【作者】 沈丽波；

【导师】 钱会南；

【作者基本信息】 北京中医药大学 ， 中医基础理论， 2005， 硕士

【摘要】 目的:本研究以中医五脏藏神理论为依据,从理论和实验方面,对脾藏神理论进行了探讨。 方法:在理论方面,本研究从五脏藏神的角度出发,阐述了脾在五脏藏神理论中的重要地位,并通过对脾胃与神志相关理论的梳理,从脾胃纳化水谷,转枢气机、以及运化水湿方面归纳了脾胃与神志相关理论的机制。同时,本文还对脾藏意主思中,“意”和“思”的含义进行了探讨。 在实验方面,本研究以Wistar大鼠为研究对象,分为模型组、正常组、治疗组(造模加灌胃归脾汤)、A方对照组(造模加灌胃柴胡疏肝散)、B方对照组(造模加灌胃天王补心丹)。采用苦降泻下、饮食失节加劳倦过度的方法建立脾虚模型,通过水迷宫实验、跳台实验、旷野实验和自发活动实验观测脾虚大鼠的行为变化,并用免疫组化方法检测生长抑素、胆囊收缩素、P物质和血管活性肠肽在海马CA1区、前额叶皮层和下丘脑腹侧核的变化。所有数据用SPSS10.0软件进行统计分析。 结果 (1) 造模结果:模型组大鼠从造模第3天起,逐渐出现倦怠懒动、拱背、扎堆、眯眼、或见颤抖等情况,皮毛疏松粗糙无光泽,且颜色发黄、易脱落,足蹼、鼻尖、耳缘粘膜色淡,身体消瘦,肛门周围充血、湿润,大便稀溏,或时干时溏,拉尾排便试验呈阳性。治疗组在第一周也出现类似的情况,但此后症状逐渐减少消失,体重增长迅速。模型组的体重增长缓慢,体重较轻,与正常组比较,差异极显著(P<0.01),说明造模较成功;而治疗组体重增长速度较快,体重较重,与模型组比较,差异极显著(P<0.01),说明治疗组所用的归脾汤对脾虚症状有明显的改善作用。而用于对照组的两种方药则无此疗效。 (2) 行为学观测结果:在水迷宫实验中,模型组大鼠游全程的时间较长,错误次数较多,与正常组比较,差异极显著(P<0.01),说明脾虚大鼠学习记忆能力下降。而治疗组大鼠游全程的时间较短,错误次数较少,与模型组比较,差异显著(P<0.01),说明归脾汤可以改善脾虚大鼠学习记忆能力。在跳台实验中,模型组大鼠错误次数较多,触电潜伏期短,与正常组比较,差异极显著(P<0.01),说明脾虚大鼠学习记忆能力下降。而治疗组大鼠错误次数较少,触电潜伏期长,与模型组比较,差异极显著(P<0.01),说明归脾汤可以改善脾虚大鼠的学习记忆能力。在旷野实验中,模型组理毛次数少,站立次数少,站立时间短,行走格数少,与正常组比较,差异极显著(P<0.01),说明脾虚大鼠的中枢兴奋性降低;而治疗组大鼠模型组理毛次数多,站立次数多,站立时间长,行走格数多,与模型组比较,差异极显著(P<0.01),说脾藏神理论探讨—脾虚模型行为与脑内神经肤变化相关机制研究明归脾汤可以改善中枢神经的抑制状态。在自发活动实验中,模型组理毛次数少,站立次数少,站立时间短,行走格数少,行走时间短,与正常组比较,差异极显著(P<0.01),说明脾虚大鼠的中枢兴奋性有所降低;而治疗组大鼠模型组理毛次数多,站立次数多,站立时间长,行走格数多,行走时间长,与模型组比较,差异极显著(P<0.01),说明归脾汤可以调节中枢神经的抑制状态。 (3)生长抑素、胆囊收缩素、P物质和血管活性肠肤的免疫组化研究结果:胆囊收缩素:在光镜下,CCK免疫阳性反应物呈棕黄色,免疫阳性细胞呈圆形或三角形,在海马、皮层、下丘脑均有分布。正常组显色较深,呈强阳性,界限明显;模型组显色淡,界限不明显,在海马、皮层及下丘脑免疫阳性反应物数量较少。光密度测量统计结果表明,CCK免疫阳性反应物在海马CAI区、前额叶皮层和下丘脑腹侧核平均和积分光密度均较低,与正常组比较,差异极显著(P<0. 01),说明脾虚大鼠学习记忆能力下降的作用机理与CCK改变有关;而治疗组显色较深,界限明显,在海马数量较多,且排列整齐,皮层及下丘脑免疫阳性反应物数量较多,光密度测量统计结果表明,其海马CAI区、前额叶皮层和下丘脑腹侧核光密度较高,与模型组比较,差异极显著 (P<0.01),说明归脾汤在改善脾虚大鼠学习记忆能力的作用机理与其对CCK的调节作用有关;A方对照组与B方对照组在光镜下形态变化与模型组相似,其统计数据与模型组无显著差异。 生长抑素:在光镜下,SOM免疫阳性反应物呈棕黄色,免疫阳性细胞呈圆形,在海马、皮层、下丘脑均有分布。正常组显色较深,呈强阳性,界限明显;模型组显色淡,界限不清,在海马数量少,且排列紊乱,皮层及下丘脑免疫阳性反应物数量较少,光密度测量统计结果表明,其在海马CAI区、前额叶皮层和下丘脑腹侧核光密度较低,与正常组比较,差异极显著(P<0.01),说明脾虚大鼠学习记忆能力下降的作用机理与SOM的改变有关;而治疗组显色均匀,界限明显,在海马数量较多,且排列整齐,皮层及下丘脑免疫阳性反应物数量较多,光密度测量统计结果表明,其海马CAI区、前额叶皮层和下丘脑腹侧核光密度较高,与模型组比较,差异极显著(P<0.01),说明归脾汤在改善脾虚大鼠学习记忆能力的作用机理与其对SOM的调节影响有关;B方对照组SOM在海马CAI区和下丘脑腹侧核光密度较高,与模型组比较,差异极显著(P<0.01),而与正常组比较,无显著差异。推测天王补心丹改善学习记忆的功能可能与归脾汤有相似的机理;A方对照组在光镜下形态变化与模型组相似,统计数据与模型组无显著差异。 P物质:在光镜下,SP免疫?更多还原

