中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
12期
1303-1307
,共5页
抑制性G蛋白α2亚单位%脑缺血-再灌注损伤%海马%百日咳毒素%凋亡%神经细胞%钙离子%死亡
抑製性G蛋白α2亞單位%腦缺血-再灌註損傷%海馬%百日咳毒素%凋亡%神經細胞%鈣離子%死亡
억제성G단백α2아단위%뇌결혈-재관주손상%해마%백일해독소%조망%신경세포%개리자%사망
Gαi2%Cerebral ischemia reperfusion injury%Hippocampus%Pertussis toxin%Apoptosis%Neuron%Ca2+%Death
目的 探讨分析抑制性G蛋白α2亚单位(Gαi2)在大鼠脑缺血-再灌注损伤模型海马中的表达及其对神经细胞内钙离子浓度和凋亡的影响.方法 将90只SD大鼠随机(随机数字法)分为假手术组(SS组,n=30)、脑缺血-再灌注损伤组(IR组,n=30).脑缺血-再灌注且脑室内微量灌注Gαi2活性抑制剂百日咳毒素(PT)组(PT组,n=30)；用免疫组化、Western blotting分别测定大鼠脑缺血-再灌注后6h、12 h、24 h各组中海马神经内Gαi2的表达,采用流式细胞术测定各组中海马细胞内游离Ca2+浓度的平均荧光值,TUNEL法检测神经元细胞的凋亡.结果 不同时间点,IR组Gαi2含量和Ca2+浓度较SS组明显增高(P＜0.01),PT组Ca2+浓度较IR组升高(P＜0.01),PT组神经细胞凋亡率较IR组升高(P＜0.05).结论 Gαi2在大鼠脑缺血-再灌注损伤中表达明显增高,并可降低缺血神经细胞内的钙离子浓度,减少神经细胞的凋亡,对缺血神经细胞具有保护作用.
目的 探討分析抑製性G蛋白α2亞單位(Gαi2)在大鼠腦缺血-再灌註損傷模型海馬中的錶達及其對神經細胞內鈣離子濃度和凋亡的影響.方法 將90隻SD大鼠隨機(隨機數字法)分為假手術組(SS組,n=30)、腦缺血-再灌註損傷組(IR組,n=30).腦缺血-再灌註且腦室內微量灌註Gαi2活性抑製劑百日咳毒素(PT)組(PT組,n=30)；用免疫組化、Western blotting分彆測定大鼠腦缺血-再灌註後6h、12 h、24 h各組中海馬神經內Gαi2的錶達,採用流式細胞術測定各組中海馬細胞內遊離Ca2+濃度的平均熒光值,TUNEL法檢測神經元細胞的凋亡.結果 不同時間點,IR組Gαi2含量和Ca2+濃度較SS組明顯增高(P＜0.01),PT組Ca2+濃度較IR組升高(P＜0.01),PT組神經細胞凋亡率較IR組升高(P＜0.05).結論 Gαi2在大鼠腦缺血-再灌註損傷中錶達明顯增高,併可降低缺血神經細胞內的鈣離子濃度,減少神經細胞的凋亡,對缺血神經細胞具有保護作用.
목적 탐토분석억제성G단백α2아단위(Gαi2)재대서뇌결혈-재관주손상모형해마중적표체급기대신경세포내개리자농도화조망적영향.방법 장90지SD대서수궤(수궤수자법)분위가수술조(SS조,n=30)、뇌결혈-재관주손상조(IR조,n=30).뇌결혈-재관주차뇌실내미량관주Gαi2활성억제제백일해독소(PT)조(PT조,n=30)；용면역조화、Western blotting분별측정대서뇌결혈-재관주후6h、12 h、24 h각조중해마신경내Gαi2적표체,채용류식세포술측정각조중해마세포내유리Ca2+농도적평균형광치,TUNEL법검측신경원세포적조망.결과 불동시간점,IR조Gαi2함량화Ca2+농도교SS조명현증고(P＜0.01),PT조Ca2+농도교IR조승고(P＜0.01),PT조신경세포조망솔교IR조승고(P＜0.05).결론 Gαi2재대서뇌결혈-재관주손상중표체명현증고,병가강저결혈신경세포내적개리자농도,감소신경세포적조망,대결혈신경세포구유보호작용.
Objective To discuss the levels of inhibitory protein Ga-2 (Gαi2) in hippocampus of brain and the effects of Gαi2 on neuronal intracellular Ca2+ level in cerebral ischemia reperfusion injury model of rats.Methods Ninety SD rats were randomly (random number) assigned to sham group (n =30),ischemic-reperfusion (IR) group (n =30),Pertussis toxin (PT) group (n =30).The blood flow of right common carotid artery of rat was blocked for 90 min to make ischemia reperfusion model.The levels of Gαi2 in hippocampus was assayed by immunohistochemistry and Western blotting after ischemia reperfusion for 6,12,24 h in each group.The average fluorescence values of intracellular Ca2+ levels in hippocampus of rats in three groups were detected by using Flow CytoMeter (FCM).Neuronal cell apoptosis was measured by TUNEL.Results After restoration of middle cerebral artery blood flow for different lengths of time,the levels of hippocampus Gαi2 and Ca2+ levels in IR group were significantly higher than those in Sham group (P ＜0.01).The hippocampus Ca2+ levels in PT group were higher than those in IR group (P ＜ 0.01).The apoptotic rates of neurons in PT group were lower than those in IR group (P ＜0.05).Conclusions The level of Gαi2 in cerebral ischemia reperfusion injury model was increased.Gαi2 might reduce the calcium ion concentration of neurons after cerebral ischemia and rcduce the neuronal cell apoptosis in this model.Gαi2 might play a role in protecting neuron from cerebral ischemia reperfusion injury.
