中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2015年
1期
22-24
,共3页
王艳%戴莉莉%赵凤红%金亚平
王豔%戴莉莉%趙鳳紅%金亞平
왕염%대리리%조봉홍%금아평
砷%细胞%钙
砷%細胞%鈣
신%세포%개
Arsenite%Cells%Calcium
目的 观察砷暴露对大鼠原代星形胶质细胞(astrocyte,AST)分泌胶质细胞源性递质的影响,探讨砷对学习记忆功能损伤的作用机制.方法 出生1～3 d的Wistar大鼠仔鼠,取双侧大脑半球,经处理获得原代培养脑AST,并通过胶质纤维酸性蛋白免疫荧光染色鉴定.AST分别在含0.0、2.5、5.0、10.0μmol/L亚砷酸钠的培养液中培养24h,荧光双波长分光光度计法检测细胞内游离钙离子浓度([Ca2+]i);高效液相色谱(HPLC)法检测细胞培养液谷氨酸、D-丝氨酸、甘氨酸和γ-氨基丁酸含量.结果 胶质纤维酸性蛋白免疫荧光染色,AST纯度＞95％.不同浓度砷暴露组间AST内[Ca2+]i比较差异有统计学意义(F=20.030,P＜0.05),其中10.0 μmol/L砷暴露组AST内[Ca2+]i[(263.27±14.80)nmol/L]明显高于0.0、2.5、5.0μmol/L砷暴露组[(204.24±27.21)、(214.49±21.85)、(232.74±23.14)nmol/L,P均＜0.05].不同浓度砷暴露组间AST分泌的D-丝氨酸、甘氨酸和γ-氨基丁酸比较差异均有统计学意义(F值分别为26.599、33.539、5.599,P均＜0.05),其中2.5、5.0、10.0 μmol/L砷暴露组AST分泌的D-丝氨酸[(21.580±1.313)、(21.936±1.539)、(23.401±1.648)μmol/L]、甘氨酸[(26.353±2.449)、(29.711±1.530)、(29.234±2.057)μmol/L]和γ-氨基丁酸[(27.277±3.421)、(30.213±2.098)、(29.364±2.588) μmol/L]均高于0.0 μmol/L砷暴露组[(16.017±1.046)、(16.763±3.007)、(22.736±4.139) μmol/L,P均＜0.05].结论 砷暴露可引起原代AST分泌胶质细胞源性递质增加,可能会损伤学习记忆功能.
目的 觀察砷暴露對大鼠原代星形膠質細胞(astrocyte,AST)分泌膠質細胞源性遞質的影響,探討砷對學習記憶功能損傷的作用機製.方法 齣生1～3 d的Wistar大鼠仔鼠,取雙側大腦半毬,經處理穫得原代培養腦AST,併通過膠質纖維痠性蛋白免疫熒光染色鑒定.AST分彆在含0.0、2.5、5.0、10.0μmol/L亞砷痠鈉的培養液中培養24h,熒光雙波長分光光度計法檢測細胞內遊離鈣離子濃度([Ca2+]i);高效液相色譜(HPLC)法檢測細胞培養液穀氨痠、D-絲氨痠、甘氨痠和γ-氨基丁痠含量.結果 膠質纖維痠性蛋白免疫熒光染色,AST純度＞95％.不同濃度砷暴露組間AST內[Ca2+]i比較差異有統計學意義(F=20.030,P＜0.05),其中10.0 μmol/L砷暴露組AST內[Ca2+]i[(263.27±14.80)nmol/L]明顯高于0.0、2.5、5.0μmol/L砷暴露組[(204.24±27.21)、(214.49±21.85)、(232.74±23.14)nmol/L,P均＜0.05].不同濃度砷暴露組間AST分泌的D-絲氨痠、甘氨痠和γ-氨基丁痠比較差異均有統計學意義(F值分彆為26.599、33.539、5.599,P均＜0.05),其中2.5、5.0、10.0 μmol/L砷暴露組AST分泌的D-絲氨痠[(21.580±1.313)、(21.936±1.539)、(23.401±1.648)μmol/L]、甘氨痠[(26.353±2.449)、(29.711±1.530)、(29.234±2.057)μmol/L]和γ-氨基丁痠[(27.277±3.421)、(30.213±2.098)、(29.364±2.588) μmol/L]均高于0.0 μmol/L砷暴露組[(16.017±1.046)、(16.763±3.007)、(22.736±4.139) μmol/L,P均＜0.05].結論 砷暴露可引起原代AST分泌膠質細胞源性遞質增加,可能會損傷學習記憶功能.
