氟、砷及其联合染毒对大鼠海马和大脑皮质组织Bcl-2和Bax蛋白表达的影响
불、신급기연합염독대대서해마화대뇌피질조직Bcl-2화Bax단백표체적영향
Effects of fluoride, arsenic and co-exposure on expression of Bcl-2 and Bax in hippocampus and cerebral cortex of rats
  • 万方数据
    中华地方病学杂志 중화지방병학잡지
    Chinese Journal of Endemiology
    2013年 4期 365-369 ,共5页

    蒋守芳%席淑华%姚三巧%佟俊旺%张艳淑%王茜%苏菁%李明艳

    장수방%석숙화%요삼교%동준왕%장염숙%왕천%소정%리명염

    氟%砷%细胞凋亡%Bcl-2相关X蛋白质 氟%砷%細胞凋亡%Bcl-2相關X蛋白質
    불%신%세포조망%Bcl-2상관X단백질
    Fluorine%Arsenic%Apoptosis%Bcl-2-associated X protein
    目的 探讨氟、砷及其联合染毒对大鼠海马和大脑皮质组织Bcl-2和Bax凋亡蛋白表达的影响.方法 将初断乳SPF级雄性SD大鼠按体质量随机分为对照组、氟处理组、砷处理组和氟砷联合组,每组10只.氟处理组大鼠饮用120 mg/L氟化钠(NaF)水溶液,砷处理组大鼠饮用70 mg/L亚砷酸钠(NaAsO2)水溶液,氟砷联合组大鼠饮用含120mg/L NaF和70 mg/L NaAsO2的水溶液,对照组大鼠饮用蒸馏水.3个月后采用免疫组织化学和Western blot法检测大鼠海马和大脑皮质组织Bcl-2和Bax凋亡蛋白的表达.结果 ①免疫组织化学法检测结果:大鼠海马和大脑皮质组织细胞质中Bcl-2和Bax凋亡蛋白呈现为棕黄色颗粒物;与对照组和氟、砷处理组比较,氟砷联合组大鼠海马和大脑皮质Bcl-2阳性神经元细胞数较少；与对照组比较,氟、砷处理组和氟砷联合组大鼠海马和大脑皮质Bax阳性神经元细胞数较多,尤其以氟砷联合组的阳性细胞数最多.②Western blot法检测结果:对照组、氟处理组、砷处理组、氟砷联合组大鼠海马组织Bcl-2、Bax蛋白表达分别为0.84±0.22、0.76±0.10、0.75±0.24、0.28±0.05和0.44±0.19、0.81±0.14、1.22±0.45、1.45±0.26,其中氟砷联合组Bcl-2表达明显低于其他3组(P均＜0.05),而氟、砷处理组和氟砷联合组Bax表达均明显高于对照组(P均＜ 0.05),且砷处理组和氟砷联合组Bax表达高于氟处理组；上述4组大鼠大脑皮质Bcl-2、Bax蛋白表达分别为0.51±0.18、0.50±0.12、0.49±0.19、0.33±0.19和0.39±0.18、0.79±0.30、0.79±0.35、0.80±0.18,其中氟、砷处理组和氟砷联合组大鼠Bax蛋白表达明显高于对照组(P均＜0.05).氟、砷处理组和氟砷联合组大鼠海马和大脑皮质组织Bcl-2/Bax表达比值(0.96±0.28、0.72±0.45、0.33±0.05和0.69±0.37、0.57±0.10、0.37±0.18)均明显低于对照组(1.91±1.32、1.44±0.29,P均＜0.05),尤以氟砷联合组最明显.结论 在一定染毒剂量下,氟和砷均促进大鼠海马及大脑皮质组织Bax蛋白的表达,降低海马及大脑皮质组织的Bcl-2/Bax表达比值.氟和砷对大鼠海马组织Bcl-2蛋白表达和大脑皮质组织Bcl-2/Bax蛋白表达比值的影响存在协同作用.
    목적 탐토불、신급기연합염독대대서해마화대뇌피질조직Bcl-2화Bax조망단백표체적영향.방법 장초단유SPF급웅성SD대서안체질량수궤분위대조조、불처리조、신처리조화불신연합조,매조10지.불처리조대서음용120 mg/L불화납(NaF)수용액,신처리조대서음용70 mg/L아신산납(NaAsO2)수용액,불신연합조대서음용함120mg/L NaF화70 mg/L NaAsO2적수용액,대조조대서음용증류수.3개월후채용면역조직화학화Western blot법검측대서해마화대뇌피질조직Bcl-2화Bax조망단백적표체.결과 ①면역조직화학법검측결과:대서해마화대뇌피질조직세포질중Bcl-2화Bax조망단백정현위종황색과립물;여대조조화불、신처리조비교,불신연합조대서해마화대뇌피질Bcl-2양성신경원세포수교소；여대조조비교,불、신처리조화불신연합조대서해마화대뇌피질Bax양성신경원세포수교다,우기이불신연합조적양성세포수최다.②Western blot법검측결과:대조조、불처리조、신처리조、불신연합조대서해마조직Bcl-2、Bax단백표체분별위0.84±0.22、0.76±0.10、0.75±0.24、0.28±0.05화0.44±0.19、0.81±0.14、1.22±0.45、1.45±0.26,기중불신연합조Bcl-2표체명현저우기타3조(P균＜0.05),이불、신처리조화불신연합조Bax표체균명현고우대조조(P균＜ 0.05),차신처리조화불신연합조Bax표체고우불처리조；상술4조대서대뇌피질Bcl-2、Bax단백표체분별위0.51±0.18、0.50±0.12、0.49±0.19、0.33±0.19화0.39±0.18、0.79±0.30、0.79±0.35、0.80±0.18,기중불、신처리조화불신연합조대서Bax단백표체명현고우대조조(P균＜0.05).불、신처리조화불신연합조대서해마화대뇌피질조직Bcl-2/Bax표체비치(0.96±0.28、0.72±0.45、0.33±0.05화0.69±0.37、0.57±0.10、0.37±0.18)균명현저우대조조(1.91±1.32、1.44±0.29,P균＜0.05),우이불신연합조최명현.결론 재일정염독제량하,불화신균촉진대서해마급대뇌피질조직Bax단백적표체,강저해마급대뇌피질조직적Bcl-2/Bax표체비치.불화신대대서해마조직Bcl-2단백표체화대뇌피질조직Bcl-2/Bax단백표체비치적영향존재협동작용.
