中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2015年
4期
250-254
,共5页
庞霖霖%王进%黄婉媚%郭松超
龐霖霖%王進%黃婉媚%郭鬆超
방림림%왕진%황완미%곽송초
锰中毒%帕金森综合征%二价金属离子转运体%铁转运蛋白
錳中毒%帕金森綜閤徵%二價金屬離子轉運體%鐵轉運蛋白
맹중독%파금삼종합정%이개금속리자전운체%철전운단백
Manganese poisoning%Parkinsonism%Divalent metal transporter 1%Ferroportin 1
目的 研究锰中毒性帕金森综合征大鼠脑黑质二价金属离子转运体(DMT1)、铁转运蛋白(FP1)的表达变化.方法 将80只SD大鼠随机分成4组,对照组腹腔注射生理盐水,低、中、高剂量组分别腹腔注射5、15、20 mg/kg的MnCl2溶液,连续16周.第16周进行3项行为学测试.电感耦合等离子原子发射光谱法(ICP-AES)测试中脑黑质锰含量,免疫组织化学染色法观察黑质酪氨酸羟化酶(TH)阳性表达量,判定锰中毒性帕金森综合征大鼠模型是否成功;采用免疫组化及荧光定量PCR的方法测定大鼠脑黑质中DMT1、FP1的表达量.结果 该实验方法成功制备锰中毒性帕金森综合征大鼠模型.5、15、20 mg/kg染锰组大鼠中脑黑质锰含量分别为(1.72±0.33)、(2.92±0.77)、(5.65± 1.60) μg/g,均高于对照组[(0.56±0.20) μ/g],差异有统计学意义(P＜0.01).5、15、20 mg/kg染锰组中脑黑质TH阳性细胞平均吸光度值(0.054±0.008、0.016±0.004、0.003±0.001),均低于对照组(0.109±0.019),差异有统计学意义(P＜0.01).15、20 mg/kg染锰组中脑黑质DMT1阳性细胞平均吸光度值(0.062±0.004、0.116±0.064)均高于对照组(0.015±0.007),差异有统计学意义(P＜0.01).5、15、20 mg/kg染锰组中脑黑质FP1阳性细胞平均吸光度值(0.092±0.011、0.048±0.008、0.008±0.002)均低于对照组(0.306±0.081),差异有统计学意义(P＜0.01) 15、20 mg/kg染锰组大鼠黑质DMTl mRNA表达量(0.052±0.0126、0.124±0.0299)均高于对照组(0.001±0.000 4),差异有统计学意义(P＜0.05);5、15、20 mg/kg染锰组中脑黑质FP1 mRNA表达量(0.059±0.007 6,0.033±0.009 4,0.002±0.000 7)均低于对照组(0.162±0.046 3),差异有统计学意义(P＜0.05).结论 锰中毒性帕金森综合征大鼠模型中,DMT1表达增加及FP1表达降低,可能参与脑黑质锰积聚及多巴胺能神经元缺失的发生发展过程.
目的 研究錳中毒性帕金森綜閤徵大鼠腦黑質二價金屬離子轉運體(DMT1)、鐵轉運蛋白(FP1)的錶達變化.方法 將80隻SD大鼠隨機分成4組,對照組腹腔註射生理鹽水,低、中、高劑量組分彆腹腔註射5、15、20 mg/kg的MnCl2溶液,連續16週.第16週進行3項行為學測試.電感耦閤等離子原子髮射光譜法(ICP-AES)測試中腦黑質錳含量,免疫組織化學染色法觀察黑質酪氨痠羥化酶(TH)暘性錶達量,判定錳中毒性帕金森綜閤徵大鼠模型是否成功;採用免疫組化及熒光定量PCR的方法測定大鼠腦黑質中DMT1、FP1的錶達量.結果 該實驗方法成功製備錳中毒性帕金森綜閤徵大鼠模型.5、15、20 mg/kg染錳組大鼠中腦黑質錳含量分彆為(1.72±0.33)、(2.92±0.77)、(5.65± 1.60) μg/g,均高于對照組[(0.56±0.20) μ/g],差異有統計學意義(P＜0.01).5、15、20 mg/kg染錳組中腦黑質TH暘性細胞平均吸光度值(0.054±0.008、0.016±0.004、0.003±0.001),均低于對照組(0.109±0.019),差異有統計學意義(P＜0.01).15、20 mg/kg染錳組中腦黑質DMT1暘性細胞平均吸光度值(0.062±0.004、0.116±0.064)均高于對照組(0.015±0.007),差異有統計學意義(P＜0.01).5、15、20 mg/kg染錳組中腦黑質FP1暘性細胞平均吸光度值(0.092±0.011、0.048±0.008、0.008±0.002)均低于對照組(0.306±0.081),差異有統計學意義(P＜0.01) 15、20 mg/kg染錳組大鼠黑質DMTl mRNA錶達量(0.052±0.0126、0.124±0.0299)均高于對照組(0.001±0.000 4),差異有統計學意義(P＜0.05);5、15、20 mg/kg染錳組中腦黑質FP1 mRNA錶達量(0.059±0.007 6,0.033±0.009 4,0.002±0.000 7)均低于對照組(0.162±0.046 3),差異有統計學意義(P＜0.05).結論 錳中毒性帕金森綜閤徵大鼠模型中,DMT1錶達增加及FP1錶達降低,可能參與腦黑質錳積聚及多巴胺能神經元缺失的髮生髮展過程.
