中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2014年
6期
647-651
,共5页
重症中暑%脑损害%热打击%热打击模型%神经元细胞%凋亡%凋亡机制%活性氧
重癥中暑%腦損害%熱打擊%熱打擊模型%神經元細胞%凋亡%凋亡機製%活性氧
중증중서%뇌손해%열타격%열타격모형%신경원세포%조망%조망궤제%활성양
Severe heat stroke%Brain damage%Heat stress%Heat stress model%Neuron%Apoptosis%Apoptosismechanism%Reactive oxygen species
目的：通过观察热打击诱导活性氧（reactive oxygen species，ROS）爆发性增多对神经元细胞凋亡的影响，探讨重症中暑所致脑损害的发病机理。方法建立神经元细胞热打击模型，对照组将细胞置于标准37℃、5％CO2细胞培养箱，热打击组将细胞置于43℃细胞培养箱中热打击2 h，热打击后继续37℃、5％CO2细胞培养箱孵育，DCFH法检测热打击后0 h、0.5 h、1 h、2 h细胞内ROS含量；Annexin V-FITC/PI双染色方法和Westen blot检测热打击后0 h、3 h、6 h、12 h细胞凋亡率及caspase-3蛋白表达，同时检测ROS特异性清除剂MnTMPyP对热打击后12 h细胞凋亡的影响。结果与对照组比较，43℃热打击后0 h细胞内ROS增加，2 hROS呈爆发性增多（P＜0.05）；43℃热打击后3 h神经元细胞开始出现凋亡，12 h 诱导大量凋亡（43.2％，P＜0.05）、caspase-3蛋白表达明显增加（P＜0.05），其趋势与细胞凋亡趋势一致；MnTMPyP明显抑制了43℃热打击后12 h神经元细胞凋亡率（47.42％降至18.45％，P＜0.05）及caspase-3蛋白的表达。结论 ROS作为上游信号分子介导了热打击诱导的神经元细胞凋亡，但其介导神经元细胞凋亡的中间机制尚需进一步研究。
目的：通過觀察熱打擊誘導活性氧（reactive oxygen species，ROS）爆髮性增多對神經元細胞凋亡的影響，探討重癥中暑所緻腦損害的髮病機理。方法建立神經元細胞熱打擊模型，對照組將細胞置于標準37℃、5％CO2細胞培養箱，熱打擊組將細胞置于43℃細胞培養箱中熱打擊2 h，熱打擊後繼續37℃、5％CO2細胞培養箱孵育，DCFH法檢測熱打擊後0 h、0.5 h、1 h、2 h細胞內ROS含量；Annexin V-FITC/PI雙染色方法和Westen blot檢測熱打擊後0 h、3 h、6 h、12 h細胞凋亡率及caspase-3蛋白錶達，同時檢測ROS特異性清除劑MnTMPyP對熱打擊後12 h細胞凋亡的影響。結果與對照組比較，43℃熱打擊後0 h細胞內ROS增加，2 hROS呈爆髮性增多（P＜0.05）；43℃熱打擊後3 h神經元細胞開始齣現凋亡，12 h 誘導大量凋亡（43.2％，P＜0.05）、caspase-3蛋白錶達明顯增加（P＜0.05），其趨勢與細胞凋亡趨勢一緻；MnTMPyP明顯抑製瞭43℃熱打擊後12 h神經元細胞凋亡率（47.42％降至18.45％，P＜0.05）及caspase-3蛋白的錶達。結論 ROS作為上遊信號分子介導瞭熱打擊誘導的神經元細胞凋亡，但其介導神經元細胞凋亡的中間機製尚需進一步研究。
목적：통과관찰열타격유도활성양（reactive oxygen species，ROS）폭발성증다대신경원세포조망적영향，탐토중증중서소치뇌손해적발병궤리。방법건립신경원세포열타격모형，대조조장세포치우표준37℃、5％CO2세포배양상，열타격조장세포치우43℃세포배양상중열타격2 h，열타격후계속37℃、5％CO2세포배양상부육，DCFH법검측열타격후0 h、0.5 h、1 h、2 h세포내ROS함량；Annexin V-FITC/PI쌍염색방법화Westen blot검측열타격후0 h、3 h、6 h、12 h세포조망솔급caspase-3단백표체，동시검측ROS특이성청제제MnTMPyP대열타격후12 h세포조망적영향。결과여대조조비교，43℃열타격후0 h세포내ROS증가，2 hROS정폭발성증다（P＜0.05）；43℃열타격후3 h신경원세포개시출현조망，12 h 유도대량조망（43.2％，P＜0.05）、caspase-3단백표체명현증가（P＜0.05），기추세여세포조망추세일치；MnTMPyP명현억제료43℃열타격후12 h신경원세포조망솔（47.42％강지18.45％，P＜0.05）급caspase-3단백적표체。결론 ROS작위상유신호분자개도료열타격유도적신경원세포조망，단기개도신경원세포조망적중간궤제상수진일보연구。
Objective To observe the effect of heat stress-induced burst out of reactive oxygen on neuronal apoptosis and investigate pathogenesis of brain damage caused by severe heat stroke.Methods Neurons heat stress model is set up.Control group were incubated at 37 ℃,5%CO2 ,While heat stress group of cells were incubated at 43 ℃for 2 h,then all the cells were further incubated at 37 ℃for different time as indicated.The amounts of ROS were assayed by DCFH staining at 0 h,0.5 h,1 h,2 h after heat stress.Apoptosis was analyzed by flow cytometry using Annexin V-FITC/PI staining and expression of caspase-3 were determined by westen blotat 0 h、3 h、6 h、12 h after heat stress.In addition,MnTMPyP is the specificscavengers of ROS,which effect on apoptosis is also studied at 12 h after heat stress.Results Compared with control group,amounts of ROS was significant increased at 0 h after heat stress,the burst out of it was at 2 h after heat stress (P<0.05 ).Apoptosis was induced at 3h after heat stress ,it was significant increased at 12 h after heat stress (43.2%,P<0.05 ).The expression of caspase-3 was also significant increased at 12 h after heat stress (P<0.05 ),and its trend was consistent with apoptosis rate trend.In addition,the scavengers MnTMPyP significantly decreased the apoptosis (47.42% to 18.45%, P<0.05 )and expression of caspase-3 at 12 h after heat stress.Conclusions An upstream signaling molecules,ROS could mediate heat stress-induced neuronal apoptosis,but its intermediate mechanism needs for further studies.
