中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2015年
6期
401-405
,共5页
王祺%张爱华%李军%唐旭东%黄晓欣
王祺%張愛華%李軍%唐旭東%黃曉訢
왕기%장애화%리군%당욱동%황효흔
砷中毒%煤%肝%氧化损伤
砷中毒%煤%肝%氧化損傷
신중독%매%간%양화손상
Arsenic poisoning%Coal%Liver%Oxidative damage
目的 观察贵州省燃煤污染型砷中毒病区砷暴露者外周血核因子E2相关因子2(Nrf2)与下游抗氧化反应元件(ARE)序列的结合能力(Nrf2-ARE结合能力)以及抗氧化酶基因的表达,为砷致机体肝氧化损伤机制的研究提供依据.方法 以贵州省兴仁县燃煤型砷中毒病区交乐村和长庆村为调查点,通过健康体检,选择161例砷暴露者为砷暴露组,按照《地方性砷中毒诊断标准》(WS/T 211-2001)分为病区非病例组(21例)和病例组(140例),病例组参照《职业性中毒性肝病诊断标准》(GBZ 59-2010)进一步分为轻度肝病组(52例)、中重度肝病组(36例),其余病例为无明显肝病组(52例);同时选择相邻的非病区上坝田村45例居民作为对照组.采集观察对象外周血提取血细胞核蛋白:凝胶电泳迁移试验(EMSA)检测其外周血细胞核内Nrf2-ARE结合能力,实时荧光定量PCR检测铜锌超氧化物歧化酶(Cu/Zn-SOD)和谷胱甘肽过氧化物酶l(GSH-Px1) mRNA相对表达量.结果 Nrf2-ARE结合能力检测结果:各组人群外周血Nrf2-ARE结合能力组间比较差异有统计学意义(F=116.033,P＜0.05).其中与对照组(3.14±1.34)比较,病区无明显肝病组(5.17±2.06)、轻度肝病组(13.13±4.84)及中重度肝病组(32.35±14.76) Nrf2-ARE结合能力均升高(P均＜0.05);与病区非病例组(5.15±3.23)及无明显肝病组比较,轻度肝病组及中重度肝病组Nrf2-ARE结合能力升高(P均＜0.05);与轻度肝病组比较,中重度肝病组Nrf2-ARE结合能力升高(P＜0.05).Cu/Zn-SOD和GSH-Px1 mRNA表达结果:病区无明显肝病组、轻度肝病组及中重度肝病组Cu/Zn-SOD mRNA相对表达量[中位数(M):1.127 8、1.257 8、1.6320]和GSH-Px1 mRNA相对表达量(M:1.334 5、1.940 9、2.062 6)均高于对照组(M:0.961 8、0.884 3),组间比较差异均有统计学意义(x2=13.065、19.934,P均＜0.05);轻度肝病组及中重度肝病组GSH-Px1 mRNA相对表达量均高于病区非病例组(M:1.248 4),组间比较差异均有统计学意义(P均＜0.05).相关性分析:砷暴露人群外周血Nrf2-ARE结合能力与Cu/Zn-SOD和GSH-Px1 mRNA表达量呈正相关(r=0.271、0.292,P均＜0.01).结论 砷所致Nrf2-ARE结合能力升高参与了砷对Nrf2下游抗氧化酶基因表达的调控,该过程可能参与了燃煤型砷中毒肝损伤的发生发展.