    Objective The study was designed to explore the effects of fluoride,arsenic and co-exposure on protein expression of Bcl-2 and Bax in hippocampus and cerebral cortex tissues of rats.Methods According to body weight,forty male SPF SD rats at the beginning of weaning were randomly divided into four groups.Each group was composed of 10 rats.Rats were exposed to either 120 mg/L sodium fluoride(NaF) in fluoride treated group,70 mg/L sodium arsenite (NaAsO2) in arsenite treated group,or 120 mg/L NaF and 70 mg/L NaAsO2 combined solution in arsenic-fluoride treated group by drinking water for 3 months,respectively.The control rats drank distilled water for 3 months.The protein expressions of Bcl-2 and Bax in hippocampus and cerebral cortex of rats were detected with immunohistochemistry and Western blotting.Results ①The results of immunohistochemistry assay were as follows:The Bcl-2 and Bax protein in cytoplasm of hippocampus and cerebral cortex neurons appeared as brown particles.Compared with control,NaF-exposed and NaAsO2-exposed rats,the number of Bcl-2 positive neurons of hippocampus and cerebral cortex in co-exposed rats decreased.Compared with control rats,the number of Bax positive neurons of hippocampus and cerebral cortex in NaF-exposed,NaAsO2-exposed and coexposed rats increased,especially in co-exposed rats.②The results of Western blotting assay were as follows:the protein expression levels of Bcl-2 and Bax in hippocampus of control,NaF-exposed,NaAsO2-exposed and coexposed rats were 0.84 ± 0.22,0.76 ± 0.10,0.75 ± 0.24,0.28 ± 0.05 and 0.44 ± 0.19,0.81 ± 0.14,1.22 ± 0.45,1.45 ± 0.26,respectively.The level of Bcl-2 protein expression in hippocampus of co-exposed rats was lower than that of other three groups (all P ＜ 0.05).The levels of Bax protein expression in hippocampus in the three exposure groups were all higher than that of control rats (all P ＜ 0.05),and the levels of Bax protein expression in hippocampus of NaAsO2-exposed and co-exposed rats were higher than that of NaF-exposed rats.The protein expression levels of Bcl-2 and Bax in cerebral cortex in control,NaF-exposed,NaAsO2-exposed and coexposed rats were 0.51 ± 0.18,0.50 ± 0.12,0.49 ± 0.19,0.33 ± 0.19 and 0.39 ± 0.18,0.79 ± 0.30,0.79 ± 0.35,0.80 ± 0.18,respectively.Compared with control rats,the Bax protein expressions in cerebral cortex of rats exposed to NaF,NaAsO2 and their combined solution were all higher(all P ＜ 0.05).We also found that the ratios of Bcl-2 and Bax expression in hippocampus and cerebral cortex of NaF-exposed (0.96 ± 0.28,0.69 ± 0.37),NaAsO2-exposed(0.72 ± 0.45,0.57 ± 0.10) and co-exposed rats(0.33 ± 0.05,0.37± 0.18) all markedly decreased than that of control rats(1.91 ± 1.32,1.44 ± 0.29,all P ＜ 0.05),especially in co-exposed rats.Conclusions Under certain exposure doses,fluoride and arsenic have increased Bax protein expression,decreased Bcl-2/Bax expression ratio in hippocampus and cerebral cortex of rats.Fluoride and arsenic might have synergistic effect on Bcl-2 protein expression in hippocampus and Bcl-2/Bax expression ratio in cerebral cortex of rats.
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