中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2015年
6期
486-492
,共7页
黄婉媚%王进%黄坚毅%庞霖霖
黃婉媚%王進%黃堅毅%龐霖霖
황완미%왕진%황견의%방림림
锰中毒%5-羟色胺%脑源性神经营养因子%抑郁
錳中毒%5-羥色胺%腦源性神經營養因子%抑鬱
맹중독%5-간색알%뇌원성신경영양인자%억욱
Manganese poisoning%5-Hydroxytryptamine%Brain-derived neurotrophic factor%Depression
目的 观察慢性锰中毒大鼠脑5-羟色胺、脑源性营养因子(BDNF)的表达变化.方法 将60只雄性正常SD大鼠按完全随机数字表法分为对照组(n=15)和实验组(n=45).实验组又分为低锰组(n=15)、中锰组(n=15)、高锰组(n=15)3个亚组,每天分别腹腔注射二氯化锰四水合物5、15、25 mg/kg,每周连续注射5d,持续12周.采用蔗糖水偏好实验、敞箱实验评估大鼠是否具有抑郁样症状,采用电感耦合等离子体-原子发射光谱法测定大鼠纹状体的锰含量,高效液相色谱法测定大鼠额叶皮质、海马的5-羟色胺含量,蛋白质印记法检测大鼠额叶皮质、海马BDNF的表达,并应用实时荧光定量PCR技术检测大鼠额叶皮质、海马BDNF mRNA的相对表达量.结果 蔗糖水偏好实验及敞箱实验结果显示,慢性锰中毒大鼠均出现抑郁样行为,高锰组表现更加明显,差异有统计学意义.与对照组5-羟色胺含量(组织湿重)相比[额叶(459.65±16.81) ng/g、海马(323.92±17.41)ng/g],给锰组大鼠额叶皮质5-羟色胺含量[(423.45±17.19) ng/g、(376.89±18.87) ng/g、(280.17±25.46) ng/g]及海马中的5-羟色胺含量[(265.71±17.89) ng/g、(214.35±23.63) ng/g、(172.67±18.24) ng/g]减少,差异有统计学意义(F=132.68,P＜0.05;F =69.66,P＜0.05).与对照组相比(额叶0.962±0.111、海马0.873±0.101),给锰组大鼠BDNF的表达随锰剂量的增加而显著下降,锰浓度越高,BDNF表达水平降低越明显,给锰组各亚组大鼠之间额叶BDNF蛋白表达(0.855±0.106、0.649±0.112、0.506±0.121)、海马BDNF的表达水平(0.731±0.092、0.626±0.104、0.544±0.113)差异有统计学意义(F=13.26、P＜0.05;F=18.54,P＜0.05).与对照组相比(额叶0.000 87±0.000 07、海马0.00082±0.00009),给锰组各亚组大鼠额叶BDNF mRNA的相对表达量(0.00071±0.00006、0.000 48±0.000 03、0.00036±0.00003)及海马BDNFmRNA的表达量(0.000 57±0.000 05、0.00049±0.00004、0.00038±0.000 05)随锰剂量增加而减少,差异有统计学意义(F=18.46,P＜0.05;F=12.76,P＜0.05).结论 慢性锰中毒大鼠可出现抑郁样行为,锰蓄积可导致额叶及海马神经元内5-羟色胺、BDNF表达量下降.
