辣椒素预处理对大鼠脑缺血再灌注损伤的保护作用的代谢组学研究  

Neuroprotective effects of capsaicin pretreatment on ischemic stroke rats revealed by metabolomic approach

作　　者：张懿[1] 朱德生[1] 王艳[1] 

机构地区：[1]上海市脑血管病防治研究所,上海201203

出　　处：《中风与神经疾病杂志》2016年第11期987-994,共8页Journal of Apoplexy and Nervous Diseases

基　　金：上海市自然科学基金(No.14ZR1436700);上海市卫计委科研课题(No.20144Y0225;No.201440347)

摘　　要：目的研究辣椒素预处理对大鼠局部脑缺血再灌注损伤的保护作用。方法采用大脑中动脉栓塞方法建立大鼠脑缺血再灌注损伤模型,选用SD大鼠24只,适应1 w后随机分为3组,假手术组(Sham),模型组(Mod)以及辣椒素预处理组(CAP,0.2 mg/kg),每组8只,采用气相色谱-质谱法(GC-MS)方法分析各组大鼠脑组织皮质样本,运用主成分分析(PCA),偏最小二乘法-判别分析(PLS-DA),正交偏最小二乘法-判别分析(OPLSDA),进行差异化合物筛选和鉴定。结果 OPLS-DA得分图能够较明显的区分各组样本。根据变量重要性投影值(VIP)和载荷图,并结合t检验的P值(P<0.05)来筛选差异表达代谢物。与假手术组大鼠相比,模型组大鼠的皮质代谢谱明显不同,兴奋性氨基酸谷氨酸(P<0.05)、肌醇(P<0.01)等代谢物出现显著增高;花生四烯酸(P<0.01)、异亮氨酸(P<0.001)、甲硫氨酸(P<0.001)、L-半胱氨酸(P<0.001)、天冬氨酸(P<0.05)、苯丙氨酸(P<0.05)等氨基酸、1-磷酸-葡萄糖(P<0.05)、6-磷酸-葡萄糖(P<0.01)等糖代谢物出现显著降低。与模型组大鼠相比,辣椒素预处理组异亮氨酸、甲硫氨酸、L-半胱氨酸、天冬氨酸、苯丙氨酸等氨基酸恢复正常水平,3、5-二羟苯甘氨酸(P<0.001)等代谢物显著增高。结论代谢组学表明,发生脑缺血再灌注损伤时,皮质内能量代谢、神经递质出现紊乱,辣椒素预处理可能通过改善对缺血脑组织的能量代谢障碍,缓解兴奋性谷氨酸的释放量,改善生物膜的功能对脑缺血再灌注损伤发挥保护作用。气相色谱质谱技术能较为全面地反映生物体在药物干预作用下的代谢状态变化,并能通过代谢物含量的差异推断药物的作用机制,为研究脑缺血损伤机制和治疗脑缺血药物提供了新的思路。Objective The purpose of this study is to elucidate the neuroprotective effects of capsaicin pretreatment on ischemic stroke rats revealed by GC-MS metabolomicapproach .Methods The establishment of middle cerebral artery occlusion is a ideal method to mimic cerebral ischemia reperfusion injury .Twenty-four adult SD rats were randomly divided into three groups：sham group （Sham）,model group （Mod） and capsaicin pretreatment group （CAP,0.2 mg/Kg）.The cerebral cortex tissues were collected using GC-MS metabolomic approach and the screening of differential expressed metab-olites were adopted by multivariate statistical analyses including PCA analysis , partial least squares discriminant analysis （ PLS-DA） and orthogonal partial least squares discriminant analysis （ OPLS-DA） .Results OPLS-DA score plot can ob-viously distinguish each group and the screening methods of differentially expressed metabolites were according to the VIP value and the loading scatter plot combined with t-test （P〈0.05）.The analytical results showed that the metabolism spec-trum in model group was different from sham group ,excitatory amino acid glutamic acid （P〈0.05） metabolites were signif-icantly increased while arachidonic acid （P〈0.01）and some amino acid such as isoleucine （P〈0.001）,methionine（P〈0.001）,L-cysteine（P〈0.001）,aspartic acid（P〈0.05）,phenylalanine（P〈0.05） and energy metabolites such as 1-phosphate-glucose（P〈0.05）、6-phosphate-glucose（P〈0.01）significantly decreased.Compared with model group,the metabolism spectrum of capsaicin pretreatment group represents a distinct metabolism spectrum ,inosine,isoleucine,methio-nine,L-cysteine,aspartic acid,phenylalanine and other amino acids were approaching to normal levels ,3,5-dihydroxy phen-yl glycine （P〈0.001） and other metabolites significantly increased .Conclusion These differentially expressed metabo-lites are related to the disturbance in energy metabolism ,glycometabolism .Capsaicin pretreatment ma

关 键 词：TRPV1 辣椒素 脑缺血再灌注损伤 代谢组学 

分 类 号：R743[医药卫生—神经病学与精神病学]