【作者】 刘晓霓；

【导师】 牛欣； 司银楚；

【作者基本信息】 北京中医药大学 ， 中西医结合基础， 2004， 博士

【摘要】 本课题为国家中西医结合基础重点学科211工程资助项目(NO 2001-004)，本研究主要探讨半夏泻心汤、生姜泻心汤和甘草泻心汤对胃运动影响的作用机理和部分物质基础。 理论研究 通过对类方研究、胃运动研究以及中药复方物质基础近年来的研究概况进行回顾性总结，我们提出了中药复方类方研究策略。中药复方类方研究策略的内容包括：中药复方类方差异点的研究和中药复方类方内在固有规律的研究。中药复方类方研究策略的目的：为中药复方的配伍规律和物质基础提供实验依据；为重组高效中药复方提供新思路。本研究以半夏泻心汤、生姜泻心汤和甘草泻心汤为研究对象，以胃运动为研究切入点，对三方胃运动影响的作用机理和物质基础进行较为深入的探讨，以期为中药复方类方研究提供有意义的科研思路和实验依据。 实验研究分三部分 第一部分 目的：本部分实验主要观察半夏泻心汤、生姜泻心汤、甘草泻心汤对在体大鼠胃排空作用的影响并对其作用机理进行了深入探讨。 结果：1)对胃排空的影响：半夏泻心汤对胃排空有促进作用；生姜泻心汤对胃排空为抑制作用；甘草泻心汤对胃排空有促进趋势。2)对胃肠激素的影响：半夏泻心汤可以使大鼠血中VIP水平下降、P物质水平下降、SS水平升高。生姜泻心汤可以使MTL水平升高，GAS水平升高，SS水平升高。甘草泻心汤可以使MTL水平升高，GAS水平升高。对三个泻心汤组大鼠胃内色素相对残留率和血中激素水平进行相关分析，结果发现半夏泻心汤促进胃排空可能与其降低大鼠血中VIP水平有关；生姜泻心汤对胃排空的抑制作用可能和其升高大鼠血中VIP和SS的水平有关，甘草泻心汤促进胃排空的趋势可能与大鼠血中MTL水平升高有关。3)对平滑肌收缩调控抑制蛋白及其基因的影响：Calponin和caldesmon为平滑肌收缩的两个调控抑制蛋白。本实验显示calponin和caldesmon对胃肠道平滑肌有不同的调节作用，可能介导了三泻心汤对胃运动的调节作用。三泻心汤组大鼠胃窦平滑肌calponin和caldesmon表达量均有下调趋势。免疫组化结果显示：calponin表达量为生姜泻心汤组>甘草泻心汤组>半夏泻心汤组。与对照组相比，半夏泻心汤组calponin表达下调明显。caldemon表达量为甘草泻心汤组>生姜泻心汤组>半夏泻心汤组。与对照组相比，三泻心汤组caldemon表达量下调均较为显著。RT-PCR结果显示：与对照组相比，三泻心汤组calponin mRNA和caldesmon mRNA表达量也有所下调，下调结2半夏泻心汤及其类方对胃运动影响的作用机理及物质基础研究果与免疫组化结果大致相同。 第二部分 目的:本部分实验观察三方含药血清对胃窦平滑肌细胞收缩功能的影响，在细胞水平深入探讨三方对胃运动影响的作用机理。 结果:1)对胃窦平滑肌细胞收缩功能的影响:三泻心汤含药血清可以引起大鼠胃窦平滑肌细胞不同程度的收缩，结果显示，三泻心汤含药血清以5%终浓度，作用时间5分钟为佳。从促进收缩的程度来讲，半夏泻心汤组>生姜泻心汤组>甘草泻心汤组。2)对胃窦平滑肌细胞内钙离子的影响:三泻心汤含药血清对胃窦平滑肌细胞内钙离子的荧光峰值影响的结果显示，与对照组相比，半夏泻心汤含药血清使胃窦平滑肌细胞内钙离子的荧光峰值明显增加，其他两方含药血清对细胞内钙离子的荧光峰值影响不明显。三方组间比较，细胞内钙离子荧光峰值大小顺序为半夏泻心汤组>生姜泻心汤组>甘草泻心汤组。连续扫描结果显示，在不同时间段，半夏泻心汤和生姜泻心汤含药血清均可以使大鼠胃窦平滑肌细胞内的荧光值增加，提示二泻心汤可以增加平滑肌细胞内的钙离子浓度。而组间比较显示，甘草泻心汤含药血清对胃窦平滑肌细胞内的荧光值改变不明显，但组内比较，有升高的趋势。”对胃宾平滑肌细胞收缩调控抑制蛋白墓因的影响:三泻心汤含药血清也均可以使胃窦平滑肌细胞。alPonin mRNA和ca ldesmon mRNA的表达下调，结果与整体实验相似，提示三泻心汤含药血清对胃窦平滑肌细胞的收缩作用，与其直接改变ca IPonin mRNA和caldesmonmRNA的表达量有关。 第三部分 目的:观察给药后一定时间大鼠血中三泻心汤小聚碱、黄答贰和甘草次酸的含量变化及三种成分与胃运动的关系;观察三泻心汤中三种有效成分小聚碱、黄答式和甘草次酸不同配比组成的有效成分组方对胃窦平滑肌细胞收缩功能的影响，在细胞水平探讨三泻心汤部分有效成分配伍规律。 结果:1)大鼠血中三种成分含t:小聚碱含量大小依次为生姜泻心汤组>半夏泻心汤组>甘草泻心汤组，但差别无统计学意义;黄答戒含量大小依次为生姜泻心汤组>半夏泻心汤组>甘草泻心汤组，且生姜泻心汤组大鼠血中黄琴贰含量远远高于半夏泻心汤组和甘草泻心汤组;甘草次酸含量依次为生姜泻心汤组>甘草泻心汤组>半夏泻心汤组，生姜泻心汤组大鼠血中甘草次酸含量远远高于半夏泻心汤组。对三泻心汤组大鼠胃内色素残留率和大鼠血中三种成分的含量之间相关分析结果表明，半夏泻心汤组的促进大鼠胃排空作用可能与大鼠血中小栗碱含量有关，而生姜泻心汤组抑制胃排空的作用可能与大鼠血中?更多还原

