丁苯酞通过抑制神经元自噬保护脑缺血再灌注损伤的机制研究  

Mechanism study on the protective effect of butylphthalein on cerebral ischemia-reperfusion injury by inhibiting neuronal autophagy

作　　者：徐翔 陆晓培 黄玉钿 吴佳琳 Xu Xiang;Lu Xiao-pei;Huang Yu-dian;Wu Jia-lin(Department of Neurology,Fuzhou General Hospital of Nanjing Military Region,Fuzhou 350009,China)

机构地区：[1]福州市第一总医院神经内科,福建福州350009

出　　处：《北方药学》2025年第1期10-15,共6页Journal of North Pharmacy

基　　金：福建省自然科学基金项目(编号:2022J011306)。

摘　　要：目的:探究丁苯酞通过抑制神经元自噬保护脑缺血再灌注损伤(Cerebral Ischemia-Reperfusion Injury,IRI)的相关机制。方法:选取健康雄性SD大鼠80只,随机分为5组(n=16),即对照组(Con组)、改良Zea-longa线栓法制备大鼠右侧大脑中动脉局灶性脑缺血再灌注模型即MCA组、建模前腹腔内注射10 mg/kg消旋3-正丁基苯酞/NBP为MCA+NBP组、建模前腹腔注射5 mg/kg N-乙酰半胱氨酸/NAC即MCA+NAC组、建模前腹腔注射NBP+NCA即MCA+NBP+NCA组,其中MCA组、MCA+NAC组、MCA+NBP+NCA组再灌注24 h。采用改良Zea-longa评分法评估各组大鼠神经功能;制作大鼠脑组织切片,氯化三苯基四氮唑(Triphenyltetrazolium Chloride,TTC)染色计算脑梗死体积;经电子显微镜及免疫荧光染色评估大鼠大脑皮层神经元形态;蛋白质印迹法测定大鼠脑组织中P62的蛋白表达量。结果:各组大鼠存活率依次为Con组(100%)、MCA组(56.25%)、MCA+NCA组(62.50%)、MCA+NBP+NCA组(93.75%)。相较于MCA组,MCA+NBP与MCA+NBP+NCA组神经功能评分均显著下降(P<0.05),MCA与MCA+NCA组神经功能评分差异无统计学意义(P>0.05)。相较于Con组,其余4组大鼠脑组织均出现梗死灶;与MCA组相比,MCA+NBP及MCA+NBP+NCA组脑梗死体积均显著下降(P<0.05)。与Con组相比,光镜下观察脑组织神经元自噬小体,MCA组神经元细胞中重度水肿、细胞核固缩、线粒体呈球状肿胀、自噬溶酶体数量增加;而MCA+NBP及MCA+NBP+NCA组神经元细胞轻中度水肿,结构完整。与Con组相比,MCA组P62的表达显著升高(P<0.05);与MCA组相比,MCA+NBP及MCA+NBP+NCA组P62的表达均下降(P<0.05),而MCA与MCA+NCA组P62表达量无统计学差异(P>0.05)。结论:丁苯酞(NBP)可抑制神经元过度自噬、降低P62表达。从存活率看,NBP+NCA联合使用效果更佳。在神经功能评分、脑梗死体积、神经元形态及P62表达方面,NBP单独或与NCA联合使用均有积极效果,对脑缺血再灌注损伤有保护作用。Objective:To investigate the mechanism by which butylphthalein protects against cerebral ischemia-reperfusion injury(IRI)by inhibiting neuronal autophagy.Method:Eighty healthy male SD rats were selected and randomly divided into five groups(n=16):control group(Con group),modified Zea longa suture method to prepare a rat model of focal cerebral ischemia-reperfusion in the right middle cerebral artery(MCA group),pre modeling intraperitoneal injection of 10 mg/kg raceme-3-n-butylphthalide/NBP as MCA+NBP group,pre modeling intraperitoneal injection of 5 mg/kg N-acetylcysteine/NAC as MCA+NAC group,pre modeling intraperitoneal injection of NBP+NCA as MCA+NBP+NCA group.Among them,MCA group,MCA+NAC group,and MCA+NBP+NCA group were reperfusion for 24 hours.Evaluate the neurological function of rats in each group using the improved Zea longa scoring method;Prepare rat brain tissue slices and calculate the volume of cerebral infarction by staining with Triphenyltetrazolium Chloride(TTC);Evaluate the morphology of rat cortical neurons using electron microscopy and immunofluorescence staining;The protein expression level of P62 in rat brain tissue was determined by Western blotting.Results:The survival rates of each group of rats were as follows:Con group(100%),MCA group(56.25%),MCA+NCA group(62.50%),and MCA+NBP+NCA group(93.75%).Compared with the MCA group,both the MCA+NBP and MCA+NBP+NCA groups showed a significant decrease in neurological function scores(P<0.05),while there was no statistically significant difference in neurological function scores between the MCA and MCA+NCA groups(P>0.05).Compared to the Con group,the brain tissues of the other four groups of rats showed infarcted lesions;Compared with the MCA group,both the MCA+NBP and MCA+NBP+NCA groups showed a significant decrease in cerebral infarction volume(P<0.05).Compared with the Con group,under light microscopy,autophagosomes in brain tissue neurons were observed.In the MCA group,there was severe edema,nuclear condensation,spherical swelling of mitochondria,an

关 键 词：脑缺血再灌注损伤 丁苯酞 神经元保护 自噬 

分 类 号：R-332[医药卫生]