中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2015年
2期
230-233
,共4页
李艳丽%张山%敖利%贾建丽%边庆虎%杨丽娜
李豔麗%張山%敖利%賈建麗%邊慶虎%楊麗娜
리염려%장산%오리%가건려%변경호%양려나
磷脂酰肌醇-3-激酶%蛋白质丝氨酸苏氨酸激酶%低温,人工%再灌注损伤%脑
燐脂酰肌醇-3-激酶%蛋白質絲氨痠囌氨痠激酶%低溫,人工%再灌註損傷%腦
린지선기순-3-격매%단백질사안산소안산격매%저온,인공%재관주손상%뇌
Phosphoinositide-3-kinase%Protein-serine-threonine kinases%Hypothermia,induced%Reperfusion injury%Brain
目的 评价磷脂酰肌醇3-激酶-丝氨酸-苏氨酸蛋白激酶(PI3K/Akt)信号通路在头部浅低温减轻大鼠全脑缺血再灌注损伤中的作用.方法 健康雄性SD大鼠60只,体重250~ 280 g,采用随机数字表法分为5组(n=12):假手术组(S组)、脑缺血再灌注组(I/R组)、浅低温组(H组)、浅低温+溶剂对照组(DM组)和浅低温+PI3K抑制剂LY294002组(LY组).除S组外,其余各组采用改良Pulsinelli四血管阻断法建立全脑缺血再灌注损伤模型,H组采用鼻咽腔降温法降低海马温度至33℃时夹闭双侧颈总动脉15 min复灌,期间维持海马温度32.5~ 33.5℃1h,DM组和LY组于左侧脑室缓慢注射DMSO 5μl或LY294002 5μl,20 min后处理同H组.于再灌注8h时,每组随机取6只大鼠,采用免疫组化法测定海马CA1区磷酸化FoxO3a(pFoxO3a)、Bcl-2及Bax蛋白表达水平,计算Bcl/Bax比值;余6只大鼠,采用Western blot法检测海马磷酸化Akt(p-Akt)表达水平.结果 与S组比较,I/R组、H组和DM组p-Akt、pFoxO3a、Bax蛋白表达上调,H组和DM组Bcl-2蛋白表达下调、Bcl-2/Bax比值升高,I/R组和LY组Bcl-2/Bax比值降低(P<0.05);与I/R组比较,H组和DM组p-Akt、pFoxO3a、Bcl-2蛋白表达上调,Bax蛋白表达下调,Bcl-2/Bax比值升高(P<0.05);与H组和DM组比较,LY组p-Akt、pFoxO3a和Bcl-2蛋白表达下调,Bax蛋白表达上调,Bcl-2/Bax比值降低(P<0.05).结论 PI3K/Akt信号通路参与了头部浅低温减轻大鼠全脑缺血再灌注损伤的过程.
目的 評價燐脂酰肌醇3-激酶-絲氨痠-囌氨痠蛋白激酶(PI3K/Akt)信號通路在頭部淺低溫減輕大鼠全腦缺血再灌註損傷中的作用.方法 健康雄性SD大鼠60隻,體重250~ 280 g,採用隨機數字錶法分為5組(n=12):假手術組(S組)、腦缺血再灌註組(I/R組)、淺低溫組(H組)、淺低溫+溶劑對照組(DM組)和淺低溫+PI3K抑製劑LY294002組(LY組).除S組外,其餘各組採用改良Pulsinelli四血管阻斷法建立全腦缺血再灌註損傷模型,H組採用鼻嚥腔降溫法降低海馬溫度至33℃時夾閉雙側頸總動脈15 min複灌,期間維持海馬溫度32.5~ 33.5℃1h,DM組和LY組于左側腦室緩慢註射DMSO 5μl或LY294002 5μl,20 min後處理同H組.于再灌註8h時,每組隨機取6隻大鼠,採用免疫組化法測定海馬CA1區燐痠化FoxO3a(pFoxO3a)、Bcl-2及Bax蛋白錶達水平,計算Bcl/Bax比值;餘6隻大鼠,採用Western blot法檢測海馬燐痠化Akt(p-Akt)錶達水平.結果 與S組比較,I/R組、H組和DM組p-Akt、pFoxO3a、Bax蛋白錶達上調,H組和DM組Bcl-2蛋白錶達下調、Bcl-2/Bax比值升高,I/R組和LY組Bcl-2/Bax比值降低(P<0.05);與I/R組比較,H組和DM組p-Akt、pFoxO3a、Bcl-2蛋白錶達上調,Bax蛋白錶達下調,Bcl-2/Bax比值升高(P<0.05);與H組和DM組比較,LY組p-Akt、pFoxO3a和Bcl-2蛋白錶達下調,Bax蛋白錶達上調,Bcl-2/Bax比值降低(P<0.05).結論 PI3K/Akt信號通路參與瞭頭部淺低溫減輕大鼠全腦缺血再灌註損傷的過程.