目的 觀察慢性錳中毒大鼠腦5-羥色胺、腦源性營養因子(BDNF)的錶達變化.方法 將60隻雄性正常SD大鼠按完全隨機數字錶法分為對照組(n=15)和實驗組(n=45).實驗組又分為低錳組(n=15)、中錳組(n=15)、高錳組(n=15)3箇亞組,每天分彆腹腔註射二氯化錳四水閤物5、15、25 mg/kg,每週連續註射5d,持續12週.採用蔗糖水偏好實驗、敞箱實驗評估大鼠是否具有抑鬱樣癥狀,採用電感耦閤等離子體-原子髮射光譜法測定大鼠紋狀體的錳含量,高效液相色譜法測定大鼠額葉皮質、海馬的5-羥色胺含量,蛋白質印記法檢測大鼠額葉皮質、海馬BDNF的錶達,併應用實時熒光定量PCR技術檢測大鼠額葉皮質、海馬BDNF mRNA的相對錶達量.結果 蔗糖水偏好實驗及敞箱實驗結果顯示,慢性錳中毒大鼠均齣現抑鬱樣行為,高錳組錶現更加明顯,差異有統計學意義.與對照組5-羥色胺含量(組織濕重)相比[額葉(459.65±16.81) ng/g、海馬(323.92±17.41)ng/g],給錳組大鼠額葉皮質5-羥色胺含量[(423.45±17.19) ng/g、(376.89±18.87) ng/g、(280.17±25.46) ng/g]及海馬中的5-羥色胺含量[(265.71±17.89) ng/g、(214.35±23.63) ng/g、(172.67±18.24) ng/g]減少,差異有統計學意義(F=132.68,P＜0.05;F =69.66,P＜0.05).與對照組相比(額葉0.962±0.111、海馬0.873±0.101),給錳組大鼠BDNF的錶達隨錳劑量的增加而顯著下降,錳濃度越高,BDNF錶達水平降低越明顯,給錳組各亞組大鼠之間額葉BDNF蛋白錶達(0.855±0.106、0.649±0.112、0.506±0.121)、海馬BDNF的錶達水平(0.731±0.092、0.626±0.104、0.544±0.113)差異有統計學意義(F=13.26、P＜0.05;F=18.54,P＜0.05).與對照組相比(額葉0.000 87±0.000 07、海馬0.00082±0.00009),給錳組各亞組大鼠額葉BDNF mRNA的相對錶達量(0.00071±0.00006、0.000 48±0.000 03、0.00036±0.00003)及海馬BDNFmRNA的錶達量(0.000 57±0.000 05、0.00049±0.00004、0.00038±0.000 05)隨錳劑量增加而減少,差異有統計學意義(F=18.46,P＜0.05;F=12.76,P＜0.05).結論 慢性錳中毒大鼠可齣現抑鬱樣行為,錳蓄積可導緻額葉及海馬神經元內5-羥色胺、BDNF錶達量下降.
목적 관찰만성맹중독대서뇌5-간색알、뇌원성영양인자(BDNF)적표체변화.방법 장60지웅성정상SD대서안완전수궤수자표법분위대조조(n=15)화실험조(n=45).실험조우분위저맹조(n=15)、중맹조(n=15)、고맹조(n=15)3개아조,매천분별복강주사이록화맹사수합물5、15、25 mg/kg,매주련속주사5d,지속12주.채용자당수편호실험、창상실험평고대서시부구유억욱양증상,채용전감우합등리자체-원자발사광보법측정대서문상체적맹함량,고효액상색보법측정대서액협피질、해마적5-간색알함량,단백질인기법검측대서액협피질、해마BDNF적표체,병응용실시형광정량PCR기술검측대서액협피질、해마BDNF mRNA적상대표체량.결과 자당수편호실험급창상실험결과현시,만성맹중독대서균출현억욱양행위,고맹조표현경가명현,차이유통계학의의.여대조조5-간색알함량(조직습중)상비[액협(459.65±16.81) ng/g、해마(323.92±17.41)ng/g],급맹조대서액협피질5-간색알함량[(423.45±17.19) ng/g、(376.89±18.87) ng/g、(280.17±25.46) ng/g]급해마중적5-간색알함량[(265.71±17.89) ng/g、(214.35±23.63) ng/g、(172.67±18.24) ng/g]감소,차이유통계학의의(F=132.68,P＜0.05;F =69.66,P＜0.05).여대조조상비(액협0.962±0.111、해마0.873±0.101),급맹조대서BDNF적표체수맹제량적증가이현저하강,맹농도월고,BDNF표체수평강저월명현,급맹조각아조대서지간액협BDNF단백표체(0.855±0.106、0.649±0.112、0.506±0.121)、해마BDNF적표체수평(0.731±0.092、0.626±0.104、0.544±0.113)차이유통계학의의(F=13.26、P＜0.05;F=18.54,P＜0.05).여대조조상비(액협0.000 87±0.000 07、해마0.00082±0.00009),급맹조각아조대서액협BDNF mRNA적상대표체량(0.00071±0.00006、0.000 48±0.000 03、0.00036±0.00003)급해마BDNFmRNA적표체량(0.000 57±0.000 05、0.00049±0.00004、0.00038±0.000 05)수맹제량증가이감소,차이유통계학의의(F=18.46,P＜0.05;F=12.76,P＜0.05).결론 만성맹중독대서가출현억욱양행위,맹축적가도치액협급해마신경원내5-간색알、BDNF표체량하강.
