CAJ | 학술논문

目的观察尼古丁慢性摄入加重大鼠脑缺血再灌注损伤性脑水肿的程度,并初步探讨其作用机制.方法 10周龄雄性Sprague Dawley(SD)大鼠90只,按数字表法随机分为对照组(45只)和尼古丁组(45只),分别给予生理盐水[0.5 ml/(kg·d)]、尼古丁[3 mg/(kg·d)]皮下注射,1次/d.6周后将2组大鼠制作成大脑中动脉缺血再灌注损伤模型,分5个时间点(每组每个时间点9只)观察脑水肿的发生情况和程度.取对照组和尼古丁组脑微血管内皮细胞株bEnd.3进行培养,分为3组:对照组、尼古丁组(1μmol/L)、尼古丁+α-金环蛇毒素组(尼古丁1 μmol/L,α-金环蛇毒素0.1μmol/L),检测3组细胞通透性的变化及细胞中水通道蛋白1(aquaporin 1,AQP1)的表达变化.结果与对照组比较,尼古丁组再灌注3.0h后脑水肿的程度明显加重,再灌注12.0 h时脑组织含水量最大[2组分别为(86.53±2.26)％和(90.47±1.94)％],差异有统计学意义(P＜0.01).对脑微血管内皮细胞株bEnd.3的检测发现,尼古丁组细胞通透性[(143±11)％]明显高于对照组[(100±13)％],差异有统计学意义(P＜0.01);尼古丁+α-金环蛇毒素组通透性[(114±15)％]较尼古丁组明显改善,差异有统计学意义(P＜0.05).细胞内AQP1蛋白的表达,尼古丁组(1.49±0.19)明显高于对照组(1.04±0.12),差异有统计学意义(P＜0.01);尼古丁+α-金环蛇毒素组通透性(1.20 ±0.14)较尼古丁组有所减少,差异有统计学意义(P＜0.05).结论在尼古丁刺激下,脑微血管内皮细胞通透性的增加是脑水肿加重的原因.内皮细胞中α7尼古丁乙酰胆碱受体(α7nAchR)和AQP1蛋白参与了其中的发生过程.
목적 관찰니고정만성섭입가중대서뇌결혈재관주손상성뇌수종적정도,병초보탐토기작용궤제.방법 10주령웅성Sprague Dawley(SD)대서90지,안수자표법수궤분위대조조(45지)화니고정조(45지),분별급여생리염수[0.5 ml/(kg·d)]、니고정[3 mg/(kg·d)]피하주사,1차/d.6주후장2조대서제작성대뇌중동맥결혈재관주손상모형,분5개시간점(매조매개시간점9지)관찰뇌수종적발생정황화정도.취대조조화니고정조뇌미혈관내피세포주bEnd.3진행배양,분위3조:대조조、니고정조(1μmol/L)、니고정+α-금배사독소조(니고정1 μmol/L,α-금배사독소0.1μmol/L),검측3조세포통투성적변화급세포중수통도단백1(aquaporin 1,AQP1)적표체변화.결과 여대조조비교,니고정조재관주3.0h후뇌수종적정도명현가중,재관주12.0 h시뇌조직함수량최대[2조분별위(86.53±2.26)％화(90.47±1.94)％],차이유통계학의의(P＜0.01).대뇌미혈관내피세포주bEnd.3적검측발현,니고정조세포통투성[(143±11)％]명현고우대조조[(100±13)％],차이유통계학의의(P＜0.01);니고정+α-금배사독소조통투성[(114±15)％]교니고정조명현개선,차이유통계학의의(P＜0.05).세포내AQP1단백적표체,니고정조(1.49±0.19)명현고우대조조(1.04±0.12),차이유통계학의의(P＜0.01);니고정+α-금배사독소조통투성(1.20 ±0.14)교니고정조유소감소,차이유통계학의의(P＜0.05).결론 재니고정자격하,뇌미혈관내피세포통투성적증가시뇌수종가중적원인.내피세포중α7니고정을선담감수체(α7nAchR)화AQP1단백삼여료기중적발생과정.
Objective To observe the effect of chronic nicotine intake on cerebral edema induced by ischemic reperfusion in rats and also to investigate the mechanism involved.Methods Ninety male SD rats,with an age of 10 weeks,were randomly divided into the normal saline group (n =45),and the nicotine group (n =45).The animals in the 2 groups were respectively given physiological saline (0.5 ml/kg · d) and nicotine (subcutaneous injection with nicotine at a dosage of 3mg/kg · d) for 6 weeks.After 6 weeks,models of cerebral injury induced by ischemic reperfusion were developed,and occurrence and seriousness of cerebral edema were observed at 5 different time points (there were 9 animals in each group at each different time point).Endothelial cell strains from cerebral micro vessels,bEnd.3,were collected for culture from the animals of the control group and the nicotine group.The animals in the nicotine group were further divided into zthe nicotine group (1μmol/L) and the nicotine + α-bungarotoxin group (nicotine 1 μmol/L,α-bungarotoxin 0.1μmol/L).Changes in cell permeability and the expressions of aquaporin 1 (AQP1) were detected in the animals of the 3 groups.Results Three hours after ischemic reperfusion,cerebral edema in the animals of the nicotine group obviously worsened,as compared with that of the animals in the control group.Cerebral water contents of the 2 groups came to peak levels at 12.0 h after ischemic reperfusion,reaching the levels of(86.53 ± 2.26) ％ and(90.47 ± 1.94) ％ respectively,with statistical significance (P ＜ 0.01).Detection of bEnd.3 in the cerebral micro-vessel endothelial cell strains revealed that cell permeability in the nicotine group [(114 ± 15)％] was significantly higher than that of the control group [(100 ± 13)％],with statistical significance (P ＜0.01).Cell permeability in the nicotine + α-bungarotoxin group significantly improved,as compared with that of the nicotine group,also with statistical significance(P ＜ 0.05).The expression of AQP1 in the nicotine group (1.49 ± 0.19) was significantly higher than that of the control group (1.04 ± 0.12),with statistical significance(P ＜0.01).Cell permeability in the nicotine + α-bungarotoxin group(1.20 ±0.14)was slightly lower than that of the nicotine group,also with statistical significance(P ＜ 0.05).Conclusions With the stimulation of nicotine,increase in the cerebral endothelial cell permeability was responsible for the exacerbation of cerebral edema.The α7nAchR and AQP1 in endothelial cells might be involved in the whole process.