电针对缺氧缺血性脑损伤大鼠海马神经元自噬及IGF-1/PI3K/Akt通路的影响  

Effects of electroacupuncture on hippocampal neuron autophagy and IGF-1/PI3K/Akt pathway in rats with hypoxic-ischemic brain damage

作　　者：曹娟 王红怡 王丽欣 陈小宁 林秋 杨庆红 陈东追 符玉秀 CAO Juan;WANG Hongyi;WANG Lixin;CHEN Xiaoning;LIN Qiu;YANG Qinghong;CHEN Dongzhui;FU Yuxiu(Hainan Modern Women and Children's Hospital,Haikou,Hainan,571100;Teaching and Research Section of Immunology of College of Basic Medicine and Life Sciences of Hainan Medical University,Haikou,Hainan,571100)

机构地区：[1]海南现代妇女儿童医院,海南海口571100 [2]海南医学院基础医学与生命科学学院免疫学教研室,海南海口571100

出　　处：《实用临床医药杂志》2022年第14期38-45,共8页Journal of Clinical Medicine in Practice

基　　金：海南省卫生健康行业科研项目(20A200336)。

摘　　要：目的探讨电针对缺氧缺血性脑损伤(HIBD)新生大鼠海马神经元自噬及胰岛素样生长因子1(IGF-1)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法将108只新生大鼠随机分为假手术组、模型组、IGF-1组(0.2 mg/kg)、电针组、电针联合LY294002组(电针+PI3K抑制剂0.3 mg/kg),每组18只。采用左颈总动脉结扎和低氧处理2 h的方法构建新生大鼠HIBD模型(造模成功率为80%)。各组大鼠在手术清醒后和电针治疗结束后进行神经功能缺陷评分;酶联免疫吸附测定(ELISA)检测脑组织IGF-1的含量;苏木精-伊红(HE)染色观察脑组织病理学变化;透射电镜观察细胞自噬情况;免疫荧光双标法检测自噬标志物与神经元特异性核蛋白(NeuN)的共定位表达;Western blot法检测海马组织PI3K/Akt通路、自噬相关蛋白的表达[PI3K、磷酸化PI3K(p-PI3K)、Akt、磷酸化p-Akt(p-Akt)、Beclin-1、微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)、微管相关蛋白轻链3Ⅰ(LC3-Ⅰ)]。结果与假手术组相比,HIBD模型组大鼠海马中神经元损伤加重,神经功能缺损评分、海马组织Beclin-1和LC3-Ⅱ/LC3-Ⅰ表达增加,脑组织IGF-1的含量以及海马组织p-PI3K/PI3K、p-Akt/Akt的表达降低;与模型组相比,IGF-1组、电针组神经元损伤减轻,神经功能缺损评分、Beclin-1和LC3-Ⅱ/LC3-Ⅰ、自噬小体数量、LC3/NeuN共表达的神经元减少或降低,脑组织IGF-1的含量、p-PI3K/PI3K、p-Akt/Akt的表达升高;上述差异均有统计学意义(P<0.05)。LY294002能显著降低IGF-1的水平,减弱电针对海马组织IGF-1/PI3K/Akt通路的激活和海马神经元自噬的抑制作用。结论电针可能通过激活IGF-1/PI3K/Akt通路,抑制海马神经元过度自噬,减轻新生大鼠HIBD。Objective To explore the effects of electroacupuncture on hippocampal neuron autophagy and insulin-like growth factor 1(IGF-1)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)pathway in neonatal rats with hypoxic-ischemic brain damage(HIBD).Methods A total of 108 neonatal rats were randomly divided into sham operation group,model group,IGF-1 group(0.2 mg/kg),electroacupuncture group,electroacupuncture combined with LY294002 group(electroacupuncture combined with 0.3 mg/kg PI3K inhibitor),with 18 rats in each group.HIBD model of neonatal rats was established by ligation of left common carotid artery and hypoxia treatment for 2 hours(the success rate of modeling was 80%).The rats in each group were scored for neurological deficits at being awake after the operation and after electroacupuncture treatment;ELISA was used to detect the content of IGF-1 in brain tissues;hematoxylin-eosin(HE)staining was used to observe the pathological change of brain tissues;transmission electron microscope was used to observe the cell autophagy;immunofluorescence double-labeling method was used to detect the colocalization expression of autophagy markers and neuron-specific nuclear protein(NeuN);western blot method was used to detect the expression of PI3K/Akt pathway and autophagy-related proteins in hippocampus[PI3K,phosphorylated PI3K(p-PI3K),Akt,phosphorylated Akt(p-Akt),Beclin-1,microtubule-related protein light chain 3Ⅱ(LC3-Ⅱ),and microtubule-related protein light chain 3Ⅰ(LC3-Ⅰ)].Results Compared with the sham operation group,the neuron damage in the hippocampus of the HIBD model group was aggravated,the neurological deficit score,the Beclin-1 and LC3-Ⅱ/LC3-I expression in the hippocampus were increased,while the IGF-1 content in brain tissues,p-PI3K/PI3K and p-Akt/Akt expression in hippocampus were reduced;compared with the model group,the neuron damage in the IGF-1 group and the electroacupuncture group was alleviated,the neurological deficit score,Beclin-1 and LC3-Ⅱ/LC3-Ⅰ,the number of autophagosomes a

关 键 词：电针 缺血缺氧脑损伤 自噬 胰岛素样生长因子1 磷脂酰肌醇3激酶 蛋白激酶B 

分 类 号：R743.31[医药卫生—神经病学与精神病学] R245[医药卫生—临床医学]