番茄红素对血管性痴呆大鼠的抗凋亡作用
发布时间：2018-09-03 08:56
【摘要】：目的:血管性痴呆(Vascular dementia,VD)是指由于各种脑血管因素(如缺血、出血、急慢性脑供血障碍等)引起脑组织损伤后导致认知功能障碍和记忆力下降的一种临床综合征。缺血性脑血管病在血管性痴呆的发病原因中居首位。近年,随着人口老龄化的进程,脑卒中和心血管疾病的发病率持续上升,血管性痴呆的患病率也随之增高,但其确切的发病机制仍未阐明。目前认为氧化应激、兴奋性氨基酸毒性、炎症反应、细胞凋亡等共同参与血管性痴呆的发生发展。细胞凋亡(Apoptosis)为细胞的程序性死亡,是细胞受到伤害性刺激因素导致细胞死亡的重要机制。细胞凋亡分为外源性途径和内源性途径,外源性凋亡途径,即死亡受体途径,经Fas蛋白与细胞膜上的受体结合并激活,招募位于细胞内的半胱氨酸天冬氨酸蛋白酶(Caspase),启动细胞凋亡;内源性凋亡途径,即线粒体途径,内外刺激使线粒体膜的通透性增强,引起细胞色素C的释放,其与凋亡激活因子1(Apaf-1)形成复合体,启动凋亡程序。Caspase9和caspase3是凋亡途径级联反应链上的关键性生物分子。Caspase3通过切割多聚ADP核糖聚合酶(PARP)等引起DNA损伤,最终导致细胞死亡。番茄红素(Lycopene,LP)是一种高度不饱和的脂溶性色素,广泛存在于番茄、西瓜等蔬菜水果中。番茄红素具有强大的抗氧化能力,能够清除过多的氧自由基,抵抗衰老、增强人体免疫力、减少心脑血管疾病发生、抑制肿瘤生长等。近年来,多项有关番茄红素在脑血管病方面作用的研究表明,番茄红素可通过其强大的抗氧化作用对脑血管起到保护作用。但番茄红素是否可通过抗细胞凋亡发挥脑保护作用鲜有报道。本实验通过结扎大鼠双侧颈总动脉法(2-VO)制备血管性痴呆动物模型,应用番茄红素进行干预,观察番茄红素对血管性痴呆大鼠行为认知功能及海马CA1区凋亡指标caspase9、caspase3表达水平的影响。方法:对2月龄健康雄性Sprague-Dawley(SD)大鼠进行双侧颈总动脉结扎制备血管性痴呆动物模型。假手术组10只,只分离颈部肌肉,暴露血管,不结扎动脉。将手术后存活动物按照痴呆动物标准进行筛选,符合标准的动物随机分为模型组、番茄红素组,每组约10只。番茄红素组大鼠每日给予番茄红素10mg/kg,模型组和假手术组大鼠给予等容量植物油,持续给药6周。干预结束后,行水迷宫实验,测定各组大鼠逃避潜伏期及穿越平台的次数。后留取大鼠海马组织,应用免疫组织化学方法(Immunohistochemistry)和免疫印迹法(Western blot)测定海马CA1区caspase3、caspase9的表达水平。结果:1番茄红素对血管性痴呆大鼠的行为学影响水迷宫定位航行实验结果表明,三组大鼠的逃避潜伏期从第2天开始均较前1天缩短,表明三组大鼠都有一定的学习能力,但同时三组大鼠之间的学习能力又有一定的差别,三组大鼠之间的逃避潜伏期有显著差异(P0.05)。模型组与假手术组相比,前4天的逃避潜伏期明显延长(P0.05);番茄红素组与模型组相比,前4天的逃避潜伏期明显缩短(P0.05);番茄红素组与假手术组相比,前4天的逃避潜伏期延长(P0.05)。第5天进行穿越平台次数实验,穿越平台次数的多少与大鼠的记忆功能有关,穿越平台次数越多说明大鼠的记忆能力越好。模型组与假手术组相比,穿越平台次数明显减少(P0.05);番茄红素组与模型组相比,穿越平台次数明显增多(P0.05);番茄红素组与假手术组相比,穿越平台次数减少,但无显著统计学差异(P0.05)。2海马CA1区caspase3、caspase9表达水平western blot结果大鼠海马CA1区caspase3、caspase9表达情况:模型组与假手术组相比,大鼠海马CA1区caspase3、caspase9蛋白表达水平明显增加(P0.05);番茄红素组与模型组相比,海马CA1区caspase3、caspase9蛋白表达水平明显减少(P0.05);番茄红素组与假手术组相比,凋亡蛋白表达水平增加,但无明显统计学差异(P0.05)。3海马CA1区caspase3免疫组织化学结果大鼠海马CA1区caspase3蛋白的免疫组化结果:模型组与假手术组相比,海马神经元萎缩,caspase3蛋白表达阳性细胞数明显增加(P0.05);番茄红素组与模型组相比,caspase3蛋白表达阳性细胞数明显减少(P0.05);番茄红素组与假手术组相比,caspase3蛋白表达阳性细胞数明显增加(P0.05)。结论:1番茄红素能够明显缩短血管性痴呆大鼠的逃避潜伏期,使其穿越平台次数明显增加,表明番茄红素可提高血管性痴呆大鼠的学习记忆能力。2番茄红素可以下调血管性痴呆大鼠海马组织CA1区caspase3、caspase9的表达水平,提示番茄红素可能通过抗细胞凋亡对海马神经元起到保护作用。
[Abstract]:Objective: Vascular dementia (VD) is a kind of clinical syndrome that causes cognitive impairment and memory loss after brain tissue injury caused by various cerebrovascular factors (such as ischemia, hemorrhage, acute or chronic cerebral blood supply disorders, etc.). Ischemic cerebrovascular disease is the most common cause of vascular dementia. In recent years, with the development of human beings, the incidence of VD is increasing. As the aging of the mouth progresses, the incidence of stroke and cardiovascular disease continues to rise, and the incidence of vascular dementia increases, but the exact pathogenesis remains unclear. S) Procedural cell death is an important mechanism of cell death induced by noxious stimuli. Apoptosis is divided into exogenous and endogenous pathways, and exogenous pathways of apoptosis, i.e. death receptor pathway, are activated by Fas protein binding to receptors on cell membrane to recruit cysteine aspartate methionine in cells. Caspase initiates apoptosis; endogenous apoptosis pathway, i.e. mitochondrial pathway, enhances the permeability of mitochondrial membrane and induces the release of cytochrome C. Caspase 9 and caspase 3 are key biological molecules in the cascade reaction chain of apoptosis pathway. Lycopene (LP) is a highly unsaturated lipid-soluble pigment, widely found in tomatoes, watermelons and other vegetables and fruits. Lycopene has a strong antioxidant capacity, can scavenge excessive oxygen free radicals, resist aging, increase. In recent years, a number of studies on the role of lycopene in cerebrovascular diseases have shown that lycopene can protect cerebrovascular through its strong antioxidant effect. But whether lycopene can play a fresh role in brain protection by anti-apoptosis In this study, vascular dementia animal model was established by ligating bilateral common carotid arteries (2-VO) in rats. Lycopene was used to interfere with the behavior and cognitive function of vascular dementia rats and the expression of apoptotic markers caspase 9 and caspase 3 in hippocampal CA1 region. Methods: Sprague-Dawley (2-month-old male) was used to observe the effect of lycopene on the behavior and cognitive function of vascular dementia rats. SD) Rats were ligated bilateral common carotid arteries to establish vascular dementia animal model. In