黄芪注射液抑制糖尿病大鼠海马神经元凋亡的机制研究
|
摘要
[目的]探讨黄芪注射液对糖尿病模型大鼠认知功能的影响及其作用机制。[方法]40只雄性SD大鼠,除空白组(n=10)外腹腔注射链脲佐菌素(60 mg/kg)复制Ⅰ型糖尿病大鼠模型,随机分为模型组和黄芪注射液组(1.6 mg/kg),给药10周。以Morris水迷宫法进行学习记忆功能测试,随即处死大鼠分离血清及脑组织,分别测定空腹血糖、空腹胰岛素水平和乙酰胆碱酯酶含量,RT-PCR法测定海马Bcl-2及Bax mRNA表达情况,免疫蛋白印迹法(Western blot)测定海马乙酰胆碱酯酶和Caspase-3的蛋白表达及免疫组化法观察海马各区Caspase-3蛋白的阳性表达情况。[结果]黄芪注射液能显著提高糖尿病脑病大鼠学习记忆能力,降低糖尿病大鼠空腹血糖水平及AchE含量,升高血清胰岛素水平,减少海马AchE及Caspase-3蛋白的表达,升高海马抗凋亡基因Bcl-2的表达,降低促凋亡基因Bax的表达。[结论]黄芪注射液能够显著改善糖尿病大鼠学习记忆功能,其作用机制可能与减少AchE含量、抑制神经细胞凋亡有关。
[Objective] To explore the effects of Huangqi injection on learning and memory abilities in diabetic rats and its mechanism research. [Methods] Forty male SD rats, except the blank group(n =10),were intraperitoneal injected Streptozotocin(60 mg/kg) to establish type 1 diabetic rats, the modeled rats were equally and randomly divided into the model group and Huangqi injection group. All drugs were administrated for 10 weeks. The behavior test were performed on the Morris water maze method, and then the rats were executed for blood and brain sampling. Blood glucose, insulin and acetylcholinesterase level were detected. The expression of Bcl-2 and Bax in the hippocampus was examined by quantitative real-time PCR. The method of Western blotting was used to measure the expression of AchE. The expression of Caspase-3 in the hippocampus nerve cells was examined immunohistochemistry. [Results] Huangqi injection obviously improved the learning and memory abilities of diabetic encephalopathy rats, reduced blood glucose and AchE level and incresed the insulin level. The expression of AchE and Caspase-3 protein were obviously reduced in the hippocampus of rats. The antiapoptosis gene Bcl-2 mRNA epression was increased while the apoptosis Bax mRNA expression was decreased in hippocampal area in Huangqi injection group. [Conclusion] Huangqi Injection obviously improved the learning and memory abilities of diabetic rats, its mechanism may be related to reduce AchE level, inhibit neuronal apoptosis.
[Objective] To explore the effects of Huangqi injection on learning and memory abilities in diabetic rats and its mechanism research. [Methods] Forty male SD rats, except the blank group(n =10),were intraperitoneal injected Streptozotocin(60 mg/kg) to establish type 1 diabetic rats, the modeled rats were equally and randomly divided into the model group and Huangqi injection group. All drugs were administrated for 10 weeks. The behavior test were performed on the Morris water maze method, and then the rats were executed for blood and brain sampling. Blood glucose, insulin and acetylcholinesterase level were detected. The expression of Bcl-2 and Bax in the hippocampus was examined by quantitative real-time PCR. The method of Western blotting was used to measure the expression of AchE. The expression of Caspase-3 in the hippocampus nerve cells was examined immunohistochemistry. [Results] Huangqi injection obviously improved the learning and memory abilities of diabetic encephalopathy rats, reduced blood glucose and AchE level and incresed the insulin level. The expression of AchE and Caspase-3 protein were obviously reduced in the hippocampus of rats. The antiapoptosis gene Bcl-2 mRNA epression was increased while the apoptosis Bax mRNA expression was decreased in hippocampal area in Huangqi injection group. [Conclusion] Huangqi Injection obviously improved the learning and memory abilities of diabetic rats, its mechanism may be related to reduce AchE level, inhibit neuronal apoptosis.
引文
[1]Cukierman T,Gerstein HC,Williamson JD,et al.Cognitive decline and dementia in diabetes-systematic overview of prospective observational studies[J].Diabetologia,2005,48(12):2460-2469.
[2]Rawal N,Corti O,Sacchetti P,et al.Parkin protects dopaminergic neurons from excessive Wnt/β-catenin signaling[J].Biochem Biophys Res Commun,2009,23,388(3):473-478.
[3]Caricasole A,Copani A,Caraci F,et al.Induction of Dickkopf-1,a negative modulator of the wnt pathway,is Associated with neuronal degeneration in alzheimer’s brain[J].J Neurosci,2004,24(26):6021-6027.
[4]Sadeghi A,Hami J,Razavi S,et al.The effect of diabetes mellitus on apoptosis in hippocampus:cellular and molecular aspects[J].Int J Prev Med,2016,7:57.
[5]罗焕敏.海马结构──从形态、功能到可塑性、衰老性变化[J].神经解剖学杂志,1996,12(2):177-184.
