APV对大鼠海马脑片铅及谷氨酸损伤保护作用  

Protection of APV on lesions of hippocampal brain slice induced by lead and glutamate exposure in rats

作　　者：周彬[1] 文涛[2] 时胜武[1] 侯学东[1] 

机构地区：[1]沈阳医学院附属中心医院耳鼻咽喉科,辽宁沈阳110024 [2]辽宁省疾病预防控制中心

出　　处：《中国公共卫生》2016年第8期1058-1061,共4页Chinese Journal of Public Health

基　　金：辽宁省博士科研启动基金(20071063)

摘　　要：目的探讨D-2-氨基-5-膦酸基戊酸(APV)对海马脑片铅(Pb2+)及谷氨酸(Glu)损伤作用及其与脑海马细胞内钙离子浓度变化的关系。方法采用低浓度胰蛋白酶消化法分离大鼠海马细胞,实验设对照组,铅暴露、谷氨酸暴露组、APV干预组,以钙荧光探针Fura-2/AM测定海马细胞[Ca2+]i浓度;用2,3,5-氯化三苯四氮唑(TTC)染色法观察脑片损伤程度。结果 30μmol/L铅、100μmol/L谷氨酸暴露5 min后大鼠海马[Ca2+]i浓度[分别为(234.9±11.38)和(311.9±14.57)nmol/L]明显高于对照组[(109.6±5.37)nmol/L],差异有统计学意义(P<0.05);100μmol/L APV组脑海马细胞内钙离子浓度升高抑制率分别为93.4%和98.2%;30μmol/L铅、100μmol/L谷氨酸暴露30 min后大鼠海马脑片损伤率分别为(40.78±3.21)%和(44.44±3.35)%;与铅暴露组、谷氨酸暴露组比较,APV干预组海马脑片损伤率[分别为(4.05±0.67)%、(16.08±1.23)%]明显下降,差异有统计学意义(P<0.05)。结论 APV对离体海马脑片Pb2+及谷氨酸损伤具有保护作用,其机制可能与APV抑制Pb2+及谷氨酸诱发海马细胞[Ca2+]i升高、减轻钙超载的细胞毒性作用有关。Objective To explore the effect of D-（-）-2-amino-5-phosphonopentanoic acid（ APV） on lesions of hippocampal brain slice induced by lead（ Pb2 ＋） and glutamate（ Glu） and its relationship to changes of intracellular free calcium ion（ Ca2 ＋） concentration in dissociated rat hippocampal cells. Methods Lowconcentration trypsin digestion was used to dissociate hippocampal cells,and the cells were divided into control group,lead exposure group,Glu exposuregroup,and APV intervention group. The changes of intracellular free Ca2 ＋concentration was measured with Fura-2 double w avelength fluoremetry and the damage of hippocampal brain slice w as measured by means of 2,3,5-triphenyltetrazolium chloride（ TTC） dyeing. Results Five minutes after the exposure,the intracellular free Ca2 ＋concentration in dissociated rat hippocampal cells in 30 μmol / L lead and 100 μmol / L glutamate exposure group（ 234. 9 ± 11. 38 nmol / L and 311. 9± 14. 57 nmol / L） w ere significantly higher than that of the control group（ 109. 6 ± 5. 37 nmol / L）（ both P〈0. 05）. The inhibition rate of 100 μmol / L APV on the increase of intracellular free Ca2 ＋concentration in dissociated rat hippocampal cells induced by 30 μmol / L Pb2 ＋or 100 μmol / L Glu w ere 93. 4% and 98. 2%,respectively. Thirty minutes after the exposure of 30 μmol / L Pb2 ＋or 100 μmol / L Glu,the damage ratio of hippocampal slices w ere 40. 78 ± 3. 21% and 44. 44± 3. 35%. The pretreatment of 100 μmol / L APV decreased the damage ratio of hippocampal slices induced by 30 μmol /L Pb2 ＋or 100 μmol / L Glu by 4. 05 ± 0. 67% and 16. 08 ± 1. 23% compared to those of the mice w ithout the pretreatment,w ith statistically significant differences among the groups（ all P〈0. 05）. Conclusion APV has a protective effect on hippocampal slices damage induced by lead or Glu and the effect is related to the inhibition of APV on the increase of intracellular free Ca2 ＋concentration in rat hippocampal cells and the attenuation

关 键 词：D-2-氨基-5-膦酸基戊酸(APV) 细胞内游离钙[Ca2+]i 铅 谷氨酸 海马脑片 大鼠 

分 类 号：R992[医药卫生—毒理学]