【作者】 马飞翔；

【导师】 甘君学；

【作者基本信息】 南京中医药大学 ， 针灸推拿学， 2009， 硕士

【摘要】 目的:对便秘大鼠粪便采用水份烘干、pH测定和定性加半定量细菌培养的方法检测泻剂结肠大鼠粪便水份、pH、细菌含量情况,组间相互对照,结合雌雄对照等对照,观察针刺对泻剂结肠大鼠粪便的影响,初步探讨针刺治疗慢传输型便秘(STC)的机制及针刺的有效性。方法:1、动物分组:选用造模成功的SD大鼠60只,雌雄各半。按照完全随机化方法和性别分为针刺组、药物组、针药结合组、假针刺组、模型组5大组10小组,分组后各组间大鼠体重、性别无差异。正常对照组的SD大鼠12只,雌雄各6只,按性别分为2组。2、模型建立:造模大鼠每日定时应用已打粉过80目筛的大黄细粉悬浊液进行灌胃。首次给药剂量为150mg/kg·d,以后每日按150mg/kg·d递增,至50%大鼠达到腹泻时,维持此剂量至80%大鼠稀便消失。续剂量再按每日150mg/kg·d递增至又见50%大鼠达到腹泻时再维持剂量再加量,如此循环三次灌胃饲养约2个月。待第三次80%的大鼠稀便消失,维持当时的剂量1周,之后停止给药并予普通软饲料,造模成功,停止造模。3、标本采集:分别收集停止灌胃期间、停止治疗后大鼠的粪便,进行水份分析、pH分析和细菌培养。4、统计处理:应用SPSS13.0统计软件,对数据进行统计分析。结果:1、应用大黄悬浊液按时定量灌胃可以对正常SD大鼠建立“泻剂结肠”模型,且该模型能较好的模拟虚秘(气阴两虚或阴阳两虚证)的若干指标;2、“泻剂结肠”大鼠①粪便水份含量显著降低,由正常的42%左右的含水量降至37%的含水率;②千克体重24小时粪便粒数亦显著降低,具体而言,由正常时63粒左右降至47粒左右;③粪便pH若偏碱性,由正常时的6.69升至6.87左右;④粪便存在菌群失调现象。粪便中腐生菌(大肠杆菌、大肠球菌)含量增加,相应的益生菌(双歧杆菌、乳酸杆菌)含量却较正常状态时降低。3、就升高粪便水份指标而言,针药结合的方法最佳,而针刺和药物的作用相当。提示临床上对于STC见于粪便干结或硬结的患者宜采取针药结合的方法;4、就提高千克体重24小时粪便粒数的作用而言,也是针药结合的作用最佳,针刺和药物的作用相当。提示临床上对于STC见于粪便量少或周期便次较少的患者宜采取针药结合的方法;5、便秘状态粪便pH略有增高,针灸和药物治疗的方法有效且疗效相当,其余方法对此无作用;6、STC患者停药自愈可能性小。不过,肠道大肠球菌有一定的自我修复能力。但是实验组中,只有单纯针刺才能有效降低大肠球菌含量;7、针刺和针药结合可以改善便秘状态大鼠大肠杆菌,且其效果无差异;8、实验组中只有针药结合才能提高便秘状态粪便中双歧杆菌的含量;9、实验组中只有单纯针刺才能提高便秘状态粪便中乳酸杆菌的含量;10、就性别而言,只有药物组在粪便水份含量和千克体重24小时粪便粒数指标上存在差异,且表现为雌性疗效优于雄性。究其原因可能为雄雌药物耐受程度不一样。提示我们在动物实验中对于雌性动物在给药剂量上应该适当降低雌性动物的剂量;11、综合给各观测指标,考虑临床医疗费用实际,我课题组建议:对于STC患者,首推治疗方法当为单纯针刺治疗;针药结合法适宜于患者便质干硬或量少时治标之用;当患者粪便菌群分析已经存在菌群失调的情况下,尽量避免口服西沙必利类似药物,可以应用含益生菌的制剂。更多还原

