中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (40): 5933-5938.doi: 10.3969/j.issn.2095-4344.2016.40.001

• 脑及脊髓损伤动物模型 Animal models of brain and spinal cord injuries •    下一篇

补阳还五汤对脑出血模型大鼠脑组织PI3K、Akt、Bcl-2、BAX蛋白表达的影响

吴晓光1，仇志富1，孟 杰2，祖炳学3，李蒙蒙4，苗 惠4   

  1. 1河北省中医药抗痴呆重点研究室/承德医学院基础医学研究所，河北省承德市 067000；2承德医学院附属医院，河北省承德市 067000；3抚宁县第二人民医院，河北省秦皇岛市 066300；4承德医学院2012级本科，河北省承德市 067000
  • 修回日期:2016-07-13 出版日期:2016-09-30 发布日期:2016-09-30
  • 作者简介:吴晓光，男，1975年生，满族，硕士，副研究员，主要从事中医药抗痴呆研究。
  • 基金资助:

    河北省卫生厅资助项目资助(2014013)；河北省高等学校重点学科病理学与病理生理学资助项目(2013)；承德医学院重点资助项目(201607)

Effects of Buyanghuanwu decoction on the protein expression of PI3K, Akt, Bcl-2 and BAX in brain tissue of a rat model of cerebral hemorrhage

Wu Xiao-guang1, Qiu Zhi-fu1, Meng Jie2, Zu Bing-xue3, Li Meng-meng4, Miao Hui4   

  1. 1Anti-dementia MedicineKey Laboratory in Hebei Province / Institute of Basic Medicine, Chengde Medical College, Chengde 067000, Hebei Province, China; 2Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China; 3Funing County Second People’s Hospital, Qinhuangdao 066300, Hebei Province, China; 4Grade 2012 Undergraduate Student, Chengde Medical College, Chengde 067000, Hebei Province, China
  • Revised:2016-07-13 Online:2016-09-30 Published:2016-09-30
  • About author:Wu Xiao-guang, Master, Associate researcher, Anti-dementia Medicine Key Laboratory in Hebei Province/Institute of Basic Medicine, Chengde Medical College, Chengde 067000, Hebei Province, China
  • Supported by:

    a grant from the Health Department of Hebei Province, No. 2014013; the Key Pathology and Pathophysiology Subject of Colleges and Universities in Hebei Province, No. 2013; the Key Program of Chengde Medical College, No. 201607

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文题释义：
补阳还五汤：由生黄芪120 g、当归尾6 g、赤芍4.5 g、川芎4.5 g、干地龙3 g、桃仁3 g、红花3 g组成，具有增强微血管抗损伤的能力，减轻脑缺血诱发的脑组织水肿，抑制神经元凋亡的作用。但其能否通过激活PI3K/Ak信号通路，进而调控凋亡相关蛋白的表达，起到抑制细胞凋亡、减轻脑水肿的作用，未见报道。
Rosenberg法建立脑出血大鼠模型：于大鼠头部正中行纵行切口，将大鼠骨膜剥离，使其冠状缝和前囟裸露。在前囟之前约2.0 mm、中线右侧约3.0 mm处钻一约1.0 mm直径的小孔。以微量注射器从股动脉抽取适量的血液，借助立体定向仪沿钻孔垂直进针6.0 mm左右，缓缓注血80 μL，留针10 min后撤针，以消毒针缝合切口。
 
摘要
背景：补阳还五汤具有出色的神经保护作用，能够高效抑制脑缺血再灌注损伤导致的神经细胞凋亡。
目的：研究补阳还五汤对脑出血大鼠血肿周围神经元凋亡的影响及作用机制。
方法：将72只成年SD大鼠随机分为假手术组、模型组、补阳还五汤组、银杏叶片组，除假手术组，其余3组建立大鼠脑出血模型，造模后第2天开始，补阳还五汤组、银杏叶片组分别灌胃给予补阳还五汤26 g/(kg•d)、银杏叶片3.5 mg/(kg•d)，假手术组及模型组灌胃给予等量生理盐水，连续给药14 d。末次给药后，取脑组织，TUNEL法检测神经元凋亡，免疫组织化学法检测PI3K、Akt、Bcl-2、BAX蛋白表达，干湿重法检测脑组织水含量，伊文思蓝法检测血脑屏障通透性。
结果与结论：①与假手术组比较，模型组大鼠脑组织神经元凋亡数目、脑组织水含量、伊文思蓝含量及PI3K、Akt、Bcl-2、BAX蛋白表达增加(P < 0.05)；②与模型组比较，补阳还五汤组、银杏叶片组凋亡神经元数目、BAX蛋白表达、脑组织水含量、伊文思蓝含量显著降低(P < 0.05)，PI3K、Akt、Bcl-2蛋白表达明显升高(P < 0.05)；③结果表明：补阳还五汤可能通过降低血脑屏障通透性，减轻脑水肿，激活PI3K/Akt信号转导通路，进而调控Bcl-2与BAX蛋白表达比值，起到抑制脑出血神经元凋亡、保护脑组织的作用。
 
ORCID: 0000-0002-1057-3158(吴晓光)

关键词: 实验动物, 神经损伤与修复动物模型, 补阳还五汤, 脑出血, PI3K/Akt, Bcl-2, BAX, 组织神经细胞凋亡, 脑水肿, 血脑屏障

Abstract:

BACKGROUND:Buyanghuanwu decoction has excellent neuroprotective effect and can efficiently suppress nerve cell apoptosis caused by cerebral ischemia-reperfusion injury.

OBJECTIVE: To investigate the effect and mechanisms of Buyanghuanwu decoction on neuronal apoptosis around hematoma in cerebral hemorrhage rats.
METHODS: Seventy-two adult Sprague-Dawley rats were randomly divided into sham operation group, model group, Buyanghuanwu decoction group, and Ginkgo biloba group. Except the sham operation group, rat models of cerebral hemorrhage were established in other three groups. At 2 days after modeling, rats in the Buyanghuanwu group and Ginkgo biloba group were given Buyanghuanwu decoction 26 g/(kg • d)and Ginkgo biloba 3.5 mg/(kg • d) daily by gavage, for 14 consecutive days. Rats in the sham operation group and model group received an equal volume of saline for 14 consecutive days. After the last administration, brain tissue was obtained. TUNEL assay was utilized to detect neuronal apoptosis. Immunohistochemistry was used to detect PI3K, Akt, Bcl-2, and BAX protein expression. Wet and dry weight method was used to detect brain water content. Evans Blue assay was utilized to determine blood-brain barrier permeability.
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, the number of apoptotic neurons, brain water content, Evans blue content and PI3K, Akt, Bcl-2, BAX protein expression increased in the model group (P < 0.05). (2) Compared with the model group, the number of apoptotic neurons, BAX protein expression, brain water content and Evans blue content were significantly reduced in the Buyanghuanwu group and Ginkgo biloba group (P < 0.05), but PI3K, Akt and Bcl-2 protein expression was significantly increased (P < 0.05). (3) Results suggested that Buyanghuanwu decoction inhibited neuronal apoptosis and protected brain tissue by reducing blood-brain barrier permeability, cerebral edema, and by activating PI3K/Akt signaling pathway, regulating Bcl-2 and BAX protein expression ratio.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Cerebral Hemorrhage, Drugs, Chinese Herbal, Models, Animal, Tissue Engineering

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