ± 0.18,1.04 ± 0.10 and 0.97 ± 0.13,respectively,among drinking-water-borne arsenic poisoning group,low,medium and high arsenic contaminated grain groups; all of which were lower than that in the control group (1.23 ± 0.15,all P ＜ 0.05) except that of the low arsenic contaminated grain group.The mRNA expressions of TrxR1 in liver tissue were 0.78± 0.10,0.83 ± 0.10,0.79 ± 0.09 and 0.77 ± 0.11,respectively; all of which were lower than that in the control group (0.94 ± 0.12,all P ＜ 0.05).The protein expression of TrxR1 in liver tissue was 310.33 ± 38.81,312.50 ± 23.36,305.67 ± 20.57 and 298.17 ± 23.52,respectively,among the arsenic poisoning groups; all of which were lower than that in the control group (348.50 ± 32.35,all P ＜ 0.05).The enzyme activity of TrxR in serum was (4.22 ± 0.73),(4.86 ± 0.63),(4.04 ± 0.57),(3.73 ± 0.64)U/ml,respectively; all of which were lower than that in the control group [(9.52 ± 1.08)U/ml,all P ＜ 0.05].The enzyme activity of TrxR in liver tissue was (14.82 ± 1.67),(18.76 ± 2.76),(14.90 ± 2.17),(11.55 ± 1.74) U/mg,respectively; all of which were lower than that in the control group [(23.71 ± 3.05)U/mg,all P ＜ 0.05].Conclusion Arsenic aggravates liver injury of coal-burning arsenic poisoning through down-regulating the expressions of TrxR1 mRNA and protein and reducing its enzyme activity as well.
