适度酒精预适应对大鼠局灶性脑缺血再灌注损伤保护作用的研究

摘要/Abstract

摘要：

目的研究适度酒精预适应对大鼠局灶性脑缺血再灌注诱导的神经元损伤的保护作用。方法将36只雄性SD大鼠随机分为假手术组、缺血再灌注组和酒精预适应组，每组12只。局灶性脑缺血再灌注模型采用右侧大脑中动脉闭塞方式，缺血2 h，再灌注24 h。酒精预适应组在脑缺血再灌注前24 h用95%酒精与0.3 ml无菌蒸馏水混合后进行灌胃，95%酒精体积（μl）计算为：[大鼠体重（g） x0.6]+0.3。其余两组用同等剂量的生理盐水灌胃。每组取8只大鼠在缺血再灌注后24 h进行神经功能评分和氯化2，3，5-三苯基四氮唑（2，3，5-Triphenyltetrazolium chloride，TTC）染色并根据染色结果计算脑梗死体积。每组剩余的4只，在缺血再灌注后24 h进行磁共振T2加权像（T2-weighted imaging，T2WI）序列扫描，然后将大鼠处死取脑，经过冷冻切片后，每只大鼠取第一躯体感觉皮质区的2张切片，1 张用末端转移酶介导的脱氧尿嘧啶核苷三磷酸缺口末端标记（terminal deoxynucleotidyl transferasemediated 2'-deoxyuridine 5'-triphosphate nick-end labeling，TUNEL）检测其凋亡性细胞死亡情况，另一张用Fluoro-Jade B检测其神经元退化变性情况。结果相较于缺血再灌注组，经过适度酒精预适应后会显著降低大鼠的神经功能评分[15.00 （14.25，16.00）vs 3.50（2.25，4.00）（P <0.001）]、脑梗死体积[TTC：（242.80±17.44）mm3 vs （54.83±13.43）mm3；T2WI ：（296.80±8.53）mm3 vs（59.68±9.97）mm3，P均<0.001]、凋亡细胞所占百分比[（33.47±2.23）% vs（9.66±0.84）%，P <0.001]和退化变性神经元所占百分比（[ 45.31±3.40）% vs（23.26±1.25）%，P <0.001]。结论适度酒精预适应可以保护局灶性脑缺血再灌注诱导的神经元损伤。

文章导读： 本研究发现适度酒精预适应对局灶性脑缺血再灌注损伤存在保护作用，为红酒的适量饮用可产生脑预适应保护，预防缺血性卒中提供理论依据。

关键词: 酒精预适应; 脑缺血/再灌注; 神经保护

Abstract:

Objective To study the protective effects of moderate ethanol preconditioning on neuronal injury induced by focal cerebral ischemic-reperfusion in rats. Methods A total of 36 male Sprague-Dawley rats were randomly divided into 3 groups (n =12): (1) sham control group; (2) ischemic-reperfusion group (I/R); (3) ethanol preconditioning group (EtOH-PC). Rats were subjected to 2 h of right middle cerebral artery occlusion (MCAO) and 24 h of reperfusion. EtOH-PC was induced by gavaging animals with a moderate dose of ethanol [volume of ethanol in microliters calculated from the equation (body weight (in g) × 0.6) + 0.3] 24 h before ischemia. This volume of ethanol (95% in μl) was mixed in 0.3 ml of sterile distilled water before gavage. Animals in the sham control group (no I/R) and the I/R-alone group (no EtOH-PC) received a similar volume of saline by gavage. Eight rats of each group were used to perform neurological score and 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining 24 h after ischemia and reperfusion. The volume of cerebral infarction was calculated based on the staining result. Another 4 rats of each group were used to perform T2-weighted imaging (T2WI) MRI 24 h after ischemia and reperfusion. Then rats were killed. After frozen section, two slices was acquired from each rat’s primary somatosensory cortex areas. One slice was used to detect the apoptotic cell by means of terminal deoxynucleotidyl transferase-mediated 2’-deoxyuridine 5’-triphosphate nick-end labeling (TUNEL); the other was used to detect the degeneration of neurons through Fluoro-Jade B. Results Compared to ischemic-reperfusion group, the neurological score (15.00 [14.25, 16.00] vs 3.50 [2.25, 4.00]) (P <0.001), cerebral infarction volume (TTC: [242.80±17.44]mm3 vs [54.83±13.43]mm3; T2WI:[296.80±8.53]mm3 vs [59.68±9.97]mm3) (P <0.001), the percentage of apoptotic cell death ([33.47±2.23]% vs [9.66±0.84]%) (P <0.001) and the percentage of degenerative neurons ([45.31±3.40]% vs [23.26±1.25]%) (P <0.001) were significantly decreased after ethanol preconditioning. Conclusion Moderate ethanol preconditioning can protect against the neuron injury induced by focal cerebral ischemic-reperfusion.

Key words: Ethanol preconditioning; Cerebral ischemia/reperfusion; Neuroprotection

赵一龙，郭安臣，苏芳，李巍巍，杨华俊，孙明，王拥军，王群. 适度酒精预适应对大鼠局灶性脑缺血再灌注损伤保护作用的研究[J]. 中国卒中杂志, 2016, 11(05): 378-385.

ZHAO Yi-Long, GUO An-Chen, SU Fang, et al.. Study on Protective Effects of Moderate Ethanol Preconditioning Against Brain Injury-induced by Focal Cerebral Ischemia / Reperfusion in Rats[J]. Chinese Journal of Stroke, 2016, 11(05): 378-385.

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