井穴放血对急性高原缺氧脑损伤大鼠海马线粒体自噬相关蛋白表达的影响  

Effect of blood-letting puncture of “Well-points” on hippocampal mitophagy-related protein expression in rats with acute brain injury due to hypobaric hypoxia

作　　者：黄雅倩 李梦馨 王超 李永平[1] HUANG Ya-qian;LI Meng-xin;WANG Chao;LI Yong-ping(Department of Chinese Medicine,Medical College of Qinghai University,Xining 810001,China)

机构地区：[1]青海大学医学院中医系,西宁810001

出　　处：《针刺研究》2021年第4期301-305,共5页Acupuncture Research

基　　金：青海省科技厅应用基础研究项目(No.2020-ZJ-760);全国中医药创新骨干人才培训项目(国中医药人教函[2019]128号);青海省中藏医药(循证)研究课题项目(No.2019106)。

摘　　要：目的:观察井穴放血对急性高原缺氧脑损伤大鼠海马线粒体自噬相关蛋白表达的影响,探讨井穴放血治疗急性高原低氧脑损伤的生物学机制。方法:雄性SD大鼠随机分为正常对照组、低压低氧组、井穴放血干预组,后两组又按6、12、24、48、72 h分为5个亚组,每组9只。采用实验动物低压模拟舱(海拔5 000 m)制备急性高原低氧脑损伤模型。井穴放血干预组大鼠按"少商""商阳""中冲""关冲""少冲""少泽"的顺序进行放血预处理,每日1次,共7 d。采用HE染色法观察大鼠海马组织变化;ELISA法检测血清缺氧诱导因子(HIF)-1α、血管内皮生长因子(VEGF)含量;Western blot法检测大鼠海马组织Beclin-1、LC3Ⅱ蛋白的表达情况。结果:与正常对照组比较,低压低氧组各时点海马组织有不同程度损伤,各时点血清HIF-1α、VEGF含量升高(P<0.01),12、24 h时海马组织Beclin-1蛋白表达升高(P<0.05),各时点海马组织LC3Ⅱ蛋白表达均升高(P<0.05,P<0.01)。与同时点低压低氧组比较,井穴放血干预组大鼠海马组织损伤程度较轻,各时点血清HIF-1α、VEGF含量升高(P<0.01,P<0.05),12 h时海马组织Beclin-1蛋白表达升高(P<0.05), 24、48、72 h时海马组织LC3Ⅱ蛋白表达明显升高(P<0.01)。结论:井穴放血干预可改善急性高原缺氧大鼠海马组织损伤,其机制可能与上调血清HIF-1α、VEGF含量及海马组织中线粒体自噬相关蛋白Beclin-1、LC3表达有关。Objective To study the effect of blood-letting puncture at "Well-points" of the twelve meridians on hippocampal mitophagy of hypobaric hypoxia-induced brain injury(HHIBI) rats, so as to explore its biological mechanisms underlying improvement of high altitude hypoxia-induced brain injury. Methods Male SD rats were randomly divided into normal control group(n=9), and model and blood-letting groups which were further divided into 6, 12, 24, 48 and 72 h subgroups(n=9 in each subgroup). The HHIBI model was established by putting the rats into a hypobaric hypoxia chamber(equivalent to 5 000 m above sea level).The blood-letting groups were given blood-letting therapy at "Shaoshang"(LU11), "Shangyang"(LI1), "Zhongchong"(PC9), "Guanchong"(SJ1), "Shaochong"(HT9), "Shaoze"(SI9), once a day for 7 days. H.E. staining was used to observe the histopatholo-gical changes of hippocampus tissue. Serum hypoxia inducible factor(HIF)-1α and vascular endothelial growth factor(VEGF) contents were assayed using ELISA, and the expression levels of hippocampal Beclin-1 and LC3-Ⅱ proteins detected using Western blot. Results Compared with the normal control group, the levels of serum HIF-1α and VEGF at each time point, and the expressions of hippocampal Beclin-1 at 12 and 24 h, LC3-Ⅱat each time point were significantly increased in the model group(P<0.05, P<0.01);while in comparison with the model group, the levels of serum HIF-1α and VEGF contents, and the expressions of Beclin-1 at 12 h, LC3-Ⅱ at 24, 48 and 72 h were further significantly up-regulated in the blood-letting group(P<0.01, P<0.05). H.E. staining revealed that the pyramidal cells in the hippocampal CA1 region had a disordered arrangement, and some of them presented swelling with loose and pale cytoplasm or vacuolation at 6, 12 and 24 h, and showed indistinct nucleolus, irregular shape, pyknosis and deep staining and an obvious edema at 48 and 72 h, which was relatively milder in the blood-letting group.Conclusion Blood-letting of "Well-points" can up-regulate serum

关 键 词：放血预处理 十二井穴 高原缺氧 脑损伤 线粒体自噬 

分 类 号：R245.312[医药卫生—针灸推拿学]