大黄酚对小鼠脑缺血再灌注损伤沉默信息调节因子3和水通道蛋白4表达的影响
摘要
目的测定脑组织中沉默信息调节因子(SIRT)3和水通道蛋白(AQP)4的表达,探讨大黄酚对脑缺血再灌注损伤的神经保护作用。方法随机将昆明小鼠分为空白对照组、假手术组、模型组、大黄酚组。采用改良的Himori法暂时性阻断双侧颈总动脉建立脑缺血再灌注模型。观察脑缺血5 min再灌注24 h后小鼠的行为学表现,计算神经功能损伤评分,应用免疫组化法测定脑组织中沉默信息调节因子(SIRT)3,水通道蛋白(AQP)4的表达,测定脑组织中的超氧化物歧化酶(SOD)活力。结果大黄酚能明显改善脑缺血再灌注后小鼠的神经功能损伤情况;大黄酚可提高脑组织中SOD的活力;小鼠脑缺血再灌注后24 h SIRT3及AQP4表达均增强,而应用大黄酚可以降低AQP4以及上调SIRT3的表达。SIRT3的表达与SOD活力呈正相关。结论大黄酚是通过上调SIRT3的表达,增强抗细胞凋亡及SOD的抗氧化作用;下调AQP4的表达,减轻脑缺血后脑水肿的发生,发挥其神经保护作用。
引文
1 Dorado L,Millán M,Dávalos A.Reperfusion therapies for acute ischemic stroke:an update[J].Curr Cardiol Rev,2014;10(4):327-35.
2 Gilgun-Sherki Y,Rosenbaum Z,Melamed E,et al.Antioxidant therapy in acute central nervous system injury:current state[J].Pharmacol Rev,2002;54(2):271-84.
3 李沙,黄艳秋,游红琴,等.脑缺血再灌注损伤模型大鼠炎性反应相关信号研究[J].中国神经免疫学和神经病学杂志,2012;19(6):453-61.
4 Harrison DC,Davis RP,Bond BC,et al.Caspase mRNA expression in a rat model of focal cerebral ischemia [J].Brain Res Mol Brain Res,2001;89(1-2):133-46.
5 Wang Q,Kalogeris TJ,Wang M,et al.Antecedent ethanol attenuates cerebral ischemia/reperfusion-induced leukocyte-endothelial adhensive interactions and delayed neuronal death:role of large conductance,Ca2+-activated K+ channels[J].Microcirculation,2010;17(6):427-38.
6 Martin LJ.Neuronal cell death in nervous system development and injury (review) [J].Int J Mol Med,2011;7(5):455-78.
7 Himori N,Watanabe H,Nobuhide A,et al.Cerebral ischemia model with conscious mice:involvement of NMDA receptor activation and derangement of learning and memory ability [J].J Pharmacol Meth,1990;23(4):311-27.
8 Garecia JH,Wagner S,Liu KF,et al.Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats.Statistical validation [J].Stroke,1995;26(4):627-34.
9 王树,张力,张丹参,等.大黄酚对脑缺血再灌注小鼠探索功能的影响[J].时珍国医国药,2007;18(12):3011-3.
10 王树,薛贵平,张丹参,等.大黄酚对脑缺血再灌注小鼠学习记忆障碍及耐缺氧的影响[J].陕西医学杂志,2008;37(4):402-4.
11 王树,张丹参,张力,等.大黄酚对脑缺血再灌注小鼠记忆功能的保护作用[J].中国老年学杂志,2009;29(15):1934-6.
12 颜娟.大黄酚脂质体对小鼠脑缺血再灌注损伤的神经保护作用及机制研究[D].石家庄:河北医科大学,2014.
13 Ribeiro MC,Hirt L,Bogousslavsky J,et al.Time course of Aquaporin expression after transient focal cerebral ischemia in mice[J].Neurossci Res,2006;83(7):1231-40.
14 Arge P,Preston GM.Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons:member of an ancient channel family[J].Proc Acad Sci USA,1991;88(24):11110-4.
15 Zeng XN,Xie LL,Liang R,et al.AQP4 knockout aggravates ischemia/reperfusion injury in mice[J].CNS Neurosci Therapeut,2012;18(5):388-94.
