中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2009年
7期
622-627
,共6页
刘利%金一和%王烈%于红瑶%刘薇%于棋麟%王柯%刘冰%王静
劉利%金一和%王烈%于紅瑤%劉薇%于棋麟%王柯%劉冰%王靜
류리%금일화%왕렬%우홍요%류미%우기린%왕가%류빙%왕정
烷烃类%磺酸类%大脑皮质%海马%受体,N-甲基-D-天冬氨酸
烷烴類%磺痠類%大腦皮質%海馬%受體,N-甲基-D-天鼕氨痠
완경류%광산류%대뇌피질%해마%수체,N-갑기-D-천동안산
Alkanes%Sulfonic acids%Cerebral cortex%Hippecampus%Receptors,N-methyl-D-aspartate
目的 观察出生前后全氟辛烷磺酸(PFOS)暴露对大鼠仔鼠空间学习记忆能力以及大脑皮质和海马结构中N-甲基-D-门冬氨酸受体2B(NR2B)亚单位mRNA和蛋白水平的影响,探讨PFOS所致神经发育毒性机制.方法 应用数字表法将28只受孕Wistar大鼠按3:2:2比例随机分配到对照(C)、低剂量(L)和高剂量(H)组,大鼠从妊娠第0天开始分别自由摄食含0、7.2、14.4 mg/kg PFOS的粉末状饲料进行连续染毒至仔鼠出生后30 d.采用交叉哺育的方法,建立仔鼠出生前后均不暴露(CC)、仅出生前暴露(LC和HC)、仅出生后暴露(CL和CH)、出生前后均暴露(LL和HH)于PFOS的动物模型.水迷宫实验检测仔鼠空间学习和记忆能力,半定量反转录聚合酶链反应(RT-PCR)法检测仔鼠大脑额叶皮质NR2B mRNA水平,免疫组织化学染色法观察仔鼠大脑皮质(额叶及颞叶皮质)和海马组织(CA1、CA3、CA4和DG区)中NR2B蛋白的表达情况.结果 水迷宫实验洲练第4天时CL、CH、LL和HH组仔鼠逃避潜伏期分别为(99.83±25.77)s、(111.30±17.82)s、(106.40±18.71)s、(107.70±16.85)s,显著长于CC组[(54.90±26.69)s](q值分别为4.349、4.773、6.026、5.641,P值均<0.01);训练第4天时HH组仔鼠进入盲端的错误次数为(22.30±7.56)次,显著高于Cc组[(9.80±4.64)次](q=5.173,P<0.01).出生后第l天(postnatal day 1,PND1)HC组和PND14时LC组、HC组、HH组仔鼠大脑额叶皮质中NR2B mRNA水平分别为(0.167±0.008)、(0.364±0.035)、(0.341±0.030)、(0.328±0.045),均显著低于CC组的(0.271±0.060)和(0.465±0.067),差异有统计学意义(q值分别为3.547、3.739、4.597、5.006,P值均<0.05).PNDI时LC组仔鼠海马CA1区NR2B蛋白水平为(0.091±0.005),显著低于CC组(0.123±0.009)(q=5.209,P<0.05);PND14时,PFOS的影响扩大到仔鼠大脑皮质和海马结构各区;PND28时PFOS的影响见于CA1、CA3和颞叶皮质区.结论 出生前后不同时期PFOS暴露导致大鼠仔鼠空间学习和记忆能力损伤,其机制可能为大脑皮质和海马结构各区NR2B表达水平的降低.
目的 觀察齣生前後全氟辛烷磺痠(PFOS)暴露對大鼠仔鼠空間學習記憶能力以及大腦皮質和海馬結構中N-甲基-D-門鼕氨痠受體2B(NR2B)亞單位mRNA和蛋白水平的影響,探討PFOS所緻神經髮育毒性機製.方法 應用數字錶法將28隻受孕Wistar大鼠按3:2:2比例隨機分配到對照(C)、低劑量(L)和高劑量(H)組,大鼠從妊娠第0天開始分彆自由攝食含0、7.2、14.4 mg/kg PFOS的粉末狀飼料進行連續染毒至仔鼠齣生後30 d.採用交扠哺育的方法,建立仔鼠齣生前後均不暴露(CC)、僅齣生前暴露(LC和HC)、僅齣生後暴露(CL和CH)、齣生前後均暴露(LL和HH)于PFOS的動物模型.水迷宮實驗檢測仔鼠空間學習和記憶能力,半定量反轉錄聚閤酶鏈反應(RT-PCR)法檢測仔鼠大腦額葉皮質NR2B mRNA水平,免疫組織化學染色法觀察仔鼠大腦皮質(額葉及顳葉皮質)和海馬組織(CA1、CA3、CA4和DG區)中NR2B蛋白的錶達情況.結果 水迷宮實驗洲練第4天時CL、CH、LL和HH組仔鼠逃避潛伏期分彆為(99.83±25.77)s、(111.30±17.82)s、(106.40±18.71)s、(107.70±16.85)s,顯著長于CC組[(54.90±26.69)s](q值分彆為4.349、4.773、6.026、5.641,P值均<0.01);訓練第4天時HH組仔鼠進入盲耑的錯誤次數為(22.30±7.56)次,顯著高于Cc組[(9.80±4.64)次](q=5.173,P<0.01).齣生後第l天(postnatal day 1,PND1)HC組和PND14時LC組、HC組、HH組仔鼠大腦額葉皮質中NR2B mRNA水平分彆為(0.167±0.008)、(0.364±0.035)、(0.341±0.030)、(0.328±0.045),均顯著低于CC組的(0.271±0.060)和(0.465±0.067),差異有統計學意義(q值分彆為3.547、3.739、4.597、5.006,P值均<0.05).PNDI時LC組仔鼠海馬CA1區NR2B蛋白水平為(0.091±0.005),顯著低于CC組(0.123±0.009)(q=5.209,P<0.05);PND14時,PFOS的影響擴大到仔鼠大腦皮質和海馬結構各區;PND28時PFOS的影響見于CA1、CA3和顳葉皮質區.結論 齣生前後不同時期PFOS暴露導緻大鼠仔鼠空間學習和記憶能力損傷,其機製可能為大腦皮質和海馬結構各區NR2B錶達水平的降低.
