【作者】 张筱军；

【导师】 周永学；

【作者基本信息】 陕西中医学院 ， 方剂学， 2008， 硕士

【摘要】 目的:在既往药效学实验和临床研究证明硝菔通结颗粒能有效治疗便秘的基础上,探讨便秘的中医病理机制及硝菔通结颗粒的立法原则与处方配伍特点。同时,通过观察便秘小鼠排黑粪时间及数量情况,以反映该药物泻下作用的强弱;通过观察黑色碳末在便秘小鼠小肠及大鼠大肠的推进距离,以反映该药对肠道平滑肌的影响,从而判断其对大鼠肠道运动的作用;通过测量便秘小鼠不同肠段水分的含量,以反映该药对大鼠肠道的水分含量的增加情况,从而判断其主要作用部位;通过观察硝菔通结颗粒对便秘大鼠血浆MTL、SS含量的变化,以从放免角度阐明该药对便秘的作用机制。方法:理论探讨:搜集并总结历代论述便秘的相关文献,同时结合现代医学对便秘的研究进展,探讨便秘的中医病因、病机、理法方药及硝菔通结颗粒的立法原则与配伍特点。实验研究:1.对便秘小鼠排便时间及粒数的影响:将小鼠60只随机分为对照组、模型组、麻仁丸组、硝菔通结颗粒大、中、小剂量组,每组10只。除对照组外,其余各组灌服地芬诺酯混悬液复制便秘模型成功后,对照组和模型组每天给予生理盐水灌胃,其余各组灌食含有2%墨汁的相应液体。给药后连续观察8h,记录小鼠第一次排黑便时间,4h内、8h小鼠排出干粪便总粒数、干粪便重量、稀粪点数和不排便动物数。2.对便秘小鼠小肠运动的影响:分组及给药同实验一。末次给药30min后,各组小鼠均按0.02ml/g的剂量给予碳末混悬液,再过后20min,小鼠脱颈椎处死,测量幽门至回肠部的长度及从幽门至墨汁前沿的距离,计算墨汁推进百分率,并注意观察各组小肠容积是否增大。3.对便秘大鼠大肠动力的影响:将60只大鼠禁食不禁水12h后麻醉,在腹中线做一切口,把塑料管由回盲部插入结肠1cm,结扎固定,并通过肌肉皮肤从背部引出,以备给药。次日将60只大鼠分为6组,分组、造模方法、各组所给药物均同实验一。末次给药30min后,所有大鼠灌注含有2%墨汁的药液后立即计时;另外60只所灌食液中不含色素指示剂。灌胃后立即浅麻醉,沿腹中线做切口找到回盲部,灌注含墨汁的生理盐水。在靠近回盲部的回肠用双线做结扎,在两线之间剪断回肠并计时。10分钟后将大鼠处死,测量结肠总长度和墨汁推进距离,计算墨汁推进百分率。4.对便秘小鼠不同肠段水分吸收的影响:动物、分组及给药量均同实验一。给药后2h将小鼠处死,后剪开肠系膜,自幽门下端用线结扎,剪断肠管;再于回盲部上边结扎,剪断肠管,取出该段小肠。再于直肠末端结扎,剪断肠管,取出大肠。分别称量各鼠的小肠和大肠湿重并记录,然后将各鼠小肠和大肠放在干燥箱里烘干,取出称重。用公式计算肠内水分含量。5.对便秘大鼠血浆及肠组织中MTL、SS含量的影响:动物、分组及给药量均同实验三。各组血浆标本的留取:用药结束后,各组大鼠麻醉,股静脉采血2ml,注入特殊料管中混匀,离心15min,取血浆250μl,-20℃以下保存待检。各组肠组织匀浆标本留取:上述大鼠股静脉采血后,迅速剖腹,在十二指肠与空肠交接取0.5cm长的肠组织,立即液氮冻存。测胃动素样品处理:小肠组织加入冰醋酸1ml,煮沸10min,匀浆,加入PBS1ml,同上方法保存。测生长抑素样品处理:小肠组织于煮沸的9g/L氯化钠溶液1ml中煮沸3min,冷却后加入冰醋酸0.5ml于匀浆器中制成匀浆,再用氢氧化钠溶液0.5ml中和,同上方法保存。采用放射免疫分析非平衡法测定。结果:理论研究:便秘的主要病位在肺、脾、胃、大肠,气阴亏虚,推动乏力,肠道失润,糟粕停聚为慢性便秘的主要病机,治当益气养阴,软坚润燥,运下通便。实验研究:1.对便秘小鼠排便时间及粒数的影响:与模型组比较,对照组、XFTJH、XFTJM、XFTJL及麻仁丸组均能缩短便秘小鼠首次排便的时间、增加排便重量和排便粒数(P<0.05)。与麻仁丸组比较,XFTJH有显著性差异(P<0.05)。2.对便秘小鼠小肠运动的影响:与模型组比较,XFTJH、XFTJM对便秘模型小鼠的小肠蠕动和推进速度有非常显著性差异(P<0.01),呈剂量依赖性;与麻仁丸组比较,则差异亦有显著性(P<0.05)。3.对便秘大鼠大肠动力的影响:与模型组比较,XFTJH、XFTJM对便秘模型大鼠的大肠蠕动和推进速度有非常显著性差异(P<0.01);与麻仁丸组比较,XFTJH、XFTJM有显著性差异(P<0.05)。4.对便秘小鼠不同肠段水分吸收的影响:与模型组比较,XFTJH、XFTJM、XFTJL、麻仁丸组均可使小鼠大肠水份含量增加(P<0.05),而以XFTJH最为明显(P<0.01);XFTJH亦可使小鼠小肠水份含量增加,与模型组比较有统计学意义(P<0.05),而麻仁丸组对其小肠水份含量没有影响(P>0.05);与麻仁丸组比较,XFTJH、XFTJM对鼠大肠水分含量有差异性(P<0.05)。5.对便秘大鼠血浆及肠组织中MTL、SS含量的影响:在灌喂XFTJ后,血浆及肠组织中MTL明显增加,SS明显降低,与模型组比较有统计学意义(P<0.05),尤以大剂量最为显著(P<0.01),与麻仁丸组比较,只有硝菔通结大剂量有显著性(P<0.05)。结论:XFTJ组方严谨,针对病机,补通结合,升降同用,标本兼顾,益气健脾而不滋腻,运肠通便而不峻攻,清热润燥而不寒凉,为治疗便秘的有效新制剂。该药可减少便秘小鼠排便时间,增加排便粒数;可提高便秘小鼠小肠运动,增加炭墨推进率;既能直接作用于便秘大鼠的大肠,亦可在小肠吸收后作用于大肠,增加其大肠推进作用;可增加便秘小鼠大肠和小肠内水分含量;能增加便秘大鼠血浆及肠组织中MTL、减少SS含量,这可能是硝菔通结颗粒提高肠动力,改善便秘症状的机制。更多还原

【Abstract】 Objective: In the past pharmacodynamics of experiments and clinical studies have proved that Xiaofutongjie Grain can effectively on the basis of the treatment of constipation,Constipation of the pathological mechanism of Chinese medicine and Xiaofutongjie Grain of the legislative principle of prescription and compatibility characteristics.At the same time,Pai constipation mice by observing the time and the number of black fecal situation,To reflect the role of the drug under the strength of diarrhoea;By observing carbon black at the end of the small intestine constipation mice and rats from the advance of colon,On the drug to reflect the impact of intestinal smooth muscle,and thus judged on the role of rat intestinal movement.Constipation mice by measuring the water content of different Changduan to reflect the drug on rat intestinal increase in the water content,to determine the role of the main parts.Xiaofutongjie Grain observation of constipation guitar particles in plasma MTL,SS content changes,immune from the group to clarify the perspective of the drug on the mechanism of constipation.Methods:Academic exploring:Collect and sum up the history of literature related to constipation-related expositions,and