16 Thrane AS,Rappold PM,Fujita T,et al.Critical role of aquaporin-4(AQP4) in astrocytic Ca2+ signaling events elicited by cerebral edema [J].Proc Natl Acad Sci USA,2011;108(2):846-51.
17 刘青.大黄总蒽醌及其衍生物对LoVo、NRK细胞AQP2、AQP4表达的调节效应与机制[D].西安:第四军医大学,2009.
18 Qiu X,Brown K,Hirschey MD,et al.Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation[J].Cell Metab,2010;12(6):662-7.
19 Porter GA,Urciuoli WR,Brookes PS,et al.SIRT3 deficiency exacerbates ischemia-reperfusion injury:implication for aged hearts[J].Am J Physiol Heart Circ Physiol,2014;306(12):H1602-9.
2 Gilgun-Sherki Y,Rosenbaum Z,Melamed E,et al.Antioxidant therapy in acute central nervous system injury:current state[J].Pharmacol Rev,2002;54(2):271-84.
3 李沙,黄艳秋,游红琴,等.脑缺血再灌注损伤模型大鼠炎性反应相关信号研究[J].中国神经免疫学和神经病学杂志,2012;19(6):453-61.
4 Harrison DC,Davis RP,Bond BC,et al.Caspase mRNA expression in a rat model of focal cerebral ischemia [J].Brain Res Mol Brain Res,2001;89(1-2):133-46.
5 Wang Q,Kalogeris TJ,Wang M,et al.Antecedent ethanol attenuates cerebral ischemia/reperfusion-induced leukocyte-endothelial adhensive interactions and delayed neuronal death:role of large conductance,Ca2+-activated K+ channels[J].Microcirculation,2010;17(6):427-38.
6 Martin LJ.Neuronal cell death in nervous system development and injury (review) [J].Int J Mol Med,2011;7(5):455-78.
7 Himori N,Watanabe H,Nobuhide A,et al.Cerebral ischemia model with conscious mice:involvement of NMDA receptor activation and derangement of learning and memory ability [J].J Pharmacol Meth,1990;23(4):311-27.
8 Garecia JH,Wagner S,Liu KF,et al.Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats.Statistical validation [J].Stroke,1995;26(4):627-34.
9 王树,张力,张丹参,等.大黄酚对脑缺血再灌注小鼠探索功能的影响[J].时珍国医国药,2007;18(12):3011-3.
10 王树,薛贵平,张丹参,等.大黄酚对脑缺血再灌注小鼠学习记忆障碍及耐缺氧的影响[J].陕西医学杂志,2008;37(4):402-4.
11 王树,张丹参,张力,等.大黄酚对脑缺血再灌注小鼠记忆功能的保护作用[J].中国老年学杂志,2009;29(15):1934-6.
12 颜娟.大黄酚脂质体对小鼠脑缺血再灌注损伤的神经保护作用及机制研究[D].石家庄:河北医科大学,2014.
13 Ribeiro MC,Hirt L,Bogousslavsky J,et al.Time course of Aquaporin expression after transient focal cerebral ischemia in mice[J].Neurossci Res,2006;83(7):1231-40.
14 Arge P,Preston GM.Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons:member of an ancient channel family[J].Proc Acad Sci USA,1991;88(24):11110-4.
15 Zeng XN,Xie LL,Liang R,et al.AQP4 knockout aggravates ischemia/reperfusion injury in mice[J].CNS Neurosci Therapeut,2012;18(5):388-94.
16 Thrane AS,Rappold PM,Fujita T,et al.Critical role of aquaporin-4(AQP4) in astrocytic Ca2+ signaling events elicited by cerebral edema [J].Proc Natl Acad Sci USA,2011;108(2):846-51.
17 刘青.大黄总蒽醌及其衍生物对LoVo、NRK细胞AQP2、AQP4表达的调节效应与机制[D].西安:第四军医大学,2009.
18 Qiu X,Brown K,Hirschey MD,et al.Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation[J].Cell Metab,2010;12(6):662-7.
19 Porter GA,Urciuoli WR,Brookes PS,et al.SIRT3 deficiency exacerbates ischemia-reperfusion injury:implication for aged hearts[J].Am J Physiol Heart Circ Physiol,2014;306(12):H1602-9.