【Abstract】 Objective: We researched on the droppings of constipation rats with water drying, pH determination and qualitative add semi-quantitative bacterial culture methods to detect the moisture, pH, bacterial content of the cathartic colon rat droppings.Through inter-group cross-referencing and male and female control etc,we try to find out the effects of acupuncture on the cathartic colon rat droppings,and preliminary study on the mechanism and the effectiveness of acupuncture for the slow transit constipation (STC).Methods: 1.Grouping the animals: selecting 60 successful model SD rats, half male and female. Completely according with the random method of grouping and gender . Divided them into acupuncture group ,gender.drug group, with acupuncture and drug group, sham acupuncture group, model group.5 big groups 10 small groups.The split among the three groups of rats after the body weight, sex is no difference. SD of normal control group, 12 rats, 6 male and female each, divided into 2 groups by gender. 2.Set up the experimental model: model rats were Ig with the rhubarb powder suspension regular daily,the rhubarb powder has been through a 80 mesh sieve.The first dose is 150mg/kg·d,then progressively increase 150mg/kg·d, when 50% rats had diarrhea , maintain this dose to 80% rats’ diluted stool disappeared.And then continued progressively increase 150mg/kg·d, when 50% rats had diarrhea,maintain this dose to 80% rats’ diluted stool disappeared.Repeat the above steps again for two and a half months,when 80% rats’ diluted stool disappeared for the third time, maintain the dose for one week,then cessated drug delivery and offer these rats the ordinary soft-feed.The experiment model was successfully made,so we can stop setting up model. 3.Specimen collection: collecting rat droppings when stopped gavaging and treatment for moisture analysis, pH analysis and bacterial culture. 4.Statistical treatment: applicate SPSS13.0 statistical software to analysis the data.Results:1.Normal SD rats Ig with the rhubarb powder suspension regular daily can be used to establish "cathartic colon" models,and these models can simulate virtual Peru (2 Deficiency of both Qi and Yin deficiency or yin and yang) with a number of indicators;2."Cathartic colon" rats:①fecal water content significantly decreased, by about 42 percent of normal water content dropped to 37 percent moisture content;②24 hours stool grains per kg body weight also significantly reduced, specifically, by the normal of around 63 down to about 47;③fecal pH was alkaline,by the normal of 6.69 increase to 6.87;④stool is in a dysbacteriosis situation .Saprophytic fecal bacteria (Escherichia coli, Enterococcus large) content increased, the corresponding probiotic (Bifidobacterium, Lactobacillus) content is lower than the normal state;3.When it comes to increasing the fecal moisture , the best method is acupuncture combined with medicine, the effects of acupuncture and drugs are equal.This indicates that STC patients with stool colorectal or induration on clinical should adopt the method of acupuncture combined with drugs;4.When it comes to increasing the 24 hours stool grains per kg body weight, the best method is also acupuncture combined with medicine,and the effects of acupuncture and drugs are also equal.This indicates that STC patients with less stool volume or less stool frequency in certain time should adopt the method of acupuncture combined with medicine;5.Fecal pH is slightly higher than normal when constipation, acupuncture and drug treatment are both efficacy and the effects are equal,other methods have no effects on it;6.The possibility of STC patients’ self-healing when stop treatment is very small.But large intestinal enterococci have certain self-repair capacity.However,in these experimental groups, only simple acupuncture can effectively reduce the large Enterococcus content;7.Acupuncture and acupuncture combined with medicine can improve the E.coli of rats in constipation status, and their effects have no differences;8.In these experimental groups, only acupuncture combined with medicine can improve the bifidobacteria content of fecal when rats are in constipation status;9.In these experimental groups, only simple acupuncture can improve the lactobacillus content of fecal when rats are in constipation status;10.When it comes to gender, only the drug group has differences in fecal water content and 24 hours stool grains per kg body weight,and results show that female is better than male.The reason is probably that female and male rats have different drug resistance.It indicates that we should lower the dose when treat female rats;11.Generally consider all the observation indicators, and clinical medical expenses ,we recommends that: for STC patients, the propriate treatment method is simple acupuncture treatment; acupuncture combined with medicine is suitable for patients with hard stool or less qualitative; When patients’ fecal flora analysis indicates dysbacteriosiscircumstances,they should try to avoid oral cisapride or drugs alike,but can use preparations which containing probiotics.更多还原