in accordance with years of clinical trials,combined with modern medical research on the progress of constipation,Constipation of the Chinese etiology,pathogenesis,and Lifa Prescription Xiaofutongjie Grain of the legislative principle of particle characteristics and Compatibility.Experimental Study:1.Constipation mice defecation on time and the impact of grains: 60 mice were randomly divided into control group,model group,Maziren pill group,XFTJH, XFTJM,XFTJL,each group of 10.In addition to the control group,the rest of the group administered DC suspension replication model after the success of constipation,Model group and the control group given saline every day gavage,the group fed with the remaining 2% of the corresponding liquid ink.After administration,continuous observation of 8h,recorded the first row and mice have time,4h,8h mice stem from the faeces of grains, dry stool weight,defecation Dilute stool points and the number of animals. 2.Constipation mice on the movement of the small intestine:With the experimental group and an administration.At the end of 30min after administration,according to the mice 0.02ml/g at the end of the dose given carbon suspensionTo 20min after the mice were sacrificed from cervical measurement pylorus to the length of the ileum and ink from the pylorus to the forefront of distance,and the percentage of ink and attention to the small intestine in each group was observed whether the increased volume.3.Constipation rat colon driving force of the impact:Will be 60 rats do not give food but drinking water can,and 12h after anesthesia,In the womb line with a scalpel incision to do the plastic pipe from the ileocecal inserted colon,a depth of 1cm,Fixed ligation,Through the skin from the back muscles leads to prepare for administration.The next day will be 60 rats were divided into 6 groups,sub-modeling methods,the groups of drugs given in the first experiment with the same.The last time 30 min after the end of treatment,All the rats fed food containing 2% of ink to End liquid immediately after time,Another 60 by the feeding of non-liquid pigment indicator.Gavage immediately after light anesthesia,VSA incision along the lines do find ileocecal,Infusion of normal saline containing ink. In and around the ileum ileocecal ligation done with two-lane,cut in two between the lines and ileum time.Rats were sacrificed after 10 minutes,the total length of the colon and measurement ink promote distance,and the percentage of ink.4.Changduan constipation mice on the different absorption of water:Animals,both with the administration and the division of one.2h after administration to mice were sacrificed after mesenteric cut,with the bottom line since ligation Helicobacter pylori, intestinal cut;Further upside in the ileocecal ligation,cut intestine,and the removal of the small intestine.Ligation of the rectum to the end,cut intestine,large intestine removed. Weighing all the rats were small intestine and colon wet weight and recorded,and then the rat small intestine and colon,on the oven drying, and weighing out.5.Constipation in the plasma and intestinal tissue in MTL,the content of SS:Animals, both with the administration and the group of three.Plasma samples of each group collecting:After the treatment, the rats anesthesia, 2 ml femoral vein blood injected into the cylinder in the special blending, centrifugal 15 min,the plasma from 250μl,to be kept below -20℃seized.Each group intestinal tissue specimens Collecting:The rat femoral vein blood,and rapidly belly,in the duodenum and jejunum transfer of 0.5cm in length from the intestinal