목적 연구맹중독성파금삼종합정대서뇌흑질이개금속리자전운체(DMT1)、철전운단백(FP1)적표체변화.방법 장80지SD대서수궤분성4조,대조조복강주사생리염수,저、중、고제량조분별복강주사5、15、20 mg/kg적MnCl2용액,련속16주.제16주진행3항행위학측시.전감우합등리자원자발사광보법(ICP-AES)측시중뇌흑질맹함량,면역조직화학염색법관찰흑질락안산간화매(TH)양성표체량,판정맹중독성파금삼종합정대서모형시부성공;채용면역조화급형광정량PCR적방법측정대서뇌흑질중DMT1、FP1적표체량.결과 해실험방법성공제비맹중독성파금삼종합정대서모형.5、15、20 mg/kg염맹조대서중뇌흑질맹함량분별위(1.72±0.33)、(2.92±0.77)、(5.65± 1.60) μg/g,균고우대조조[(0.56±0.20) μ/g],차이유통계학의의(P＜0.01).5、15、20 mg/kg염맹조중뇌흑질TH양성세포평균흡광도치(0.054±0.008、0.016±0.004、0.003±0.001),균저우대조조(0.109±0.019),차이유통계학의의(P＜0.01).15、20 mg/kg염맹조중뇌흑질DMT1양성세포평균흡광도치(0.062±0.004、0.116±0.064)균고우대조조(0.015±0.007),차이유통계학의의(P＜0.01).5、15、20 mg/kg염맹조중뇌흑질FP1양성세포평균흡광도치(0.092±0.011、0.048±0.008、0.008±0.002)균저우대조조(0.306±0.081),차이유통계학의의(P＜0.01) 15、20 mg/kg염맹조대서흑질DMTl mRNA표체량(0.052±0.0126、0.124±0.0299)균고우대조조(0.001±0.000 4),차이유통계학의의(P＜0.05);5、15、20 mg/kg염맹조중뇌흑질FP1 mRNA표체량(0.059±0.007 6,0.033±0.009 4,0.002±0.000 7)균저우대조조(0.162±0.046 3),차이유통계학의의(P＜0.05).결론 맹중독성파금삼종합정대서모형중,DMT1표체증가급FP1표체강저,가능삼여뇌흑질맹적취급다파알능신경원결실적발생발전과정.
Objective To study the changes in the expression of divalent metal transporter 1 (DMT1) and ferroportin 1 (FP1) in the substantia nigra (SN) of rats with manganese-induced parkinsonism.Methods Eighty Sprague-Dawley rats were randomly divided into four groups.Rats in the control group were injected intraperitoneally with saline solution.Rats in the low-dose,medium-dose,and high-dose groups were injected intraperitoneally with 5,15,and 20 mg/kg MnCl2 solution,respectively,for 16 weeks.Three behavioral tests were performed at the 16th week.The concentration of Mn2+ in the SN was determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES),and the positive expression of tyrosine hydroxylase (TH) was measured by immunohistochemical staining to determine whether rats with manganese-induced parkinsonism were successfully produced.The expression of DMT1 and FP1 in SN was measured by immunohistochemical staining and fluorescent quantitative polymerase chain reaction.Results Rats with manganese-induced parkinsonism were successfully produced using the above method.Compared with that in the control group,the concentrations of Mn-2+ in the SN of rats exposed to 5,15,and 20 mg/kg Mn2+ were significantly higher (1.72±0.33 vs 0.56±0.20 μg/g,P＜0.01;2.92±0.77 vs 0.56±0.20 μg/g,P＜0.01;5.65±1.60 vs 0.56±0.20 μg/g,P＜0.01).The mean ODs of TH-positive cells in the SN of rats exposed to 5,15,and 20 mg/kg Mn-2+ were significantly lower than that in the control group (0.054±0.008 vs 0.109±0.019,P＜0.01;0.016±0.004 vs 0.109±0.019,P＜0.01;0.003±0.001 vs 0.109±0.019,P＜0.01).Compared with that in the control group,the mean optical densities (ODs) of DMT1-positive cells in the SN of rats exposed to 15.and 20 mg/kg Mn2+ were significantly higher (0.062±0.004 vs 0.015±0.007,P＜0.01:0.116±0.064 vs 0.015±0.007,P＜0.01).The mean ODs of FPl-positive cells in the SN of rats exposed to 5,15,and 20 mg/kg Mn2+ were significantly lower than that in the control group (0.092±0.011 vs 0.306±0.081,P＜0.01;0.048±0.008 vs 0.306±0.081,P＜0.01:0.008±0.002 vs 0.306 ±0.081,P＜ 0.01).Rats exposed to 15 and 20 mg/kg Mn2+ had significantly higher expression of DMT1 mRNA in the SN than those in the control group (0.052±0.0126 vs 0.001±0.0004,P＜0.05;0.124±0.0299 vs 0.001±0.0004,/＜0.05).However,rats exposed to 5,15,and 20 mg/kg Mn2+ had significantly lower expression of FP1 mRNA in the SN than those in the control group (0.059±0.0076 vs 0.162±0.0463,P＜0.05;0.033±0.0094 vs 0.162±0.0463.P＜ 0.05;0.002±0.0007 vs 0.162±0.0463,P＜0.05).Conclusion The increased expression of DMT1 and reduced expression of FP1 may be involved in the processes of Mn2+ accunulation in the SN and dopaminergic nuron loss in rats with manganese-induced parkinsonism.