목적 관찰출생전후전불신완광산(PFOS)폭로대대서자서공간학습기억능력이급대뇌피질화해마결구중N-갑기-D-문동안산수체2B(NR2B)아단위mRNA화단백수평적영향,탐토PFOS소치신경발육독성궤제.방법 응용수자표법장28지수잉Wistar대서안3:2:2비례수궤분배도대조(C)、저제량(L)화고제량(H)조,대서종임신제0천개시분별자유섭식함0、7.2、14.4 mg/kg PFOS적분말상사료진행련속염독지자서출생후30 d.채용교차포육적방법,건립자서출생전후균불폭로(CC)、부출생전폭로(LC화HC)、부출생후폭로(CL화CH)、출생전후균폭로(LL화HH)우PFOS적동물모형.수미궁실험검측자서공간학습화기억능력,반정량반전록취합매련반응(RT-PCR)법검측자서대뇌액협피질NR2B mRNA수평,면역조직화학염색법관찰자서대뇌피질(액협급섭협피질)화해마조직(CA1、CA3、CA4화DG구)중NR2B단백적표체정황.결과 수미궁실험주련제4천시CL、CH、LL화HH조자서도피잠복기분별위(99.83±25.77)s、(111.30±17.82)s、(106.40±18.71)s、(107.70±16.85)s,현저장우CC조[(54.90±26.69)s](q치분별위4.349、4.773、6.026、5.641,P치균<0.01);훈련제4천시HH조자서진입맹단적착오차수위(22.30±7.56)차,현저고우Cc조[(9.80±4.64)차](q=5.173,P<0.01).출생후제l천(postnatal day 1,PND1)HC조화PND14시LC조、HC조、HH조자서대뇌액협피질중NR2B mRNA수평분별위(0.167±0.008)、(0.364±0.035)、(0.341±0.030)、(0.328±0.045),균현저저우CC조적(0.271±0.060)화(0.465±0.067),차이유통계학의의(q치분별위3.547、3.739、4.597、5.006,P치균<0.05).PNDI시LC조자서해마CA1구NR2B단백수평위(0.091±0.005),현저저우CC조(0.123±0.009)(q=5.209,P<0.05);PND14시,PFOS적영향확대도자서대뇌피질화해마결구각구;PND28시PFOS적영향견우CA1、CA3화섭협피질구.결론 출생전후불동시기PFOS폭로도치대서자서공간학습화기억능력손상,기궤제가능위대뇌피질화해마결구각구NR2B표체수평적강저.
Objective To study the effects of prenatal and postnatal perfluorooetane sulfonate (PFOS) exposure on spatial learning and memory, N-methyl-D-aspartate receptor 2B (NR2B) mRNA and protein level in frontal cortex and hippoeampus of rat pups and to explore the mechanism of developmental neurotoxicity induced by PFOS. Methods Twenty-eight pregnant mrs were randomly divided into three groups in proportion of 3: 2: 2,including control group (C), low dose group (L) and high dose group (H) by means of randomized number table, which respectively received 0,7.2, 14.4 mg/kg PFOS feed from pregnancy day 0 to postnatal day(PND)30 by free feedings. The animal models of prenatal and postnatal non-exposure (CC), prenatal exposure (LC and HC), postnatal exposure (CL and CH), and prenatal and postnatal exposure (LL and HH) to PFOS were established by cross-fostering method. The spatial learning and memory were measured by water maze experiment,the NR2B mRNA levels in frontal cortex of rat pups was determined with semi-quantitative RT-PCR, NR2B protein express in cerebral cortex (frontal and tempera]cortex) and hippocampus (CAI, CA3, CA4 and DG regions) of rat pups was detected by immunohistochemistry. Results The escape latency of CL, CH, LL and HH groups pups in water maze experiment were (99. 83±25.77) s, (111.30±17. 82) s, (106. 40±18. 71) s, (107.70±16. 85) s, and longer as compared with CC group[(54.90±26.69)s](q value were 4.349,4.773,6.026 and 5.641, respectively,P<0. 01). The number of errors of HH group rat pups entering dead end was (22. 30±7.56) at the training day 4,and it was significantly higher than that of CC group (9. 80±4. 64) (q=5. 173,P<0. 01). The NR2B mRNA levels of frontal cortex of pups in HC group at PND1 ,and LC group, HC group and HH group at PND14 were (0. 167±0. 008), (0. 364±0. 035), (0. 341±0. 030) and (0. 328±0. 045) respectively,which were significantly lower than CC group (0.271±0.060) and (0.465±0.067) (q values were 3. 547, 3. 739, 4. 597 and 5. 006, respectively, P<0.05) . The results of immunohistochemistry indicated that NR2B protein express of the hippocampus CA1 region of pups in LC group was (0.091±0.005),and showed significant lower than CC group which was (0. 123±0.009) at PND1 (q=5. 209 ,P<0. 05). At PND14, the effect of PFOS extended to cerebral cortex and hippocampus regions. At PND28, the effects of PFOS were showed in hippocampus CA1, CA3 and temporal cortex regions. Conclusion Prenatal and postnatal exposure to PFOS should result in the spatial learning and memory damage,and the mechanism might be possibly involved in the decrease of NR2B level in cerebra lcortex and hippocampal formation regions.
