醒脑静注射液对蛛网膜下腔出血大鼠学习记忆能力及海马区自噬相关蛋白表达的影响
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摘要
目的探讨醒脑静注射液对蛛网膜下腔出血(SAH)大鼠学习记忆能力及海马区自噬相关蛋白表达的影响。方法将36只大鼠随机分为假手术组、模型组、醒脑静组各12只。模型组、醒脑静组采用颈动脉穿刺法制作SAH大鼠模型,假手术组仅将纤芯沿血管穿入颅内但不刺破血管。造模成功后,醒脑静组给予醒脑静注射液2 m L腹腔注射,1次/12 h;假手术组和模型组给予等量生理盐水腹腔注射。术后24 h进行穿梭箱实验,比较三组大鼠的逃避反应时间和逃避反应次数;处死大鼠,取海马组织,HE染色观察神经细胞形态,免疫组化法检测Beclin-1、LC3Ⅱ阳性神经细胞个数,Western blotting法检测海马组织中自噬特异性标志物Beclin-1、LC3Ⅱ蛋白表达。结果与假手术组比较,模型组穿梭箱实验逃避反应时间延长,逃避反应次数减少(P均<0. 05);与模型组比较,醒脑静组逃避反应时间缩短,逃避反应次数增加(P均<0. 05)。假手术组海马神经细胞多数形态正常;模型组胞核梭形固缩、深染情况明显增多,并有核破碎现象;醒脑静组与模型组比较,胞核梭形固缩、深染、核破碎情况明显减少。模型组海马Beclin-1、LC3Ⅱ阳性神经细胞数较假手术组增多,醒脑静组较模型组增多(P均<0. 05)。与假手术组比较,模型组海马组织中Beclin-1、LC3Ⅱ蛋白表达升高(P均<0. 05);与模型组比较,醒脑静组Beclin-1、LC3Ⅱ蛋白表达升高(P均<0. 05)。结论醒脑静可改善SAH大鼠的学习记忆能力,保护脑神经功能,其机制可能与激活SAH大鼠海马区神经细胞自噬相关蛋白表达有关。
Objective To investigate the effect of Xingnaojing injection on learning and memory ability and autophagy-related protein expression of hippocampus nerve cells in rats with subarachnoid hemorrhage( SAH). Methods Thirtysix rats were randomly divided into the sham operation group,model group and Xingnaojing group,with 12 rats in each group. The SAH rat models were made by carotid puncture; in the sham operation group,we only penetrated the core into the skull along the blood vessel,but did not pierce the blood vessel. After successful modeling,rats in the Xingnaojing group were given intracerebroventricular injection of Xingnaojing 2 m L,1 time/12 h; rats in the sham operation group and model group were given the same amount of normal saline. The shuttle box training was performed at 24 h after surgery,and we compared the time of avoidance reaction and the times of avoidance reaction in three groups. Then the rats were sacrificed and hippocampus were taken,we observed the nerve cell morphology by HE staining,using immunohistochemistry to detect the number of Beclin-1 and LC3Ⅱ positive cells,and Western blotting to detect the expression of autophagy-specific markers Beclin-1 and LC3Ⅱ protein. Results Compared with the sham operation group,the time of avoidance reaction was prolonged,and the times of avoidance decreased in the model group( both P < 0. 05). Compared with the model group,the time of avoidance reaction was shortened,and the times of avoidance increased in the model group( both P <0. 05). Compared with the sham operation group,the number of hippocampal nerve cells with normal morphology decreased in the model group( P < 0. 05). Compared with the model group,the number of hippocampal nerve cells with normal morphology increased in the Xingnaojing group( P < 0. 05). Compared with the sham operation group,the number of Beclin-1 and LC3Ⅱ positive cells increased in the model group,and the Xingnaojing group was higher than the model group( all P< 0. 05). Compared with the sham operation group,the expression of Beclin-1 and LC3Ⅱ protein increased in the model group( both P < 0. 05). Compared with the model group,the expression of Beclin-1 and LC3Ⅱ protein increased in Xingnaojing group( both P < 0. 05). Conclusion Xingnaojing can improve the learning and memory ability of SAH rats and protect brain nerve function,and its mechanism may be related to the activation of autophagy-related protein expression in hippocampus of SAH rats.