【Abstract】 The fund of this study (No 2001-004) is supported by the 211 project of national key subject of combination of traditional Chinese and western medicine, mainly probing the mechanism and material basis of banxia(BX) , shengjiang(SJ), gancaoxinxin(GC) decoctions for the gastric motility .Theoretical study:A review of study on analogous preparations ,material basis of compound prescription and gastric motility was summarized to provide a tactics of study on compound prescription and its analogous. In this study we chose BX, SJ, GC decoctions as the object to explore their material basis and mechanism for the gastric motility.Experimental study:PartiObjective: To observe the capacity of gastric evacuation of rats by given BX, SJ, GC decoctions and explore the mechanism of these decoction for gastric motility.Results: A promotive effect of BX decoction on the gastric evacuation was observed; a inhibitive effect of SJ decoction on the gastric evacuation was showed .a trend of promotion in GC decoction for gastric evacuation appeared.These decoctions have different influence on the gastrointestinal hormones: there was a marked drop of the levels of VIP,SP and a rise of SS in rat serum in group BX decoction. The SJ decoction could raise the levels of MTL,GAS and SS in rat serum. The levels of MTL,GAS in rat serum were also increased in group GC decoction. Correlation analysis between Gastric residue of dextran blue-2000 and gastric evacuation showed: There was a negative correlation in group BX decoction between gastric residue of dextran blue and level of VIP in rat serum; There was a positive correlation in group SJ decoction between Gastric residue of dextran blue and levels of VIP and SS in rat serum; there was a positive correlation between Gastric residue of dextran blue and levels of MTL in rat serum.Calponin and caldesmon were both the important proteins regulating thecontraction of smooth muscles. This study showed these two proteins could regulate the status of smooth muscles of stomach. Maybe the regulating effect of BX, S J, GC decoctions on gastric motility was partly induced by calponin and caldesmon .the express of calponin and caldesmon all had decreased trend in group BX,SJ,GC decoctions vs control group . The results of histochemistry showed: the express content of calponin in group S J decoction was the first,group GC decoction was the second and group BX decoction was the last, the express content of calponin in group BX decoction decreased greatly vs group control, the express content of caldesmon in group GC decoction was the first ,group SJ decoction was the second and group BX decoction was the last, the express content of caldesmon in three groups were all decreased obviously vs group control. The results of PT-PCR was similar to the ones of histochemistry.Part 2Objective: To study the contractive effect of three decoctions-contained serum on antral smooth muscle cells .Results: Three decoctions-contained serum could arised different contraction of rat antral smooth muscle cells. Good contractive effect showed when given 5% final concentration of serum and reacted for 5 min. the degree of contraction in group BX decoction was the first, group SJ decoction was the second and group GC decoction was the last.Different peak fluorescent value of intracellular calcium in rat antral smooth muscle cells was observed in three decoctions-contained serum groups. The peak value in group BX decoction was the first, group SJ decoction was the second and group GC decoction was the last. Peak fluorescent value increased greatly in group BX decoction vs group control. The results of scanning continuously were as follows: three decoctions-contained serum could increase the fluorescent value of cells obviously.Three decoctions-contained serum could decrease the express of calponin mRNA and caldesmon mRNA in rat antral smooth muscle cells.Part3Objective:To observe the three effective constituents content of three decoctions in rat plasm and analysis the relationship between the effective更多还原