加味升降散对血管性痴呆模型大鼠海马神经元表达的影响
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摘要
目的:探讨加味升降散对血管性痴呆(VD)模型大鼠认知障碍的改善效果及其相关作用机制。方法:将90只SPF级SD大鼠随机分为假手术组,模型组,高、中、低剂量组(14、7、3.5mL/kg),石杉碱甲组(0.1mg/kg),每组15只。采用双侧颈总动脉永久结扎法制备VD模型。剔除模型不成功的大鼠,每组随机选取12只作为研究对象。连续灌胃28d,每天3次。应用Morris水迷宫检测各组大鼠学习记忆力;尼氏染色观察海马组织变化;ELISA检测大鼠海马和结肠中SP、VIP的含量;Western Blot检测大鼠海马中Bcl-2、Bax和Caspase-3蛋白表达情况。结果:与模型组比较,各给药组可改善大鼠的定位航行能力与学习记忆能力(P<0.05)。尼氏染色结果表明,各给药组可修复VD模型大鼠海马损伤。各给药组大鼠海马中SP、VIP含量均高于模型组(P<0.05);高、中、低剂量组大鼠结肠中SP、VIP含量均高于模型组(P<0.05)。各给药组大鼠海马中Bcl-2蛋白表达均高于模型组(P<0.05),而Bax和Caspase-3蛋白的表达低于模型组(P<0.05)。结论:加味升降散能够改善VD模型大鼠大脑认知水平。其作用机制可能是通过调节大鼠胃肠功能来改善VD大鼠大脑认知水平,通过上调大鼠海马中Bcl-2蛋白水平,下调Bax和Caspase-3蛋白水平,改善VD模型大鼠海马神经元凋亡状况。
Objective: To investigate the effects and mechanism of Modified Shengjiang Powder on cognitive disorder in vascular dementia(VD) model rats. Methods: Ninety SPF rats were randomly divided into sham operation group, model group,high dose group(14 mL/kg), middle dose group(7 mL/kg), low dose group(3.5 mL/kg), and huperzine A group(0.1 mg/kg), with15 rats in each group. The model of VD was established by 2-VO technique. The unsuccessful modeling rats were excluded and12 rats were randomly selected from each group. Continuous gavage for 28 d, 3 times a day. The ability of learning and memory of rats were evaluated by Morris water maze. Nissl staining method was used to observe the hippocampal tissue changes. The contents of SP, VIP in the hippocampus and colon of rats were detected by ELISA. The expression of Bcl-2, Bax and Caspase-3 protein in hippocampus were detected by Western Blot. Results: Compared with the model group, the rats in the administration group improved the ability of learning and memory, as well as the ability of locating navigation(P<0.05). Nissle staining result showed that treatment group could repair the hippocampal injury of VD model rats. The contents of SP, VIP in hippocampus of rats in treatment group were higher than those in model group(P<0.05). The expression of SP, VIP in the colon of rats in high,middle and low dose groups were higher than those in the model group(P<0.05). The protein level of Bcl-2 in treatment group was higher than in the model group(P<0.05), while the protein levels of Bax and Caspase-3 were significantly lower than that in the model group(P<0.05). Conclusion: Modified Shengjiang Powder could improve the cognitive level of VD model rats. The mechanism may be to improve the cognitive level of VD rats by regulating the gastrointestinal function of rats, and up-regulate the level of Bcl-2 protein in the hippocampus of rats, down-regulate the levels of bax and Caspase-3, and improve the apoptotic state of hippocampal neurons.
Objective: To investigate the effects and mechanism of Modified Shengjiang Powder on cognitive disorder in vascular dementia(VD) model rats. Methods: Ninety SPF rats were randomly divided into sham operation group, model group,high dose group(14 mL/kg), middle dose group(7 mL/kg), low dose group(3.5 mL/kg), and huperzine A group(0.1 mg/kg), with15 rats in each group. The model of VD was established by 2-VO technique. The unsuccessful modeling rats were excluded and12 rats were randomly selected from each group. Continuous gavage for 28 d, 3 times a day. The ability of learning and memory of rats were evaluated by Morris water maze. Nissl staining method was used to observe the hippocampal tissue changes. The contents of SP, VIP in the hippocampus and colon of rats were detected by ELISA. The expression of Bcl-2, Bax and Caspase-3 protein in hippocampus were detected by Western Blot. Results: Compared with the model group, the rats in the administration group improved the ability of learning and memory, as well as the ability of locating navigation(P<0.05). Nissle staining result showed that treatment group could repair the hippocampal injury of VD model rats. The contents of SP, VIP in hippocampus of rats in treatment group were higher than those in model group(P<0.05). The expression of SP, VIP in the colon of rats in high,middle and low dose groups were higher than those in the model group(P<0.05). The protein level of Bcl-2 in treatment group was higher than in the model group(P<0.05), while the protein levels of Bax and Caspase-3 were significantly lower than that in the model group(P<0.05). Conclusion: Modified Shengjiang Powder could improve the cognitive level of VD model rats. The mechanism may be to improve the cognitive level of VD rats by regulating the gastrointestinal function of rats, and up-regulate the level of Bcl-2 protein in the hippocampus of rats, down-regulate the levels of bax and Caspase-3, and improve the apoptotic state of hippocampal neurons.