[6]Zhan Y,van de Water B,Wang Y,et al.The roles of caspase-3 and bcl-2 in chemically-induced apoptosis but not necrosis of renal epithelial cells[J].Oncogene,1999,11,18(47):6505-6512.
[7]Venero JL,Burguillos MA,Joseph B,et al.Caspases Playing in the Field of Neuroinflammation:Old and New Players[J].Dev Neurosci,2013,35(2-3):88-101.
[8]Dorszewska J.Cell biology of normal brain aging:synaptic plasticitycell death[J].Aging Clin Exp Res,2013,25(1):25-34.
[9]杨磊.乙酰胆碱酯酶与细胞凋亡的相关性的研究[D].上海:中国科学院研究生院(上海生命科学研究院),2003.
[10]胡涛,刘希伟.脑缺血对免疫系统乙酰胆碱酯酶及Caspase-3表达的影响[J].中国免疫学杂志,2008,24(5):425-429.
[11]王晓华,何杏容,华红艳,等.丹参联合黄芪对糖尿病肾病大鼠肾脏线粒体呼吸及能量代谢的影响[J].临床与病理杂志,2015,35(2):243-247.
[12]赵胜利,高建玲,翟丽娜.丹参及黄芪注射液对心脑血管病患者血液流变学的影响[J].实用中医药杂志,2002,18(6):5.
[13]万朋,王师,郎茂林,等.黄芪对阿尔茨海默病模型大鼠学习记忆能力的影响[J].吉林医药学院学报,2011,32(2):69-73.
[14]张雅丽,高维娟,闫凤霞,等.黄芪注射液抑制缺氧缺糖后复氧复糖大鼠海马神经细胞凋亡的研究[J].中国老年学杂志,2009,29(7):793-796.
[15]闫凤霞,高维娟,钱涛,等.黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元caspase-3表达的作用[J].中国药理学通报,2011,27(1):85-90.
[16]焦俊霞,高维娟,钱涛,等.黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元bcl-2、bax及cyt-c蛋白形态学表达的影响[J].中国老年学杂志,2011,31(3):445-448.
[17]闫凤霞,高维娟,钱涛,等.黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元凋亡的自由基机制[J].中国老年学杂志,2011,31(8):1347-1351.
[2]Rawal N,Corti O,Sacchetti P,et al.Parkin protects dopaminergic neurons from excessive Wnt/β-catenin signaling[J].Biochem Biophys Res Commun,2009,23,388(3):473-478.
[3]Caricasole A,Copani A,Caraci F,et al.Induction of Dickkopf-1,a negative modulator of the wnt pathway,is Associated with neuronal degeneration in alzheimer’s brain[J].J Neurosci,2004,24(26):6021-6027.
[4]Sadeghi A,Hami J,Razavi S,et al.The effect of diabetes mellitus on apoptosis in hippocampus:cellular and molecular aspects[J].Int J Prev Med,2016,7:57.
[5]罗焕敏.海马结构──从形态、功能到可塑性、衰老性变化[J].神经解剖学杂志,1996,12(2):177-184.
[6]Zhan Y,van de Water B,Wang Y,et al.The roles of caspase-3 and bcl-2 in chemically-induced apoptosis but not necrosis of renal epithelial cells[J].Oncogene,1999,11,18(47):6505-6512.
[7]Venero JL,Burguillos MA,Joseph B,et al.Caspases Playing in the Field of Neuroinflammation:Old and New Players[J].Dev Neurosci,2013,35(2-3):88-101.
[8]Dorszewska J.Cell biology of normal brain aging:synaptic plasticitycell death[J].Aging Clin Exp Res,2013,25(1):25-34.
[9]杨磊.乙酰胆碱酯酶与细胞凋亡的相关性的研究[D].上海:中国科学院研究生院(上海生命科学研究院),2003.
[10]胡涛,刘希伟.脑缺血对免疫系统乙酰胆碱酯酶及Caspase-3表达的影响[J].中国免疫学杂志,2008,24(5):425-429.
[11]王晓华,何杏容,华红艳,等.丹参联合黄芪对糖尿病肾病大鼠肾脏线粒体呼吸及能量代谢的影响[J].临床与病理杂志,2015,35(2):243-247.
[12]赵胜利,高建玲,翟丽娜.丹参及黄芪注射液对心脑血管病患者血液流变学的影响[J].实用中医药杂志,2002,18(6):5.
[13]万朋,王师,郎茂林,等.黄芪对阿尔茨海默病模型大鼠学习记忆能力的影响[J].吉林医药学院学报,2011,32(2):69-73.
[14]张雅丽,高维娟,闫凤霞,等.黄芪注射液抑制缺氧缺糖后复氧复糖大鼠海马神经细胞凋亡的研究[J].中国老年学杂志,2009,29(7):793-796.
[15]闫凤霞,高维娟,钱涛,等.黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元caspase-3表达的作用[J].中国药理学通报,2011,27(1):85-90.
[16]焦俊霞,高维娟,钱涛,等.黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元bcl-2、bax及cyt-c蛋白形态学表达的影响[J].中国老年学杂志,2011,31(3):445-448.
[17]闫凤霞,高维娟,钱涛,等.黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元凋亡的自由基机制[J].中国老年学杂志,2011,31(8):1347-1351.