tissue and immediately frozen in liquid nitrogen.Measurement motilin sample processing:small intestine organizations to join the 1ml of acetic acid,and boil for 10min, homogenized,by adding PBS 1ml,ibid. Preservation methods.Somatostatin test sample processing:small intestine in boiled 9g/L NaCl in 1ml boiled for 3 min,cooling after acetic acid by adding 0.5ml of homogenate was made in the homogenate,and then 0.5ml sodium hydroxide solution and in,ibid.preservation methods.By RIA analysis of non-equilibrium method.Results:Academic exploring:Constipation disease in large intestine,spleen, stomach,lung incidence of disease is the major factor Qilin deficiency,and promote fatigue,loss of intestinal Run,stop the dregs for the constipation-pathological mechanism of the main Chinese medicine,treatment should be Yiqiyangyin,soft Kennedy Runzao,Wan under purge.Experimental Study:1.Constipation mice defecation on time and the impact of grains: Compared with model group,the control group,XFTJH, XFTJM, XFTJL and Maren Pills group could shorten constipation mice defecation first time,increase the weight and defecation defecation grains(P<0.05). And Maren Pills group,XFTJH There was a significant difference(P<0.05).2.Constipation mice on the movement of the small intestine:Compared with model group,XFTJH,constipation mice XFTJM small peristalsis and promote speed there was a significant difference(P<0.01) in a dose-dependent manner compared with Maren Pills, there has been a significant difference(P<0.05).3.Constipation rat colon driving force of the impact: Compared with model group, XFTJH,constipation XFTJM rat model of the large bowel movements and speed of advance there was a significant difference(P<0.01) compared with Maren Pills,XFTJH, XFTJM There was a significant difference(P<0.05).4.Changduan constipation mice on the different absorption of water:Compared with model group,XFTJH,XFTJM,XFTJL,Maren Pills group can make mice colorectal water content increased(P<0.05),and to XFTJH most significantly(P<0.01);XFTJH mouse small intestine so that water can be increase in the content,compared with the model group with statistical significance(P<0.05), while Maren Pills group to the small intestine does not affect the water content(P>0.05),compared with Maren Pills,XFTJH,the rat colon XFTJM Moisture content differences(P <0.05).5.Constipation in the plasma and intestinal tissue in MTL,the content of SS:Hey XFTJ in irrigation,plasma and intestinal tissue significantly increased in MTL,SS decreased significantly compared with the model group with statistical significance(P<0.05), especially in large doses the most significant(P<0.01),and Maren Pills Group,the only XFTJH was significant(P<0.05).Conclusion:XFTJ set of stringent,in view of machines,fill-combined with the same take-off and landing,were balanced,Yiqi Jianpi without mayonnaise greasy,Wan Jun attack intestinal catharsis without heat Runzao not Hanliang, effective for the treatment of constipation new agents.The drug can reduce constipation mice defecation time and increased defecation grains;constipation mice can increase intestinal movement,promoting the rate of increase in carbon Mexico;constipation can direct role in the rat large intestine, can be absorbed in the small intestine role in the large intestine,increase its role in promoting colorectal;increase constipation large intestine and small intestine of mice moisture content;constipation increase in plasma and intestinal tissue in MTL to reduce SS content,this is probably XFTJ increase intestinal motility and improve the symptoms of constipation mechanism.更多还原