目的 觀察貴州省燃煤汙染型砷中毒病區砷暴露者外週血覈因子E2相關因子2(Nrf2)與下遊抗氧化反應元件(ARE)序列的結閤能力(Nrf2-ARE結閤能力)以及抗氧化酶基因的錶達,為砷緻機體肝氧化損傷機製的研究提供依據.方法 以貴州省興仁縣燃煤型砷中毒病區交樂村和長慶村為調查點,通過健康體檢,選擇161例砷暴露者為砷暴露組,按照《地方性砷中毒診斷標準》(WS/T 211-2001)分為病區非病例組(21例)和病例組(140例),病例組參照《職業性中毒性肝病診斷標準》(GBZ 59-2010)進一步分為輕度肝病組(52例)、中重度肝病組(36例),其餘病例為無明顯肝病組(52例);同時選擇相鄰的非病區上壩田村45例居民作為對照組.採集觀察對象外週血提取血細胞覈蛋白:凝膠電泳遷移試驗(EMSA)檢測其外週血細胞覈內Nrf2-ARE結閤能力,實時熒光定量PCR檢測銅鋅超氧化物歧化酶(Cu/Zn-SOD)和穀胱甘肽過氧化物酶l(GSH-Px1) mRNA相對錶達量.結果 Nrf2-ARE結閤能力檢測結果:各組人群外週血Nrf2-ARE結閤能力組間比較差異有統計學意義(F=116.033,P＜0.05).其中與對照組(3.14±1.34)比較,病區無明顯肝病組(5.17±2.06)、輕度肝病組(13.13±4.84)及中重度肝病組(32.35±14.76) Nrf2-ARE結閤能力均升高(P均＜0.05);與病區非病例組(5.15±3.23)及無明顯肝病組比較,輕度肝病組及中重度肝病組Nrf2-ARE結閤能力升高(P均＜0.05);與輕度肝病組比較,中重度肝病組Nrf2-ARE結閤能力升高(P＜0.05).Cu/Zn-SOD和GSH-Px1 mRNA錶達結果:病區無明顯肝病組、輕度肝病組及中重度肝病組Cu/Zn-SOD mRNA相對錶達量[中位數(M):1.127 8、1.257 8、1.6320]和GSH-Px1 mRNA相對錶達量(M:1.334 5、1.940 9、2.062 6)均高于對照組(M:0.961 8、0.884 3),組間比較差異均有統計學意義(x2=13.065、19.934,P均＜0.05);輕度肝病組及中重度肝病組GSH-Px1 mRNA相對錶達量均高于病區非病例組(M:1.248 4),組間比較差異均有統計學意義(P均＜0.05).相關性分析:砷暴露人群外週血Nrf2-ARE結閤能力與Cu/Zn-SOD和GSH-Px1 mRNA錶達量呈正相關(r=0.271、0.292,P均＜0.01).結論 砷所緻Nrf2-ARE結閤能力升高參與瞭砷對Nrf2下遊抗氧化酶基因錶達的調控,該過程可能參與瞭燃煤型砷中毒肝損傷的髮生髮展.
목적 관찰귀주성연매오염형신중독병구신폭로자외주혈핵인자E2상관인자2(Nrf2)여하유항양화반응원건(ARE)서렬적결합능력(Nrf2-ARE결합능력)이급항양화매기인적표체,위신치궤체간양화손상궤제적연구제공의거.방법 이귀주성흥인현연매형신중독병구교악촌화장경촌위조사점,통과건강체검,선택161례신폭로자위신폭로조,안조《지방성신중독진단표준》(WS/T 211-2001)분위병구비병례조(21례)화병례조(140례),병례조삼조《직업성중독성간병진단표준》(GBZ 59-2010)진일보분위경도간병조(52례)、중중도간병조(36례),기여병례위무명현간병조(52례);동시선택상린적비병구상패전촌45례거민작위대조조.채집관찰대상외주혈제취혈세포핵단백:응효전영천이시험(EMSA)검측기외주혈세포핵내Nrf2-ARE결합능력,실시형광정량PCR검측동자초양화물기화매(Cu/Zn-SOD)화곡광감태과양화물매l(GSH-Px1) mRNA상대표체량.결과 Nrf2-ARE결합능력검측결과:각조인군외주혈Nrf2-ARE결합능력조간비교차이유통계학의의(F=116.033,P＜0.05).기중여대조조(3.14±1.34)비교,병구무명현간병조(5.17±2.06)、경도간병조(13.13±4.84)급중중도간병조(32.35±14.76) Nrf2-ARE결합능력균승고(P균＜0.05);여병구비병례조(5.15±3.23)급무명현간병조비교,경도간병조급중중도간병조Nrf2-ARE결합능력승고(P균＜0.05);여경도간병조비교,중중도간병조Nrf2-ARE결합능력승고(P＜0.05).Cu/Zn-SOD화GSH-Px1 mRNA표체결과:병구무명현간병조、경도간병조급중중도간병조Cu/Zn-SOD mRNA상대표체량[중위수(M):1.127 8、1.257 8、1.6320]화GSH-Px1 mRNA상대표체량(M:1.334 5、1.940 9、2.062 6)균고우대조조(M:0.961 8、0.884 3),조간비교차이균유통계학의의(x2=13.065、19.934,P균＜0.05);경도간병조급중중도간병조GSH-Px1 mRNA상대표체량균고우병구비병례조(M:1.248 4),조간비교차이균유통계학의의(P균＜0.05).상관성분석:신폭로인군외주혈Nrf2-ARE결합능력여Cu/Zn-SOD화GSH-Px1 mRNA표체량정정상관(r=0.271、0.292,P균＜0.01).결론 신소치Nrf2-ARE결합능력승고삼여료신대Nrf2하유항양화매기인표체적조공,해과정가능삼여료연매형신중독간손상적발생발전.
