中华核医学与分子影像杂志
中華覈醫學與分子影像雜誌
중화핵의학여분자영상잡지
Chinese Journal of Nuclear Medicine and Molecular Imaging
2014年
1期
43-47
,共5页
姜春娟%许倩%徐凯%路莉%荣玉涛
薑春娟%許倩%徐凱%路莉%榮玉濤
강춘연%허천%서개%로리%영옥도
脑缺血%再灌注%STAT转录因子%红细胞生成素%磁共振成像%大鼠
腦缺血%再灌註%STAT轉錄因子%紅細胞生成素%磁共振成像%大鼠
뇌결혈%재관주%STAT전록인자%홍세포생성소%자공진성상%대서
Cerebral ischemia%Reperfusion%STAT transcription factors%Erythropoietin%Magnetic resonance imaging%Rats
目的 探讨促红细胞生成素(EPO)对大鼠局灶性脑缺血再灌注后信号转导与转录激活因子(STAT)1、磷酸化STAT1(P-STAT1)、STAT3、P-STAT3蛋白表达及细胞凋亡的影响.方法 将雄性SD大鼠80只完全随机分为假手术(A)组、单纯脑缺血再灌注(B)组、脑缺血再灌注±生理盐水(C)组和脑缺血再灌注±EPO(D)组,每组20只,采用线栓法制作局灶性脑缺血再灌注损伤模型.各组大鼠均于缺血2h后再灌注24 h时,行MRI检查并计算缺血区异常信号相对面积值,Western b1ot检测STAT1、P-STAT1、STAT3、P-STAT3蛋白表达水平的变化并计算相对灰度(rOD)值,原位末端标记法(TUNEL)检测细胞凋亡情况并计算细胞凋亡指数.应用单因素方差分析和q检验分析数据.结果 MRIT2WI示B、C组在左侧大脑半球皮质及皮质下均可见大片脑缺血异常高信号影,D组仅皮质下脑实质内或仅皮质处可见斑片状脑缺血高信号影；与B、C组比较,D组异常高信号区相对面积明显减小[(28.00±4.60)％、(29.70±4.80)％和(21.10±2.40)％；F=11.285,P＜0.01].STAT1、STAT3蛋白在正常脑组织中表达,缺血再灌注及EPO干预对二者表达水平均无明显影响(F=0.806和1.558,均P＞0.05).P-STAT1在A组表达较低,rOD值为0.75±0.13,缺血再灌注后表达显著增加；与B、C组相比,D组P-STAT1表达水平有所减少(B～D:2.08±0.15、2.05±0.16、1.92±0.05；F=3.274,P＞0.05).P-STAT3在A组表达也较低,rOD值为1.02±0.09,缺血再灌注后表达显著增加；与B、C组相比,D组P-STAT3表达明显增加(B～D:2.22±0.13、2.04±0.14、4.21±0.21；F=40.719,P＜0.01).B、C组细胞凋亡指数分别为(42.00±1.30)％和(41.20±2.50)％,高于D组[(20.90±1.46)％；F=378.704,P＜0.01].结论 EPO可增加脑缺血区域P-STAT3表达水平、降低P-STAT1表达水平,从而减少脑细胞凋亡的数目、减小脑梗死面积.
目的 探討促紅細胞生成素(EPO)對大鼠跼竈性腦缺血再灌註後信號轉導與轉錄激活因子(STAT)1、燐痠化STAT1(P-STAT1)、STAT3、P-STAT3蛋白錶達及細胞凋亡的影響.方法 將雄性SD大鼠80隻完全隨機分為假手術(A)組、單純腦缺血再灌註(B)組、腦缺血再灌註±生理鹽水(C)組和腦缺血再灌註±EPO(D)組,每組20隻,採用線栓法製作跼竈性腦缺血再灌註損傷模型.各組大鼠均于缺血2h後再灌註24 h時,行MRI檢查併計算缺血區異常信號相對麵積值,Western b1ot檢測STAT1、P-STAT1、STAT3、P-STAT3蛋白錶達水平的變化併計算相對灰度(rOD)值,原位末耑標記法(TUNEL)檢測細胞凋亡情況併計算細胞凋亡指數.應用單因素方差分析和q檢驗分析數據.結果 MRIT2WI示B、C組在左側大腦半毬皮質及皮質下均可見大片腦缺血異常高信號影,D組僅皮質下腦實質內或僅皮質處可見斑片狀腦缺血高信號影；與B、C組比較,D組異常高信號區相對麵積明顯減小[(28.00±4.60)％、(29.70±4.80)％和(21.10±2.40)％；F=11.285,P＜0.01].STAT1、STAT3蛋白在正常腦組織中錶達,缺血再灌註及EPO榦預對二者錶達水平均無明顯影響(F=0.806和1.558,均P＞0.05).P-STAT1在A組錶達較低,rOD值為0.75±0.13,缺血再灌註後錶達顯著增加；與B、C組相比,D組P-STAT1錶達水平有所減少(B～D:2.08±0.15、2.05±0.16、1.92±0.05；F=3.274,P＞0.05).P-STAT3在A組錶達也較低,rOD值為1.02±0.09,缺血再灌註後錶達顯著增加；與B、C組相比,D組P-STAT3錶達明顯增加(B～D:2.22±0.13、2.04±0.14、4.21±0.21；F=40.719,P＜0.01).B、C組細胞凋亡指數分彆為(42.00±1.30)％和(41.20±2.50)％,高于D組[(20.90±1.46)％；F=378.704,P＜0.01].結論 EPO可增加腦缺血區域P-STAT3錶達水平、降低P-STAT1錶達水平,從而減少腦細胞凋亡的數目、減小腦梗死麵積.
