摘要
目的探讨井穴放血通过调控PI3K/AKT/mTOR信号通路介导的线粒体自噬对急性高原低氧脑损伤的保护作用及分子机制,为临床使用井穴放血防治急性高原低氧脑损伤提供有效靶点及理论依据。方法雄性SD大鼠随机分为对照组(Control组)和实验组,实验组分为模型组(Model组)、井穴放血组(Bloodletting acupuncture at jing-well points组,BAJP组)、非穴位放血组(Bloodletting acupuncture at nonacupoint组,BANA组),每组15只。低压氧舱减压至6000 m海拔高度,将各实验组大鼠低压低氧处理72 h复制急性高原低氧脑损伤大鼠模型。入舱前7天,BAJP组大鼠按照少商、商阳、中冲、关冲、少冲、少泽的顺序点刺放血,每天1次;BANA组大鼠每日剪尾尖放血,两组放血量均为15-20μL。HE染色观察脑组织形态结构变化;透射电镜观察大鼠神经元细胞内线粒体损伤及自噬体形成情况;TUNEL观察海马组织细胞凋亡情况;Western blot检测PI3K、AKT、mTOR表达水平;PCR检测AKT、mTOR mRNA表达水平。结果①与Control组相比,急性高原低氧(Acute high-altitude hypoxia,AHH)大鼠海马组织CA1区变性坏死椎体细胞数量明显增多、线粒体肿胀、自噬体出现、海马组织细胞凋亡数量增加;经过井穴放血治疗后,AHH大鼠各种脑损伤表现均得到缓解;非穴位放血对AHH大鼠脑损伤无明显改善作用。②Western blot检测AHH大鼠海马组织内PI3K、AKT、mTOR磷酸化水平相较于Control组大鼠明显下降(P<0.01),经过井穴放血治疗后,3个分子磷酸化水平进一步下降(P<0.01),非穴位放血治疗未对这些分子的磷酸化水平产生显著影响(P>0.05),AKT、mTOR mRNA表达水平进一步证明了上述趋势。结论井穴放血可以对急性高原低氧脑损伤起到保护作用,具有穴位特殊性,其机制可能与抑制PI3K/AKT/mTOR通路促进线粒体自噬水平升高,改善线粒体生理,增强机体抗细胞凋亡、耐缺氧能力有关。
Objective To investigate the protective effect and molecular mechanism of bloodletting acupuncture at Jing-well points on acute high-altitude hypoxia brain injury through regulating PI3K/AKT/mTOR signaling pathway-mediated mitochondrial autophagy,and to provide an effective target and theoretical basis for the clinical use of bloodletting acupuncture at Jing-well points to prevent and treat acute high-altitude hypoxia brain injury.Methods Male SD rats were randomly divided into Control group(n=15)and experimental group,and the experimental group was divided into Model group(n=15),Bloodletting Acupuncture at Jing-well Points of hand group(BAJP group,n=15),Bloodletting Acupuncture at Non-Acupoint group(BANA group,n=15).The low pressure oxygen chamber was depressurized to 6000 m altitude,and the rats in each experimental group were treated with low-pressure hypoxia for 72 h to replicate the acute high-altitude hypoxia brain injury rat model.The rats in the BAJP group were bled according to the order of"Shaoshang"(LU11),"Shangyang"(LI1),"Zhongchong"(PC9),"Guanchong"(SJ1),"Shaochong"(HT9),"Shaoze"(SI1),once a day for 7 days.The rats in the BANA group were bled by cutting the tail tip daily,and the amount of blood bled was 15-20μL in both groups.The expression levels of PI3K,AKT and mTOR in hippocampal tissues of rats were detected by Western blot;AKT and mTOR mRNA expression levels in hippocampal tissues of rats were detected by PCR.Results Compared with the Control group,the number of degenerative necrotic vertebral cells in CA1 area of hippocampal tissue,swelling of mitochondria,appearance of autophagosomes,and increase of apoptosis in hippocampal tissue of Acute High-altitude Hypoxia(AHH)rats were significantly increased;After bloodletting acupuncture at Jing-well points of hand treatment,various brain injury manifestations in AHH rats were alleviated;Bloodletting acupuncture at non-acupoint had no significant ameliorating effect on AHH rats'brain injury.Western blot detected a significant decrease in the phosphor
作者
李梦馨
王超
童丽
洒玉萍
任延明
李永平
Li Mengxin;Wang Chao;Tong Li;Sa Yuping;Ren Yanming;Li Yongping(Department of Chinese Medicine,Medical College of Qinghai University,Qinghai Provincial Key Laboratory of Traditional Chinese Medicine Research for Glucolipid Metabolic Diseases,Xining 810001,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第7期2509-2517,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
青海省科学技术厅应用基础研究项目(2020-ZJ-760):基于mTOR-PI3K-Atg信号通路介导的线粒体自噬研究井穴放血对急性高原缺氧模型大鼠脑损伤的保护作用及其分子机制
负责人:李永平。
关键词
高原低氧
脑损伤
井穴放血
线粒体自噬
PI3K/AKT/MTOR
Acute high-altitude hypoxia
Brain injury
Bloodletting acupuncture at Jing-well points
Mitochondrial autophagy
PI3K/AKT/mTOR