sham operation group, 10 rats were isolated neck muscles, exposed blood vessels, and not ligated arteries. The rats in the model group and sham operation group were given the same volume of vegetable oil for 6 weeks. After the intervention, the escape latency and the number of crossing platforms were measured by water maze test. The hippocampus tissues of the rats were harvested by immunohistochemistry and Western blotting. The expression levels of caspase 3 and caspase 9 in hippocampal CA1 region were determined by tern blot. Results: 1. Lycopene affected the behavior of vascular dementia rats. The results of water maze navigation experiment showed that the escape latency of three groups of rats was shorter from the second day than the first day, indicating that the three groups of rats had certain learning ability, but at the same time, the three groups of rats had certain learning ability. The escape latency of model group was significantly longer than that of sham operation group (P 0.05); the escape latency of lycopene group was significantly shorter than that of model group (P 0.05); the escape latency of lycopene group was significantly shorter than that of sham operation group (P 0.05); and the escape latency of lycopene group was significantly shorter than that of sham operation group (P 0.05). The escape latency was prolonged (P 0.05). The number of times crossing the platform was related to the memory function of rats. The more times crossing the platform, the better the memory ability of rats. The expression of caspase 3 and caspase 9 in hippocampal CA1 region was detected by Western blot. Compared with sham operation group, the expression of caspase 3 and caspase 9 in hippocampal CA1 region of rats in model group was lower than that in sham operation group. The expression of Caspase-3 and caspase-9 protein in hippocampal CA1 region was significantly lower in lycopene group than in model group (P 0.05). The expression of apoptotic protein in lycopene group was significantly higher than that in sham operation group, but there was no significant difference (P 0.05). Immunohistochemical results of caspase-3 protein in CA1 region of horse: Compared with sham-operated group, the number of caspase-3 protein positive cells in hippocampal neurons atrophied and increased significantly in model group (P 0.05); compared with model group, the number of caspase-3 protein positive cells in lycopene group decreased significantly (P 0.05); compared with sham-operated group, the expression of caspase-3 protein in lycopene group decreased significantly (P 0.05); and compared with sham-operated group, the CONCLUSION: Lycopene can significantly shorten the escape latency and increase the number of platforms crossing in vascular dementia rats, indicating that Lycopene can improve the learning and memory ability of vascular dementia rats. 2 Lycopene can down-regulate caspase 3 and caspase 3 in hippocampal CA1 region of vascular dementia rats. The expression level of E9 suggested that lycopene might play a protective role in hippocampal neurons through anti apoptosis.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.13
[Abstract]:Objective: Vascular dementia (VD) is a kind of clinical syndrome that causes cognitive impairment and memory loss after brain tissue injury caused by various cerebrovascular factors (such as ischemia, hemorrhage, acute or chronic cerebral blood supply disorders, etc.). Ischemic cerebrovascular disease is the most common cause of vascular dementia. In recent years, with the development of human beings, the incidence of VD is increasing. As the aging of the mouth progresses, the incidence of stroke and cardiovascular disease continues to rise, and the incidence of vascular dementia increases, but the exact pathogenesis remains unclear. S) Procedural cell death is an important mechanism of cell death induced by noxious stimuli. Apoptosis is divided into exogenous and endogenous pathways, and exogenous pathways of apoptosis, i.e. death receptor pathway, are activated by Fas protein binding to receptors on cell membrane to recruit cysteine aspartate methionine in cells. Caspase initiates apoptosis; endogenous apoptosis pathway, i.e. mitochondrial pathway, enhances the permeability of mitochondrial membrane and induces the release of cytochrome C. Caspase 9 and caspase 3 are key biological molecules in the cascade