목적 평개린지선기순3-격매-사안산-소안산단백격매(PI3K/Akt)신호통로재두부천저온감경대서전뇌결혈재관주손상중적작용.방법 건강웅성SD대서60지,체중250~ 280 g,채용수궤수자표법분위5조(n=12):가수술조(S조)、뇌결혈재관주조(I/R조)、천저온조(H조)、천저온+용제대조조(DM조)화천저온+PI3K억제제LY294002조(LY조).제S조외,기여각조채용개량Pulsinelli사혈관조단법건립전뇌결혈재관주손상모형,H조채용비인강강온법강저해마온도지33℃시협폐쌍측경총동맥15 min복관,기간유지해마온도32.5~ 33.5℃1h,DM조화LY조우좌측뇌실완만주사DMSO 5μl혹LY294002 5μl,20 min후처리동H조.우재관주8h시,매조수궤취6지대서,채용면역조화법측정해마CA1구린산화FoxO3a(pFoxO3a)、Bcl-2급Bax단백표체수평,계산Bcl/Bax비치;여6지대서,채용Western blot법검측해마린산화Akt(p-Akt)표체수평.결과 여S조비교,I/R조、H조화DM조p-Akt、pFoxO3a、Bax단백표체상조,H조화DM조Bcl-2단백표체하조、Bcl-2/Bax비치승고,I/R조화LY조Bcl-2/Bax비치강저(P<0.05);여I/R조비교,H조화DM조p-Akt、pFoxO3a、Bcl-2단백표체상조,Bax단백표체하조,Bcl-2/Bax비치승고(P<0.05);여H조화DM조비교,LY조p-Akt、pFoxO3a화Bcl-2단백표체하조,Bax단백표체상조,Bcl-2/Bax비치강저(P<0.05).결론 PI3K/Akt신호통로삼여료두부천저온감경대서전뇌결혈재관주손상적과정.
Objective To evaluate the role of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway in mild head hypothermia-induced reduction of global cerebral ischemiareperfusion (I/R) injury in rats.Methods Sixty male Sprague-Dawley rats,weighing 250-280 g,were randomly divided into 5 groups (n=12 each) using a random number table:sham operation group (group S),group I/R,mild hypothermia + I/R group (group H),mild hypothermia + I/R + solvent control group (group DM),and mild hypothermia + I/R + PI3K inhibitor LY294002 group (group LY).Global cerebral I/R was induced by modified four-vessel occlusion method described by Pulsinelli.In H group,when the hippocampal temperature was decreased to 33℃ using nasopharyngeal cooling,the bilateral common carotid arteries were occluded for 15 min followed by reperfusion,and hippocampal hypothermia was maintained at 32.5-33.5 ℃ for 1 h.In DM and LY groups,DMSO and LY294002 5 μl were injected into the left ventricle,respectively,and 20 min later the other procedures were similar to those previously described in group H.At 8 h of reperfusion,6 rats were sacrificed,and hippocampal specimens were obtained to detect the expression of pFoxO3a,Bcl-2 and Bax (by immuno-histochemistry).The expression of phosphor-Akt (p-Akt) was determined by Western blot.Results Compared with group S,the expression of p-Akt,pFoxO3a and Bcl-2 was significantly up-regulated in I/R,H and DM groups,the expression of Bcl-2 was down-regulated,and the ratio of Bcl-2/Bax was increased in H and DM groups,and the ratio of Bcl-2/Bax was decreased in I/R and LY groups.Compared with group I/R,the expression of p-Akt,pFoxO3a and Bcl-2 was significantly up-regulated,the expression of Bcl-2 was down-regulated,and the ratio of Bcl-2/ Bax was increased in H and DM groups.Compared with H and DM groups,the expression of p-Akt,pFoxO3a and Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the ratio of Bcl-2/ Bax was decreased in group LY.Conclusion PI3K/Akt signaling pathway is involved in reduction of global cerebral I/R injury by mild head hypothermia in rats.