神经科学通报(英文版)
神經科學通報(英文版)
신경과학통보(영문판)
NEUROSCIENCE BULLETIN
2008年
4期
231-243
,共13页
孙景军%Ying LIU%叶诸榕
孫景軍%Ying LIU%葉諸榕
손경군%Ying LIU%협제용
P2Y1受体%胶质化%胶质原纤维酸性蛋白%胶质细胞源性神经营养因子%PI3-K/Akt/CREB%JAK2/STAT3%Ras/ERK
P2Y1受體%膠質化%膠質原纖維痠性蛋白%膠質細胞源性神經營養因子%PI3-K/Akt/CREB%JAK2/STAT3%Ras/ERK
P2Y1수체%효질화%효질원섬유산성단백%효질세포원성신경영양인자%PI3-K/Akt/CREB%JAK2/STAT3%Ras/ERK
P2Y1 receptor%gliosis%glial fibriUary acidic protein%glial cell line-derived neurotrophic factor%PI3-K/Akt/CREB%JAK2/STAT3%Ras/ERK
目的 研究P2Y1受体对缺血时星形胶质细胞产生胶质原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)的影响及其相关信号通路.方法 分别利用右侧大脑中动脉线拴阻塞及培养细胞缺氧无营养后恢复正常培养,造成体内、外缺血再灌注模型.用免疫荧光标记、实时定量RT-PCR、Western blotting、酶联免疫吸附试验观察P2Y1受体、GDNF定位,检测GFAP、GDNF及信号分子的表达变化.结果 与单纯性缺血组比较,用选择性拮抗剂MRS2179阻断P2Y1受体后,可使体内、外星形胶质细胞产生的GFAP减少,同时使其产生GDNF增加.体外缺氧无营养并阻断P2Y1受体后:可使磷酸化蛋白激酶B(Akt)及cAMP反应元件结合蛋白(cAMP response element binding protein,CREB)升高,而使磷酸化JAK2及STAT3(Ser727)降低;JAK2的抑制剂AG490在降低磷酸化STAT3(Ser727)的同时也降低GFAP表达水平;P13-K的抑制剂LY294002可降低磷酸化的Akt及CREB;MEK1/2抑制剂U0126可同时降低磷酸化的JAK2、STAT3(Ser727)、Akt及CREB.结论 P2Y1受体参与短时性缺血时星形胶质细胞GFAP及GDNF的产生过程,相关信号途径分别为JAK2/STAT3和P13-K/AKT/CREB,并且两条途径存在串话.
目的 研究P2Y1受體對缺血時星形膠質細胞產生膠質原纖維痠性蛋白(glial fibrillary acidic protein,GFAP)及膠質細胞源性神經營養因子(glial cell line-derived neurotrophic factor,GDNF)的影響及其相關信號通路.方法 分彆利用右側大腦中動脈線拴阻塞及培養細胞缺氧無營養後恢複正常培養,造成體內、外缺血再灌註模型.用免疫熒光標記、實時定量RT-PCR、Western blotting、酶聯免疫吸附試驗觀察P2Y1受體、GDNF定位,檢測GFAP、GDNF及信號分子的錶達變化.結果 與單純性缺血組比較,用選擇性拮抗劑MRS2179阻斷P2Y1受體後,可使體內、外星形膠質細胞產生的GFAP減少,同時使其產生GDNF增加.體外缺氧無營養併阻斷P2Y1受體後:可使燐痠化蛋白激酶B(Akt)及cAMP反應元件結閤蛋白(cAMP response element binding protein,CREB)升高,而使燐痠化JAK2及STAT3(Ser727)降低;JAK2的抑製劑AG490在降低燐痠化STAT3(Ser727)的同時也降低GFAP錶達水平;P13-K的抑製劑LY294002可降低燐痠化的Akt及CREB;MEK1/2抑製劑U0126可同時降低燐痠化的JAK2、STAT3(Ser727)、Akt及CREB.結論 P2Y1受體參與短時性缺血時星形膠質細胞GFAP及GDNF的產生過程,相關信號途徑分彆為JAK2/STAT3和P13-K/AKT/CREB,併且兩條途徑存在串話.
목적 연구P2Y1수체대결혈시성형효질세포산생효질원섬유산성단백(glial fibrillary acidic protein,GFAP)급효질세포원성신경영양인자(glial cell line-derived neurotrophic factor,GDNF)적영향급기상관신호통로.방법 분별이용우측대뇌중동맥선전조새급배양세포결양무영양후회복정상배양,조성체내、외결혈재관주모형.용면역형광표기、실시정량RT-PCR、Western blotting、매련면역흡부시험관찰P2Y1수체、GDNF정위,검측GFAP、GDNF급신호분자적표체변화.결과 여단순성결혈조비교,용선택성길항제MRS2179조단P2Y1수체후,가사체내、외성형효질세포산생적GFAP감소,동시사기산생GDNF증가.체외결양무영양병조단P2Y1수체후:가사린산화단백격매B(Akt)급cAMP반응원건결합단백(cAMP response element binding protein,CREB)승고,이사린산화JAK2급STAT3(Ser727)강저;JAK2적억제제AG490재강저린산화STAT3(Ser727)적동시야강저GFAP표체수평;P13-K적억제제LY294002가강저린산화적Akt급CREB;MEK1/2억제제U0126가동시강저린산화적JAK2、STAT3(Ser727)、Akt급CREB.결론 P2Y1수체삼여단시성결혈시성형효질세포GFAP급GDNF적산생과정,상관신호도경분별위JAK2/STAT3화P13-K/AKT/CREB,병차량조도경존재천화.
Objective The present study aimed to explore the role of P2Y1 receptor in glial fibrillary acidic protein (GFAP) production and glial cell line-derived neurotrophic factor (GDNF) secretion of astrocytes under ischemic insult and the related signaling pathways. Methods Using transient right middle cerebral artery occlusion (tMCAO) and oxygen-glucose-serum deprivation for 2 h as the model of ischemic injury in vivo and in vitro, immunofluorescence, quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, enzyme linked immunosorbent assay (ELISA) were used to investigate location of P2Y1 receptor and GDNF, the expression of GFAP and GDNF, and the changes of signaling molecules. Results Blockage of P2Y1 receptor with the selective antagonist N6-methyl-2' -deoxyadenosine 3',5'-bisphosphate diammonium (MRS2179) reduced GFAP production and increased GDNF production in the antagonist group as compared with simple iscbemic group both in vivo and in vitro. Oxygen-glucose-serum deprivation and blockage of P2Y1 receptor caused elevation of phosphorylated Akt and cAMP response element binding protein (CREB), and reduction of phosphorylated Janus kinase2 (JAK2) and signal transducer and activator of transeription3 (STAT3, Ser727). After blockage of P2Y1 receptor and deprivation of oxygen-glucose-serum, AG490 (inhibitor of JAK2) reduced phosphorylation of STAT3 (Ser727) as well as expression of GFAP; LY294002, an inhibitor of phosphatidylinositol 3-kinase (PI3-K), decreased phosphorylation of Akt and CREB; the inhibitor of mitogen-activated protein kinase kinasel/2 (MEK1/2) U0126, an important molecule of Ras/extracellular signal- regulated kinase (ERK) signaling pathway, decreased the phosphorylafion of JAK2, STAT3 (Ser727), Akt and CREB. Conclusion These results suggest that P2Y1 receptor plays a role in the production of GFAP and GDNF in astrocytes under transient ischemic condition and the related signaling pathways may be JAK2/STAT3 and PI3-K/Akt/CREB, respectively, and that crosstalk probably exists between them.
