摘要
目的:探究电针对新生缺氧缺血性脑损伤(HIBD)大鼠的保护作用及机制。方法:将新生SD大鼠随机分为假手术(Sham)组、HIBD组、电针组、电针+TXNIP NC组和电针+TXNIP OE组,除Sham组外,其余各组采用改良RICE法建立大鼠HIBD模型;电针组、电针+TXNIP NC组和电针+TXNIP OE组大鼠进行电针治疗,电针+TXNIP NC组、电针+TXNIP OE组大鼠分别经脑室注射TXNIP NC CRISPR质粒、TXNIP OE CRISPR质粒溶液,电针组注射无菌生理盐水;治疗周期完成后,采用莫里斯水迷宫测试大鼠学习记忆能力;红四氮唑(TTC)法检测脑梗死体积,尼氏(Nissl)和苏木素-伊红(HE)染色观察海马神经细胞形态;免疫荧光法检测海马区细胞焦亡;蛋白免疫印迹法检测大鼠海马组织含半胱氨酸的天冬氨酸蛋白水解酶-1(cleaved-caspase-1)、天冬氨酸蛋白水解酶-1前体(pro-caspase-1)、凋亡相关的斑点样蛋白(ASC)、硫氧还蛋白互作蛋白(TXNIP)、Nod样受体蛋白3(NLRP3)表达;酶联免疫吸附法检测大鼠海马组织白细胞介素(IL)-18、IL-1β水平。结果:Sham组无脑梗死,海马区神经细胞排列规则致密;与Sham组比较,HIBD组海马区神经细胞排列错乱疏松,脑梗死体积、逃避潜伏期、海马区cleaved-caspase-1+TUNEL+细胞、海马组织cleaved-caspase-1/pro-caspase-1、ASC、TXNIP、NLRP3蛋白及IL-18、IL-1β水平明显增加,穿越平台的次数明显减少(P<0.05);与HIBD组比较,电针组、电针+TXNIP NC组海马区细胞损伤明显改善,脑梗死体积、逃避潜伏期、海马区cleaved-caspase-1+TUNEL+细胞、海马组织cleaved-caspase-1/pro-caspase-1、ASC、TXNIP、NLRP3蛋白及IL-18、IL-1β水平减少,穿越平台的次数增加(P<0.05);与电针组、电针+TXNIP NC组比较,电针+TXNIP OE组海马区细胞损伤加重,脑梗死体积、逃避潜伏期、海马区cleaved-caspase-1+TUNEL+细胞、海马组织cleaved-caspase-1/pro-caspase-1、ASC、TXNIP、NLRP3蛋白及IL-18、IL-1β水平增加,穿越平台的次数减少(
Objective:To explore the protective effect and mechanism of electroacupuncture on neonatal hypoxic ischemic brain damage(HIBD)rats.Methods:Neonatal SD rats were randomly divided into sham operation group,HIBD group,electroacupuncture group,electroacupuncture+TXNIP NC group and electroacupuncture+TXNIP OE group,HIBD model was established by modified RICE method in all but the sham groups.Rats in electroacupuncture group,electroacupuncture+TXNIP NC group and electroacupuncture+TXNIP OE group were treated with electroacupuncture.Rats in electroacupuncture+TXNIP NC group and electroacupuncture+TXNIP OE group were injected with TXNIP NC CRISPR plasmid and TXNIP OE CRISPR plasmid solution via cerebral ventricle respectively.Rats in electroacupuncture group were injected with sterile normal saline.At the end of the treatment cycle,Morris water maze was used to test the abilities of learning and memory;triphenyltetrazolium chloride(TTC)was used to measure the volume of cerebral infarction,Nissl and hematoxylin-eosin(HE)staining were used to observe the morphology of hippocampal neurons;immunofluorescence was used to detect the pyroptosis of hippocampal cells;western blot was used to detected the expressions of cleaved cysteinyl aspartate specific proteinase-1(cleaved caspase-1),pro-cysteinyl aspartate specific proteinase-1(pro-caspase-1),apotosis-associated speck-like protein(ASC),thioredoxin interacting protein(TXNIP)and Nod-like receptor protein 3(NLRP3)in rat hippocampus;enzyme linked immunosorbent assay was used to detect the expression of interleukin(IL)-18 and IL-1βin rat hippocampus.Results:There was no cerebral infarction in the control group,and the arrangement of neurons in hippocampus was regular and dense.Compared with sham group,HIBD group had disordered and loose arrangement of nerve cells in hippocampus,cerebral infarction volume,escape latency,cleaved-caspase-1+TUNEL+cells in hippocampus,cleaved-caspase-1/pro-caspase-1,ASC,TXNIP,NLRP3 protein,IL-18,IL-1βlevels were significantly increased,and times of c
作者
邓寒冰
李春琴
张粲
金祖敏
DENG Hanbing(Haikou Third People's Hospital, Hainan Haikou 571100, China)
出处
《河北医学》
CAS
2021年第8期1233-1239,共7页
Hebei Medicine
基金
云南省应用基础研究联合专项项目,(编号:201701YH0037)。