reaction chain of apoptosis pathway. Lycopene (LP) is a highly unsaturated lipid-soluble pigment, widely found in tomatoes, watermelons and other vegetables and fruits. Lycopene has a strong antioxidant capacity, can scavenge excessive oxygen free radicals, resist aging, increase. In recent years, a number of studies on the role of lycopene in cerebrovascular diseases have shown that lycopene can protect cerebrovascular through its strong antioxidant effect. But whether lycopene can play a fresh role in brain protection by anti-apoptosis In this study, vascular dementia animal model was established by ligating bilateral common carotid arteries (2-VO) in rats. Lycopene was used to interfere with the behavior and cognitive function of vascular dementia rats and the expression of apoptotic markers caspase 9 and caspase 3 in hippocampal CA1 region. Methods: Sprague-Dawley (2-month-old male) was used to observe the effect of lycopene on the behavior and cognitive function of vascular dementia rats. SD) Rats were ligated bilateral common carotid arteries to establish vascular dementia animal model. In sham operation group, 10 rats were isolated neck muscles, exposed blood vessels, and not ligated arteries. The rats in the model group and sham operation group were given the same volume of vegetable oil for 6 weeks. After the intervention, the escape latency and the number of crossing platforms were measured by water maze test. The hippocampus tissues of the rats were harvested by immunohistochemistry and Western blotting. The expression levels of caspase 3 and caspase 9 in hippocampal CA1 region were determined by tern blot. Results: 1. Lycopene affected the behavior of vascular dementia rats. The results of water maze navigation experiment showed that the escape latency of three groups of rats was shorter from the second day than the first day, indicating that the three groups of rats had certain learning ability, but at the same time, the three groups of rats had certain learning ability. The escape latency of model group was significantly longer than that of sham operation group (P 0.05); the escape latency of lycopene group was significantly shorter than that of model group (P 0.05); the escape latency of lycopene group was significantly shorter than that of sham operation group (P 0.05); and the escape latency of lycopene group was significantly shorter than that of sham operation group (P 0.05). The escape latency was prolonged (P 0.05). The number of times crossing the platform was related to the memory function of rats. The more times crossing the platform, the better the memory ability of rats. The expression of caspase 3 and caspase 9 in hippocampal CA1 region was detected by Western blot. Compared with sham operation group, the expression of caspase 3 and caspase 9 in hippocampal CA1 region of rats in model group was lower than that in sham operation group. The expression of Caspase-3 and caspase-9 protein in hippocampal CA1 region was significantly lower in lycopene group than in model group (P 0.05). The expression of apoptotic protein in lycopene group was significantly higher than that in sham operation group, but there was no significant difference (P 0.05). Immunohistochemical results of caspase-3 protein in CA1 region of horse: Compared with sham-operated group, the number of caspase-3 protein positive cells in hippocampal neurons atrophied and increased significantly in model group (P 0.05); compared with model group, the number of caspase-3 protein positive cells in lycopene group decreased significantly (P 0.05); compared with sham-operated group, the expression of caspase-3 protein in lycopene group decreased significantly (P 0.05); and compared with sham-operated group, the CONCLUSION: Lycopene can significantly shorten the escape latency and increase the number of platforms crossing in vascular dementia rats, indicating that Lycopene can improve the learning and memory ability of vascular dementia rats. 2 Lycopene can down-regulate caspase 3 and caspase 3 in hippocampal CA1 region of vascular dementia rats. The expression level of E9 suggested that lycopene might play a protective role in hippocampal neurons through anti apoptosis.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.13
【参考文献】
相关期刊论文 前6条
1 包华音;;番茄红素药理作用的近五年研究进展[J];食品研究与开发;2014年19期
2 徐平湘;陈芩f，
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