Objective To observe the expression changes of 5-hydroxytryptamine (5-HT),brainderived neurotrophic factor (BDNF) in rats with chronic manganism.Methods Sixty healthy male Sprague-Dawley rats were randomly divided into control group (n =15) and experimental group (n =45).The experimental group was divided into three subgroups:low-dose group (n =15),middle-dose group (n =15),high-dose group (n =15).The rats in control group were given intraperitoneal injection of normal saline while the rats in low-dose group,middle-dose group,high-dose group were given intraperitoneal injection of 5 mg/kg,15 mg/kg and 25 mg/kg manganese chloride tetrahydrate,respectively for 5 days oncea week and lasted for 12 weeks.The depressive behavior changes of rats were observed by sucrose preference test and open field test.The concentrations of manganese in the striatum of rats were detected by inductively coupled plasma-atomic emission spectrometry.The expression of 5-HT in frontal cortex,hippocampus of rats was determined by high performance liquid chromatography.The expression of BDNF in frontal cortex,hippocampus of rats was examined by Western blotting.The expression of BDNF mRNA was detected using real-time fluorescence quantitative polymerase chain reaction.Results The chronic manganese poisoning rats presented depression-like behavior based on the sucrose preference test and open field test,which was more distinct in high-dose rats.As compared with the control group (frontal cortex (459.65 ± 16.81) ng/g,hippocampus (323.92 ± 17.41) ng/g;tissue wet weight),the expressions of 5-HT were significantly decreased in frontal cortex ((423.45 ± 17.19) ng/g,(376.89 ± 18.87) ng/g,(280.17 ± 25.46) ng/g),hippocampus ((265.71 ± 17.89) ng/g,(214.35 ±23.63) ng/g,(172.67 ± 18.24) ng/g) of the experimental group (F =132.68,69.66,both P ＜ 0.05).As compared with the control group (frontal cortex 0.962 ±0.111,hippocampus 0.873 ± 0.101;the expressions of BDNF were significantly decreased in frontal cortex (0.855 ± 0.106,0.649 ± 0.112,0.506 ± 0.121) and hippocampus (0.731 ± 0.092,0.626 ±0.104,0.544 ± 0.113) with the increasing concentration of MnCl2 which showed dose dependence (F =13.26,18.54,both P ＜ 0.05).As compared with the control group (frontal cortex 0.000 87 ± 0.000 07,hippocampus 0.000 82 ± 0.000 09),the expressions of BDNF mRNA were decreased significantly in frontal cortex (0.000 71 ± 0.000 06,0.000 48 ± 0.000 03,0.000 36 ± 0.000 03) and hippocampus (0.000 57 ± 0.000 05,0.000 49 ± 0.000 04,0.000 38 ± 0.000 05) in the treated group with the increasing concentration of manganese (F =18.46,12.76,both P ＜ 0.05).Conclusion Rats with chronic manganese poisoning could present depression-like behavior and the expression of 5-HT and BDNF is decreased in the frontal cortex and hippocampus with the increased accumulation of manganese.
