国际脑血管病杂志
國際腦血管病雜誌
국제뇌혈관병잡지
INTERNATIONAL JOURNAL OF CEREBROVASCULAR DISEASES
2013年
3期
181-185
,共5页
肖国栋%成松明%孙涛%曹勇军
肖國棟%成鬆明%孫濤%曹勇軍
초국동%성송명%손도%조용군
核受体亚家族4,A组,成员1%神经元%小脑%细胞,培养的%脑缺血%氧%葡萄糖%细胞凋亡%胱天蛋白酶3%细胞色素c类%神经保护药%大鼠
覈受體亞傢族4,A組,成員1%神經元%小腦%細胞,培養的%腦缺血%氧%葡萄糖%細胞凋亡%胱天蛋白酶3%細胞色素c類%神經保護藥%大鼠
핵수체아가족4,A조,성원1%신경원%소뇌%세포,배양적%뇌결혈%양%포도당%세포조망%광천단백매3%세포색소c류%신경보호약%대서
Nuclear Receptor Subfamily 4,Group A,Member 1%Neurons%Cerebellum%Cells,Cultured%Brain Ischemia%Oxygen%Glucose%Apoptosis%Caspase 3%Cytochromes c%Neuroprotective Agents%Rats
目的 探讨孤儿核受体NR4A1表达对氧-葡萄糖剥夺(oxy gen-glucose deprivation,OGD)诱导培养大鼠小脑颗粒神经元凋亡的影响及其可能机制.方法 大鼠小脑颗粒细胞原代培养7～8 d后给予OGD处理.应用四甲基偶氮唑蓝法检测细胞活性,流式细胞术检测细胞凋亡率,蛋白质印迹法检测NR4A1、胱天蛋白酶-3和细胞色素c蛋白表达,实时定量聚合酶链反应检测NR4A1mRNA表达.用编码NR4A1的慢病毒载体转染大鼠小脑颗粒神经元,并检测转染大鼠小脑颗粒神经元OGD后凋亡率以及细胞色素c和胱天蛋白酶-3表达.结果 随着OGD时间的延长,培养大鼠小脑颗粒神经元活性显著性降低,凋亡率显著性增高,NR4A1 mRNA和蛋白以及胱天蛋白酶-3和细胞色素c蛋白表达均显著性上调.转染NR4A1的大鼠小脑颗粒神经元过表达NR4A1,OGD后凋亡率显著性降低,胱天蛋白酶-3和细胞色素c蛋白表达显著性下调.结论 NR4A1过表达能通过下调胱天蛋白酶-3和细胞色素c表达减少OGD诱导的大鼠小脑颗粒神经元凋亡.
目的 探討孤兒覈受體NR4A1錶達對氧-葡萄糖剝奪(oxy gen-glucose deprivation,OGD)誘導培養大鼠小腦顆粒神經元凋亡的影響及其可能機製.方法 大鼠小腦顆粒細胞原代培養7～8 d後給予OGD處理.應用四甲基偶氮唑藍法檢測細胞活性,流式細胞術檢測細胞凋亡率,蛋白質印跡法檢測NR4A1、胱天蛋白酶-3和細胞色素c蛋白錶達,實時定量聚閤酶鏈反應檢測NR4A1mRNA錶達.用編碼NR4A1的慢病毒載體轉染大鼠小腦顆粒神經元,併檢測轉染大鼠小腦顆粒神經元OGD後凋亡率以及細胞色素c和胱天蛋白酶-3錶達.結果 隨著OGD時間的延長,培養大鼠小腦顆粒神經元活性顯著性降低,凋亡率顯著性增高,NR4A1 mRNA和蛋白以及胱天蛋白酶-3和細胞色素c蛋白錶達均顯著性上調.轉染NR4A1的大鼠小腦顆粒神經元過錶達NR4A1,OGD後凋亡率顯著性降低,胱天蛋白酶-3和細胞色素c蛋白錶達顯著性下調.結論 NR4A1過錶達能通過下調胱天蛋白酶-3和細胞色素c錶達減少OGD誘導的大鼠小腦顆粒神經元凋亡.
목적 탐토고인핵수체NR4A1표체대양-포도당박탈(oxy gen-glucose deprivation,OGD)유도배양대서소뇌과립신경원조망적영향급기가능궤제.방법 대서소뇌과립세포원대배양7～8 d후급여OGD처리.응용사갑기우담서람법검측세포활성,류식세포술검측세포조망솔,단백질인적법검측NR4A1、광천단백매-3화세포색소c단백표체,실시정량취합매련반응검측NR4A1mRNA표체.용편마NR4A1적만병독재체전염대서소뇌과립신경원,병검측전염대서소뇌과립신경원OGD후조망솔이급세포색소c화광천단백매-3표체.결과 수착OGD시간적연장,배양대서소뇌과립신경원활성현저성강저,조망솔현저성증고,NR4A1 mRNA화단백이급광천단백매-3화세포색소c단백표체균현저성상조.전염NR4A1적대서소뇌과립신경원과표체NR4A1,OGD후조망솔현저성강저,광천단백매-3화세포색소c단백표체현저성하조.결론 NR4A1과표체능통과하조광천단백매-3화세포색소c표체감소OGD유도적대서소뇌과립신경원조망.
Objective To investigate the effect of orphan nuclear receptor NR4A1 expression on oxygenglucose deprivation (OGD)-induced apoptosis in cultured rat cerebellar granule neurons and its possible mechanisms.Methods Primary rat cerebellar granule neurons were cultured for 7 to 8 days,and then treated with OGD.The activity of cultured rat cerebellar granule neurons was assessed by methyl thiazolyl tetrazolium (MTT) assay,apoptosis was detected with flow cytometry,the expressions of NR4A1,caspase-3 and cytochrome c were determined by Western blot analysis,NR4A1 mRNA expression was detected with real-time polymerase chain reaction.The rat cerebellar granule neurons were transfected with lentiviral vector-encoding rat NR4A1.The apoptotic rates and expressions of caspase-3 and cytochrome c in rat cerebellar granule neurons transfected with NR4A1 were detected after OGD.Results The activity of rat cerebellar granule neurons decreased significantly,the apoptotic rate increased significantly,the expressions of NR4A1 mRNA and protein as well as caspase-3 and cytochrome c incrased significantly along with the OGD time.NR4A1 was overexpressed,apoptosis rate was significantly reduced in rat cerebellar granule neurons transfected with NR4A1.The expressions of caspase-3 and cytochrome c were significantly reduced in the rat cerebellar granule neurons transfected with NR4A1 after OGD.Conclusions NR4A1 overexpression may reduce OGD-induced apoptosis in rat cerebellar granule neurons by downregulating the expressions of caspase-3 and cytochrome c.