Objective To investigate the effect of Xingnaojing injection on learning and memory ability and autophagy-related protein expression of hippocampus nerve cells in rats with subarachnoid hemorrhage( SAH). Methods Thirtysix rats were randomly divided into the sham operation group,model group and Xingnaojing group,with 12 rats in each group. The SAH rat models were made by carotid puncture; in the sham operation group,we only penetrated the core into the skull along the blood vessel,but did not pierce the blood vessel. After successful modeling,rats in the Xingnaojing group were given intracerebroventricular injection of Xingnaojing 2 m L,1 time/12 h; rats in the sham operation group and model group were given the same amount of normal saline. The shuttle box training was performed at 24 h after surgery,and we compared the time of avoidance reaction and the times of avoidance reaction in three groups. Then the rats were sacrificed and hippocampus were taken,we observed the nerve cell morphology by HE staining,using immunohistochemistry to detect the number of Beclin-1 and LC3Ⅱ positive cells,and Western blotting to detect the expression of autophagy-specific markers Beclin-1 and LC3Ⅱ protein. Results Compared with the sham operation group,the time of avoidance reaction was prolonged,and the times of avoidance decreased in the model group( both P < 0. 05). Compared with the model group,the time of avoidance reaction was shortened,and the times of avoidance increased in the model group( both P <0. 05). Compared with the sham operation group,the number of hippocampal nerve cells with normal morphology decreased in the model group( P < 0. 05). Compared with the model group,the number of hippocampal nerve cells with normal morphology increased in the Xingnaojing group( P < 0. 05). Compared with the sham operation group,the number of Beclin-1 and LC3Ⅱ positive cells increased in the model group,and the Xingnaojing group was higher than the model group( all P< 0. 05). Compared with the sham operation group,the expression of Beclin-1 and LC3Ⅱ protein increased in the model group( both P < 0. 05). Compared with the model group,the expression of Beclin-1 and LC3Ⅱ protein increased in Xingnaojing group( both P < 0. 05). Conclusion Xingnaojing can improve the learning and memory ability of SAH rats and protect brain nerve function,and its mechanism may be related to the activation of autophagy-related protein expression in hippocampus of SAH rats.
引文
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[12]卢志刚,黄家彬,刘芸,等.醒脑静注射液对急性脑梗死Keap1-Nrf2/ARE氧化应激通路的影响[J].广东医学,2016,37(20):3127-3129.
[13]翟娜,刘岩.醒脑静注射液对高血压脑出血患者炎症因子的影响[J].中国实用神经疾病杂志,2015,18(8):118-119.
[14]吴作林,陈健,郭飞,等.醒脑静对老年脑梗死患者神经细胞修复相关细胞因子的影响[J].医学综述,2016,22(21):4312-4315.
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[16]Komatsu M. Liver autophagy:physiology and pathology[J]. J Biochem,2012,152(1):5-15.
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[19]刘俊杰,赵雅宁,刘仁杰,等.细胞外调节蛋白激酶信号通路对蛛网膜下腔出血大鼠神经元自噬及早期脑损伤的影响[J].中国康复理论与实践,2016,22(10):1121-1126.
[2]赵雅宁,赵旭,郭向飞,等. BQ-123对蛛网膜下腔出血大鼠神经功能缺陷的改善作用及其机制[J].吉林大学学报:医学版,2016,42(5):925-931.
[3]刘振刚,高建亮,孙林林,等.颈内动脉刺破法制作大鼠蛛网膜下腔出血模型[J].中国比较医学杂志,2017,27(6):37-45.
[4]孙林林,刘振刚,张志勇,等.盐酸法舒地尔对蛛网膜下腔出血大鼠学习记忆功能及海马CA1区自噬的影响[J].中国康复理论与实践,2017,23(3):257-262.
[5]朱育军.醒脑静注射液治疗脑出血34例的疗效评价[J].中国药业,2013,22(10):118-119.
[6]李俊芳.醒脑静注射液的药理分析研究[J].中国药物经济学,2014,9(1):48-49.
[7]朱强.醒脑静注射液治疗颅脑外伤昏迷24例[J].河南中医,2015,35(9):2099-2100.
[8]马征,孙雅菲,康玲玲,等.醒脑静注射液治疗急性脑出血患者临床疗效及对血清炎症因子、氧化应激的影响[J].中国实验方剂学杂志,2017,23(19):186-190.
[9]吴伟,高恒.醒脑静注射液联合尼莫地平改善颅内动脉瘤夹闭术后脑血管痉挛25例[J].中国药业,2015,24(20):104-105.
[10]张继方,王厚中,刘伟,等.醒脑静注射液治疗动脉瘤性蛛网膜下腔出血认知功能损害的疗效评价[J].中西医结合心脑血管病杂志,2015,13(1):46-48.
[11]马斌,刘璐,张扬,等.醒脑静注射液对高血压脑梗死大鼠模型的神经保护作用[J].中西医结合心脑血管病杂志,2014,12(2):212-214.
[12]卢志刚,黄家彬,刘芸,等.醒脑静注射液对急性脑梗死Keap1-Nrf2/ARE氧化应激通路的影响[J].广东医学,2016,37(20):3127-3129.
[13]翟娜,刘岩.醒脑静注射液对高血压脑出血患者炎症因子的影响[J].中国实用神经疾病杂志,2015,18(8):118-119.
[14]吴作林,陈健,郭飞,等.醒脑静对老年脑梗死患者神经细胞修复相关细胞因子的影响[J].医学综述,2016,22(21):4312-4315.
[15]Levine B,Mizushima N,Virgin HW. Autophagy in immunity and inflammation[J]. Nature,2011,469(7330):323-325.
[16]Komatsu M. Liver autophagy:physiology and pathology[J]. J Biochem,2012,152(1):5-15.
[17]Codogno P,Meijer A. Autophagy and signaling:their role in cell survival and cell death[J]. Cell Death Differ,2005,12(2):1509-1518.
[18]Shintani T,Klionsky D. Autophagy in health and disease:a double-edged sword[J]. Science,2004,306(5698):990-995.
[19]刘俊杰,赵雅宁,刘仁杰,等.细胞外调节蛋白激酶信号通路对蛛网膜下腔出血大鼠神经元自噬及早期脑损伤的影响[J].中国康复理论与实践,2016,22(10):1121-1126.