【Abstract】 Aim: The paper is to study the relationship of Spleen Zang and the Spirit, which is based on the theory of Five Zang-Spirit, by observing the behavior and detecting the neuropeptide of the rat model of spleen deficiency syndrome.Methods: To study the theory, the paper set forth the importance of the Spleen Zang in the spirit, and it’s mechanism. It discuss the deeper meaning of the "Yi"spirit and "Si"spirit which includes the meaning of learning and memory, and the relationship of the spleen and the learning and memory.The recognition considers that the Spleen Zang is the basis of the the acquired root and is the source of qi and blood production which transformation is the material foundation of spirit. The spleen governs the pivot of qi dynamism and the water metabolizing. The function of the spleen is the basis of the learning and memory.The experimental research proceeds from the angle of the influence of spleen deficiency on cognition. The spleen-asthenia rat model are produced by applying multiple factors such as abnormal diet, over strain and damage from abuse diarrhea drugs. Guipi decoction is used as a remedy to verify the availability of the model. Chaihushugan decoction and Tianwangbuxin decoction is used as a positive contrast. Water maze ,step-down method open field method and spontaneity method are used to study the behavior of the rats. Immunohistochemical method is employed to detect the variance of appearance and quantity of somatostatin(SOM), substance P(SP), cholecystokinin(CCK) and vasoactive intestinal peptide(VIP). All of these are done to reveal the effect of spleen deficiency on the mechanism of the learning and memory of a rat and to embody the significance of "the spleen storing the mind and governing thought". Result:(1) The result shows that the spleen deficiency group is obviously weaker than the normal group, the treated group is much better than the spleen deficiency group. The obvious difference is existed between the nomal group and the spleen deficiency group, and also between the treated group and the spleen deficiency group, Which clues to the weakness of the spleen deficiency rat and the curative effect of the Guipi decoction.(2)Through the water maze, step-down, open-field and spontaneity method, the decline in the learning and memory of the spleen deficiency rat is shown. The obvious difference is existed between the nomal group and the spleen deficiency group, and also between the treated group and the spleen deficiency group, Which clues to the decline of the spleen deficiency rat in learning and memory, and the curative effect of the Guipi decoction.(3) Immunohistochemical experiment showed that the quantity of CCK, SOM, SP and VIP have decreased obviously in the spleen deficiency group. However, the treated group did not show manifestation of decreasing in the quantity of CCK, SOM, SP and VIP neuropeptide, and compared with spleen deficiency group the difference is obvious which clues to the mechanism of the decline of the learning and memory.Conclusion:(1) The Spleen Zang-Spirit theory is an important part of the Wu Zang-Spirit theory. The Spleen Zang is the basis of the the acquired root and is the source of qi and blood production which transformation is the material foundation of spirit, and governs the pivot of qi dynamism and the water metabolizing.(2) The function of the spleen is the basis of the learning and memory.(3) The primary method to make the spleen deficiency model is used, which proved to be success.(4) Through the water maze, step-down, open-field and spontaneity method, the decline in the learning and memory of the spleen deficiency rat is shown, Which clues to the decline of the spleen deficiency rat in learning and memory, and the curative effect of the Guipi decoction.(5) The decreasing in the quantity of CCK, SOM, SP and VIP neuropeptide of the spleen deficiency rat’s brain, clues to the mechanism of the decline of the learning and memory.更多还原