Objective To detect the combining capacity of peripheral blood NF-E2-related factor 2 (Nrf2)of arsenic-exposed residents in the coal-contaminated arsenism area in Guizhou with the sequence of downstream antioxidant response element (ARE) as well as antioxidase gene expression,and to provide a basis for in-depth revelation of arsenic oxidative damage mechanism to human body.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey spots,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group on the basis of physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the "Diagnostic Criteria of Endemic Arsenism" (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the "Diagnostic Criteria of Occupational Chronic" (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.The hemocyte nucleoprotein was extracted from peripheral blood in the sampling subjects.The combining capacity of peripheral blood Nrf2-ARE was tested by electrophoretic mobility shift assay (EMSA),and the relative expression quantity of Cu/Zn superoxide dismutase (Cu/Zn-SOD) and glutathione peroxidase 1 (GSH-Pxl) mRNA was tested with real-time fluorescence quantification PCR (qPCR).Results The testing results of Nrf2-ARE combining capacity showed that the difference of Nrf2-ARE combining capacity between groups was statistically significant (F =116.033,P ＜ 0.05).Compared with the control group (3.14 ± 1.34),the Nrf2-ARE combining capacity was higher in the non-apparent hepatosis group (5.17 ± 2.06),mild hepatosis group (13.13 ± 4.84) and moderately severe hepatosis group (32.35 ± 14.76,all P ＜ 0.05);compared with the non-patient group (5.15 ± 3.23) and non-apparent hepatosis group,the Nrf2-ARE combining capacity of mild hepatosis group and moderately severe hepatosis group was higher (all P ＜ 0.05);compared with mild hepatosis group,the Nrf2-ARE combining capacity of moderately severe hepatosis group was higher (P ＜ 0.05).The results of Cu/Zn-SOD and GSH-Pxl mRNA expression showed the relative expression quantities of Cu/Zn-SOD [Median (M):1.127 8,1.257 8,1.632 0] and GSH-Pxl (M:1.334 5,1.940 9,2.062 6) mRNA of non-apparent hepatosis,mild hepatosis,moderately severe hepatosis groups were higher than those of the control groups (M:0.961 8,0.884 3),respectively,and their differences were statistically significant (x2 =13.065,19.934,all P ＜ 0.05).The relative expression quantities of GSH-Pxl mRNA of mild hepatosis group and moderately severe hepatosis group were higher than that of the non-patient group (M:1.248 4),and their differences were statistically significant (all P ＜ 0.05).The correlation analysis indicated the Nrf2-ARE combining capacity was positively related to the Cu/Zn-SOD and GSH-Px1 mRNA expression (r =0.271,0.292,all P ＜ 0.01).Conclusion The increase of Nrtf2-ARE combining capacity by arsenic participates is involved in the regulation of Nrf2 downstream antioxidase gene expression,and this process possibly participates in occurrence and development of coal-burning-borne arsenism and hepatic injury.