목적 탐토촉홍세포생성소(EPO)대대서국조성뇌결혈재관주후신호전도여전록격활인자(STAT)1、린산화STAT1(P-STAT1)、STAT3、P-STAT3단백표체급세포조망적영향.방법 장웅성SD대서80지완전수궤분위가수술(A)조、단순뇌결혈재관주(B)조、뇌결혈재관주±생리염수(C)조화뇌결혈재관주±EPO(D)조,매조20지,채용선전법제작국조성뇌결혈재관주손상모형.각조대서균우결혈2h후재관주24 h시,행MRI검사병계산결혈구이상신호상대면적치,Western b1ot검측STAT1、P-STAT1、STAT3、P-STAT3단백표체수평적변화병계산상대회도(rOD)치,원위말단표기법(TUNEL)검측세포조망정황병계산세포조망지수.응용단인소방차분석화q검험분석수거.결과 MRIT2WI시B、C조재좌측대뇌반구피질급피질하균가견대편뇌결혈이상고신호영,D조부피질하뇌실질내혹부피질처가견반편상뇌결혈고신호영；여B、C조비교,D조이상고신호구상대면적명현감소[(28.00±4.60)％、(29.70±4.80)％화(21.10±2.40)％；F=11.285,P＜0.01].STAT1、STAT3단백재정상뇌조직중표체,결혈재관주급EPO간예대이자표체수평균무명현영향(F=0.806화1.558,균P＞0.05).P-STAT1재A조표체교저,rOD치위0.75±0.13,결혈재관주후표체현저증가；여B、C조상비,D조P-STAT1표체수평유소감소(B～D:2.08±0.15、2.05±0.16、1.92±0.05；F=3.274,P＞0.05).P-STAT3재A조표체야교저,rOD치위1.02±0.09,결혈재관주후표체현저증가；여B、C조상비,D조P-STAT3표체명현증가(B～D:2.22±0.13、2.04±0.14、4.21±0.21；F=40.719,P＜0.01).B、C조세포조망지수분별위(42.00±1.30)％화(41.20±2.50)％,고우D조[(20.90±1.46)％；F=378.704,P＜0.01].결론 EPO가증가뇌결혈구역P-STAT3표체수평、강저P-STAT1표체수평,종이감소뇌세포조망적수목、감소뇌경사면적.
Objective To explore the effects of erythropoietin (EPO) on the expression of signal transducer and activator of transcription (STAT) 1,phosphorylated STAT1 (P-STAT1),STAT3,P-STAT3 and cell apoptosis in rat models of focal cerebral ischemia-reperfusion.Methods Eighty male SpragueDawley rats were randomly and evenly divided into four groups by completely random design method: shamoperation (group A),cerebral ischemia-reperfusion (group B),cerebral ischemia-reperfusion ± saline (group C) and cerebral ischemia-reperfusion ± EPO (group D).The model of focal cerebral ischemiareperfusion injury was established by blocking the left middle cerebral artery.All rats underwent MRI for the detection of the changes of infarct area between 2 h post ischemia and 24 h of reperfusion.Western blot was used to observe the expression of STAT1,P-STAT1,STAT3,P-STAT3.Terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) was used to evaluate the cell apoptosis including the relative area (ROI area/whole brain area of the same layer × 100％) of abnormal signal region,relative optical density (rOD) and apoptotic index.One-way analysis of variance and q test were used to analyze the data.Results On T2WI imaging,rats in group B and group C presented large hyperintense areas in the cortex and subcortex of left hemispheric ((28.00±4.60)％ and (29.70±4.80)％ respectively).Group D presented less hyperintense areas in the cortex and subcortex of left hemispheric compared with group B and group C ((21.10±2.40) ％; F=11.285,P＜0.01).The expression of STAT1 and STAT3 proteins was not significantly affected by ischemia-reperfusion and EPO intervention compared with normal brain tissue (F=0.806,1.558,both P＞0.05).However,the level of P-STAT1 was low in group A (rOD =0.75±0.13) but increased after cerebral ischemia-reperfusion.Compared with group B and group C,P-STAT1 expression was lower in group D (B-D: 2.08±0.15,2.05±0.16,1.92±0.05; F=3.274,P＞0.05).The level of P-STAT3was also low in group A (rOD=1.02±0.09).Compared with group B and group C,P-STAT3 expression in group D was significandy higher (B-D: 2.22±0.13,2.04±0.14,4.21±0.21 ; F=40.719,P＜0.01).The apoptotic index of group B and group C was (42.00±1.30)％ and (41.20±2.50)％,respectively,which was significantly higher than that of group D ((20.90± 1.46) ％ ; F=378.704,P＜0.01).Conclusion Intraperitoneal injection of EPO can reduce the cerebral ischemic area and the number of apoptotic cells in the ischemic penumbra in rat ischemia-reperfusion models through increasing P-STAT3 and decreasing P-STAT1 levels.