引文
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[6]刘忠锦,张海燕,刘佳俊.石杉碱甲对血管性痴呆大鼠胶质细胞源性神经营养因子的表达及氧化应激水平的影响.中国生化药物杂志,2014,34(6):44-46,50
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[12]胡跃强,赖菁菁,汪庭龙,等.温肺降浊方对血管性痴呆大鼠海马神经元凋亡的影响.中华中医药杂志,2017,32(4):1804-1807
[13]Theofilas P,Bedner P,Huttmann K,et al.The proapoptotic Bcl-2 homology domain 3-only protein Bim is not critical for acute excitotoxic celldeath.J Neuropathol Exp Neurol,2009,68(1):102-110
[14]刘伟国,郑学胜,杨小锋,等.Bcl-2融合蛋白对颅脑损伤细胞凋亡的影响.中华神经外科杂志,2006,22(2):86-88
[15]张暄琳,李毅,刘丽,等.炎症因子对人颗粒细胞凋亡调控基因P53、Bax、Casepase-3、Bcl-2、Servivin表达的影响.实用妇产科杂志,2016,32(7):519-522
[2]孙晓彩,李力,张敏,等.Wistar大鼠海马CA1区、CA3区和齿状回区的解剖分割.中国应用生理学杂志,2012,28(2):189-192
[3]LongstrethWT,ArnoldJA,BeauchampNJ,etal.Incidence,manifestations and predictors of worsening white matter on serial cranial magnetic resonance imaging in the elderly:The cardiovascular health study.Stroke,2005,36(1):56-61
[4]Reinhold S,Stefan R,JoséF,et al.Diffusion-weighted imaging and cognition in the leuko-ariosis and disability in the elderly study.Stroke,2010,41(5):402-408
[5]张英菊.当归对脑出血大鼠神经细胞凋亡及相关蛋白表达的影响.兰州:甘肃中医药大学,2015
[6]刘忠锦,张海燕,刘佳俊.石杉碱甲对血管性痴呆大鼠胶质细胞源性神经营养因子的表达及氧化应激水平的影响.中国生化药物杂志,2014,34(6):44-46,50
[7]于涛,赵利娜,陈其奎.脑肠肽与功能性消化不良.现代消化及介入诊疗,2012,17(4):241-245
[8]陈求招.功能性消化不良与胃粘膜胃动素、降钙素基因相关肽的关系.福州:福建医科大学,2012
[9]王韶轩.脑肠肽与消化及神经系统基础与临床.济南:山东大学出版社,2010:54-59
[10]路长林.脑肠肽基础与临床.上海:第二军医大学出版社,2000:29-33
[11]Sitailo L A,Jerome-Morais A,Denning M F.Mcl-1 functions as major epidermal survival protein required for proper keratinocyte differentiation.J Invest Dermatol,2009,129(6):1351-1360
[12]胡跃强,赖菁菁,汪庭龙,等.温肺降浊方对血管性痴呆大鼠海马神经元凋亡的影响.中华中医药杂志,2017,32(4):1804-1807
[13]Theofilas P,Bedner P,Huttmann K,et al.The proapoptotic Bcl-2 homology domain 3-only protein Bim is not critical for acute excitotoxic celldeath.J Neuropathol Exp Neurol,2009,68(1):102-110
[14]刘伟国,郑学胜,杨小锋,等.Bcl-2融合蛋白对颅脑损伤细胞凋亡的影响.中华神经外科杂志,2006,22(2):86-88
[15]张暄琳,李毅,刘丽,等.炎症因子对人颗粒细胞凋亡调控基因P53、Bax、Casepase-3、Bcl-2、Servivin表达的影响.实用妇产科杂志,2016,32(7):519-522