头穴丛刺对慢性酒精中毒大鼠学习记忆能力及海马HIF表达的影响

发布时间：2018-01-13 21:12

  本文关键词：头穴丛刺对慢性酒精中毒大鼠学习记忆能力及海马HIF表达的影响　出处：《黑龙江中医药大学》2013年博士论文　论文类型：学位论文

  更多相关文章： 慢性酒精中毒 学习记忆障碍 海马 头穴丛刺 盐酸多奈哌齐 VEGF HIF-1α

【摘要】：目的：通过观察头穴丛刺法对慢性酒精中毒所致学习记忆障碍模型大鼠学习记忆能力、HIF-1α及其靶基因VEGF mRNA和HIF mRNA表达的影响,探讨头穴丛刺对慢性酒精中毒所致学习记忆障碍的治疗效果及其作用机制,为慢性酒精中毒所致脑病的临床治疗提供一定的实验基础和理论依据。 方法：将成年雄性Wistar大鼠100只,随机分为空白对照组和酒精灌胃组。酒精灌胃组采用酒精灌胃法制备慢性酒精中毒所致学习记忆障碍大鼠模型,灌酒6周,通过Morris水迷宫检测大鼠的学习记忆能力。再将慢性酒精中毒所致学习记忆障碍造模成功的大鼠随机分为模型组、多奈哌齐组和头穴丛刺组。多奈哌齐组给予多奈哌齐0.05mg/kg灌胃,1次/日,治疗4周。空白对照组以同等体积的生理盐水灌胃,灌胃量为10ml/kg,1次/日,共灌胃4周。头穴丛刺组采用头穴丛刺法,取百会和百会左右旁开2mm三个穴位,留针30min,1次/日,治疗4周。治疗4周后,采用Morris水迷宫检测大鼠的学习记忆能力；光镜下观察海马组织病理学变化；免疫组化检测海马HIF-1α、VEGF阳性表达；采用实时荧光定量-PCR(real-time fluorogentic quantitative PCR, RFQ-PCR)法检测海马HIF-1αmRNA、VEGFmRNA表达。 结果： 1.Morris水迷宫检测结果：造模6周后,灌酒模型组大鼠学习记忆能力明显下降,与空白对照组相比差异显著(P0.05)；治疗4周后,多奈哌齐组和头穴丛刺组大鼠的学习记忆能力与模型组相比有所提高,具有显著性差异(P0.01)；多奈哌齐组和头穴丛刺组相比无显著性差异(P0.05)。 2.HE染色结果：空白对照组未见明显病理学变化,模型组可见大量神经元细胞固缩,胞体形态不规则,胞浆浓缩深染,核仁变小或消失,细胞尥周围可见明显的空壳区；多奈哌齐组和头穴丛剌组神经细胞排列层次及细胞形态较清晰,少见固缩细胞,与模型组相比腑组织损害程度明显减轻；而多奈哌齐组与头穴丛刺组相比,固缩细胞数量明显多于头穴丛刺组。 3.HIF-1α、VEGF免疫组化染色结果：正常对照组神经细胞数量、形态分布正常,排列紧密有序,细胞核圆而大,染色浅,核仁清晰,可见少量HIF-1α、VEGF阳性细胞表达；模型组、多奈哌齐组和头穴丛刺组均可见HIF-1α、VEGF阳性细胞增加,与正常对照组比较差异具有统计学意义(P0.05,P0.01)；而多奈哌齐组和头穴丛刺组较模型组阳性细胞表达减少,差异具有统计学意义(P0.05,P0.01)；多奈哌齐组与头穴丛刺组相比,后者减少更为明显,差异具有统计学意义(P0.01)。 4.HIFmRNA、VEGF mRNA real-time RT-PCR检测检测结果：空白对照组HIFmRNA、VEGF mRNA表达极弱；在模型组、多奈哌齐组和头穴丛刺组中,HIFmRNA、VEGF mRNA均呈显著强表达,高于空白对照组；而多奈哌齐组和头穴丛刺组的HIFmRNA、VEGF mRNA表达较模型组弱；头穴丛刺组与多奈哌齐组相比,后者表达更高。 结论： 1.采用酒精灌胃法制作慢性酒精中毒所致学习记忆障碍大鼠模型,并利用Morris水迷宫从行为学和HE染色从组织学两方面对其进行评价,模型成功、可靠。 2.头穴丛刺能够改善慢性酒精中毒所致学习记忆障碍大鼠的学习记忆障碍,改善海马组织病理学变化,能够抑制神经元凋亡,减少神经细胞病理变化,从而改善学习记忆能力。 3.头穴丛刺能够降低HIF-1α和VEGF的阳性细胞表达,减少脑损伤后的脑组织神经元损伤,减少神经元凋亡,发挥脑保护功能。 4.头穴丛刺可应用于慢性酒精中毒所致学习记忆障碍的防治,有望成为慢性酒精中毒所致学习记忆障碍新的治疗方法之一。
[Abstract]:Objective: to chronic alcoholism induced learning and memory impairment of learning and memory ability of rats by observing the CASP, affecting the expression of alpha and its target gene VEGF mRNA and HIF mRNA HIF-1, to investigate the scalp acupuncture on chronic alcoholism induced learning and memory impairment in the therapeutic effect and its mechanism, provide theoretical and experimental basis on the basis of some clinical treatment for chronic alcoholism induced encephalopathy.
Methods: 100 male Wistar rats, were randomly divided into control group and alcohol gavage group. Intragastric group with alcohol by chronic alcoholism induced learning and memory impairment in rats model of gastric lavage method, alcohol for 6 weeks, the learning and memory of rats in Morris water maze test can then. Chronic alcoholism induced impairment of learning and memory of rats were randomly divided into model group, donepezil group and scalp acupuncture group. Donepezil group were given donepezil 0.05mg/kg orally, 1 times / day, for 4 weeks. Blank control group with the same volume of saline by gavage, gastric lavage volume is 10ml/kg, 1 time / day, a total of 4 weeks. Head acupuncture group with head acupuncture method, Baihui and Baihui about 2mm beside the three points for 30min, 1 times / day, for 4 weeks. After 4 weeks of treatment, the ability of learning and memory in Morris water maze test in rats were observed; in the light microscope Histopathological changes, immunohistochemistry, HIF-1 positive expression of VEGF and -PCR in hippocampus were detected, and HIF-1 mRNA mRNA and VEGFmRNA expression in hippocampus were detected by real-time fluorogentic quantitative PCR (RFQ-PCR).
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