摘要
目的 探讨褪黑素对大鼠脑缺血-再灌注损伤(CIRI)后大脑皮质细胞毒性水肿的影响及其机制。方法 将60只成年健康无特定病原体雄性Sprague Dawley大鼠完全随机分为假手术组(Sham组,12只)、CIRI组24只、褪黑素治疗组(24只)。采用改良的Zea-Longa线栓法建立大鼠CIRI模型,其中褪黑素治疗组在建模前、后30 min腹腔注射褪黑素(5 mg/kg);CIRI组在建模前、后30 min经腹腔注射等量的等渗盐水。Sham组分离右侧颈总动脉后缝合皮肤,并于假手术前、后30 min注射等量的等渗盐水。建模后2、6、12、24 h及72 h, 3组大鼠(每组6只)均行活体MRI T2加权成像(T2WI)、扩散加权成像(DWI)检查,结合表观扩散系数(ADC)图,动态分析褪黑素干预后CIRI大鼠脑梗死体积百分比、脑水肿百分比及大脑皮质相对ADC(rADC)值的变化。建模后6、24 h及72 h,采用脱氧核苷酸末端转移酶介导的2′-脱氧尿苷5′-三磷酸盐介导的缺口末端标记(TUNEL)法观察褪黑素对CIRI大鼠缺血侧大脑皮质细胞凋亡的影响;免疫荧光双标法观察褪黑素干预对星形胶质细胞水通道蛋白4(AQP4)和钠-钾-氯协同转运蛋白1(NKCC1)的影响;免疫荧光染色结合蛋白质印记,探讨褪黑素对离子钙接头蛋白抗原(Iba-1)标记的小胶质细胞活化的影响。建模后24 h,苏木素-伊红(HE)染色观察褪黑素对CIRI大鼠缺血侧大脑皮质细胞形态学的影响,酶联免疫吸附测定(ELISA)方法测定褪黑素对炎性因子白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的影响。采用Pearson相关性分析结合一般线性回归分析对建模后各时点CIRI组与褪黑素治疗组大鼠缺血侧大脑皮质Iba-1+细胞数差值与胶质纤维酸性蛋白(GFAP+)/AQP4+及GFAP+/NKCC1+细胞数差值进行相关性分析。结果 T2WI和DWI扫描结果显示,Sham组大鼠脑组织信号均匀,未见异常信号。建模后24 h, CIRI组见大范围异常信号脑损伤区,缺血侧大脑半球水肿,大脑�
Objective To investigate the effect of melatonin on cytotoxic edema in the cerebral cortex of cerebral ischemia-reperfusion injury(CIRI)rats and its mechanism.Methods Sixty adult healthy male Sprague Dawley rats without specific pathogens were completely randomized into Sham-operated group(12 rats),CIRI group(24 rats)and melatonin-treated group(24 rats).CIRI group and melatonin-treated group were established CIRI rat models using the modified Zea-Longa′s method.Melatonin(5 mg/kg)was injected intraperitoneally 30 min before and after modeling in the melatonin-treated group,and an equal amount of isotonic saline was injected 30 min before and after modeling in the CIRI group.In the Sham-operated group,the right common carotid artery was isolated and the skin was sutured,and an equal amount of isotonic saline was injected 30 min before and after Sham surgery.At 2,6,12,24 h and 72 h after modeling,in vivo T2-weighted imaging(T2WI)scans and diffusion-weighted imaging(DWI)combined with apparent diffusion coefficient(ADC)maps were performed by MRI to dynamically analyze the changes in percentage of brain infarct volume and percentage of brain edema,and the changes in relative apparent diffusion coefficient(rADC)values in the cerebral cortex of CIRI rats after melatonin intervention.At 6,24 h and 72 h after modeling,the effects of melatonin on apoptosis in the ischemic side of the cerebral cortex of CIRI rats were observed by deoxynucleotide end-transferase-mediated 2′-deoxyuridine 5′-triphosphate-mediated nick end-labeling(TUNEL)method,and the effect of melatonin on water channel protein 4(AQP4)and sodium-potassium-chloride co-transport protein 1(NKCC1)co-labeled with glial fibrillary acidic protein(GFAP)were observed by immunofluorescent double-labeling staining.The effect of melatonin on the activation of microglia labeled with Ionized calcium binding adapter molecule(Iba-1)was investigated by immunofluorescence staining combined with Western Blot(WB).At 24 h after modeling,hematoxylin-eosin(HE)staining was perf
作者
马瑜徽
李宸
李振
张新月
刘晨萌
王灿
肖培伦
王锡臻
王晓莉
Ma Yuhui;Li Chen;Li Zhen;Zhang Xinyue;Liu Chenmeng;Wang Can;Xiao Peilun;Wang Xizhen;Wang Xiaoli(Medical Imaging Center,Affiliated Hospital of Weifang Medical University,Weifang,Shandong 261053,China)
出处
《中国脑血管病杂志》
CAS
CSCD
北大核心
2023年第4期248-260,共13页
Chinese Journal of Cerebrovascular Diseases
基金
国家自然科学基金(82071888)
山东省自然科学基金(ZR2020MH074)
山东省中医药科技发展计划项目(2019-0417)
2019年高等学校青创人才引育计划。