목적 관찰신폭로대대서원대성형효질세포(astrocyte,AST)분비효질세포원성체질적영향,탐토신대학습기억공능손상적작용궤제.방법 출생1～3 d적Wistar대서자서,취쌍측대뇌반구,경처리획득원대배양뇌AST,병통과효질섬유산성단백면역형광염색감정.AST분별재함0.0、2.5、5.0、10.0μmol/L아신산납적배양액중배양24h,형광쌍파장분광광도계법검측세포내유리개리자농도([Ca2+]i);고효액상색보(HPLC)법검측세포배양액곡안산、D-사안산、감안산화γ-안기정산함량.결과 효질섬유산성단백면역형광염색,AST순도＞95％.불동농도신폭로조간AST내[Ca2+]i비교차이유통계학의의(F=20.030,P＜0.05),기중10.0 μmol/L신폭로조AST내[Ca2+]i[(263.27±14.80)nmol/L]명현고우0.0、2.5、5.0μmol/L신폭로조[(204.24±27.21)、(214.49±21.85)、(232.74±23.14)nmol/L,P균＜0.05].불동농도신폭로조간AST분비적D-사안산、감안산화γ-안기정산비교차이균유통계학의의(F치분별위26.599、33.539、5.599,P균＜0.05),기중2.5、5.0、10.0 μmol/L신폭로조AST분비적D-사안산[(21.580±1.313)、(21.936±1.539)、(23.401±1.648)μmol/L]、감안산[(26.353±2.449)、(29.711±1.530)、(29.234±2.057)μmol/L]화γ-안기정산[(27.277±3.421)、(30.213±2.098)、(29.364±2.588) μmol/L]균고우0.0 μmol/L신폭로조[(16.017±1.046)、(16.763±3.007)、(22.736±4.139) μmol/L,P균＜0.05].결론 신폭로가인기원대AST분비효질세포원성체질증가,가능회손상학습기억공능.
Objective To investigate the impairment mechanism of learning and memory function induced by arsenite exposure through studying the effects of sodium arsenite on gliotransmitter release from astrocytes.Methods Primary cultured astrocytes were isolated from neonatal (0-3 days) Wistar rats and determined by glial fibrillary acidic protein (GFAP) immunofluorescence staining.The primary cultured astrocytes were randomly divided into four groups,in which astrocytes were exposed to 0.0,2.5,5.0,or 10.0 μmol/L sodium arsenite,respectively,for 24 h.Intracellular free Ca2+ concentration ([Ca2+]i) in astrocytes was measured by fluorescence dual wavelength spectrophotometer;,concentrations of glutamate,D-serine,glycine and γ-aminobutyric acid were measured by high performance liquid chromatography (HPLC).Results More than 95％ cells were positive for GFAP immunofluorescence staining.The difference of [Ca2+]i among groups treated with sodium arsenite was statistically significant (F =20.030,P ＜ 0.05).[Ca2+]i increased significantly in group treated with 10.0 μmol/L sodium arsenite [(263.27 ± 14.80)nmol/L] compared with those in groups treated with 0.0,2.5,5.0 μmol/L sodium arsenite [(204.24 ± 27.21),(214.49 ± 21.85),(232.74 ± 23.14)nmol/L,all P ＜ 0.05].The differences of the levels of D-serine,glycine and γ-aminobutyric acidamong groups treated with sodium arsenite were significant (F =26.599,33.539,5.599,all P ＜ 0.05).The levels of D-serine [(21.580 ± 1.313),(21.936 ± 1.539),(23.401 ± 1.648)μmol/L],glycine [(26.353 ± 2.449),(29.711 ± 1.530),(29.234 ± 2.057)μmol/L] and γ-aminobutyric acid [(27.277 ± 3.421),(30.213 ± 2.098),(29.364 ± 2.588)μmol/L] released by astrocytes increased significantly in groups treated with 2.5,5.0,10.0 μmol/L sodium arsenite compared with those in groups treated with 0.0 μmol/L sodium arsenite [(16.017 ± 1.046),(16.763 ± 3.007),(22.736 ± 4.139)μmol/L,all P ＜ 0.05].Conclusion Arsenite could affect gliotransmitter release from astrocytes,and further impair learning and memory function.
