阳明腑实证大鼠模型的制备及其病理机制研究

发布时间：2018-05-12 11:06

本文选题：阳明腑实证 + 大鼠模型　；参考：《大连医科大学》2009年硕士论文

【摘要】： 背景:《伤寒论》是中医学的经典论著,其对临床辨证用药具有重大的指导作用。阳明腑实证是许多外感热病病程中所出现的邪热内炽、又伴有腹部实证症状的一组全身性综合证候。它以痞、满、躁、实、坚为主症,可以出现在许多疾病过程中,例如:严重腹内感染、重型急腹症、严重创伤、大面积烧伤、大手术后等。具有发病急、病情重、变化快、并发症多的特点,如不及时诊治常可引起休克、DIC、ARDS甚至多器官功能衰竭(MODS)从而危及生命。但其辨证缺乏客观性,疗效缺乏客观评价指标,因此,在进行文献整理与临床验证等研究的同时,吸收现代科学知识的方法,加强实验研究是十分必要的。在医学研究中,建立疾病的动物模型是一个重要的手段。在阳明腑实证的研究中,复制动物模型的方法也是必不可少的。特别是近年来,建立大鼠阳明腑实证动物模型是当务之急。有了合适的动物模型,才能建立进一步进行中西医结合诊治的平台。目的:采用灌服热性中药与自身粪便相结合的方法制备大鼠阳明腑实证模型,并与已得到广泛认可的两种模型(盲肠结扎穿孔法、腹腔注射酵母多糖与液体石蜡法)对大鼠血浆ET、肿瘤坏死因子(TNF-α)、IL-10含量进行定量测定。用间苯三酚法检测各组大鼠小肠对D-木糖的吸收功能,并观察肺、小肠、肝、肾组织的病理形态学变化。方法:SD大鼠40只,雌雄各半,体重180~220g。将实验大鼠随机分成4组,每组10只。对照组、灌服热性中药与自身粪便组(简称中药组)、盲肠结扎穿孔组(简称结扎组)、腹腔注射ZymosanA与无菌液体石蜡混悬液组(简称注射组)。中药组:以附子、吴茱萸、干姜以1∶1∶1作水煎剂,20g/kg灌服12天后,禁食,100g/L自身粪便混悬液2ml灌胃,每天2次,连续2天,同时5%乙醇代水随意饮用,每日更换,保证浓度。结扎组:实验动物正常饲养,每天灌服自来水20g/kg体重,连续14天。第15日以10%水合氯醛腹腔注射麻醉后,取右下腹斜切口入腹,结扎盲肠根部,于距结扎线约0.5cm处以5ml注射器针头穿孔并缝合切口,24h后取材。其他组仅开腹游离盲肠不做结扎穿孔后缝合切口。注射组:实验动物正常饲养,每天灌服自来水20g/kg体重,连续14天。第15日以无菌ZymosanA粉剂与无菌液体石蜡制成混悬液,按1.0mg/g体重腹腔注射,24小时后取材,其他各组注射等量无菌液体石蜡。血浆D-木糖测定(间苯三酚法):各组大鼠于处死前一小时先灌服50g/lD-木糖溶液0.3g/kg,1h后腹主动脉取血,制备血浆,用间苯三酚法进行检测。血浆ET、肿瘤坏死因子(TNF-α)、IL-10均采用ELISA法检测,下腔静脉取血,制备血浆严格按试剂盒说明操作。处死大鼠后,取出肝、肺、肾、小肠,用生理盐水冲洗干净,制作病理切片,HE染色,光镜观察。结果: 1.一般状况:中药组大鼠灌服热性中药3天后出现体重降低,竖毛,活动增加,饮水量增加,尿量减少并发黄;灌自身粪便当天出现排便时间延长,排便粒数减少,大便干结,并成圆珠状或串珠状;其他组大鼠第15日前饮食,大便正常,毛发有色泽、行为状态正常。 2.血浆ET含量:与对照组相比较,三个模型组的血浆ET含量均明显升高,差异具有显著性(P0.05),三个模型组之间差异没有显著性(P0.05)。 3.血浆TNF-α含量:与对照组相比较,三个模型组的血浆TNF-α含量均明显升高,差异具有显著性(P0.05),三个模型组之间差异没有显著性(P0.05)。 4.血浆IL-10含量:与对照组相比较,三个模型组的血浆IL-10含量均明显升高,差异具有显著性(P0.05),三个模型组之间差异没有显著性(P0.05)。 5.D-木糖吸收率:与对照组相比较,三个模型组的血浆D-木糖含量均明显升高,差异具有显著性(P0.05),三个模型组之间差异没有显著性(P0.05)。 6.各组大鼠组织病理形态学变化:三个模型组大鼠的主要脏器(小肠、肝、肺、肾)均表现为明显的炎性病理改变。结论: 1.灌服热性中药与自身粪便法在各个检测指标上均达到以上提及的两种造模方法的检测结果,其造模原理更加符合中医理论。 2.血浆ET、肿瘤坏死因子(TNF-α)、IL-10含量、D-木糖的吸收率;以及肺、肝、肾、小肠的部分组织病理形态学改变可作为此证的客观诊断指标。
[Abstract]:Background: "the theory of typhoid fever" is a classic book of traditional Chinese medicine. It has a great guiding role in the clinical syndrome differentiation. For example, severe abdominal infection, severe acute abdomen, severe trauma, large area burns, large operation, and so on. It has the characteristics of acute disease, heavy disease, rapid change, and many complications, such as not timely diagnosis and treatment often can cause shock, DIC, ARDS even multiple organ failure (MODS) endangers life. But the syndrome differentiation lacks objectivity and the effect lacks objective evaluation. Therefore, it is necessary to absorb modern scientific knowledge and strengthen experimental research. In medical research, it is an important means to establish animal model of disease in medical research. In the study of Yangming Fu, the method of reproducing animal models is also essential. In recent years, it is urgent to establish an empirical animal model of Yangming Fu organs in rats. With appropriate animal models, a platform for further diagnosis and treatment of Chinese and Western medicine can be established.
Objective: to prepare an empirical model of rat Yangming Fu Fu by the combination of hot Chinese medicine and self feces, and the quantitative determination of ET, tumor necrosis factor (TNF- alpha) and IL-10 in rat plasma with two widely recognized models (cecum ligation perforation method, intraperitoneal injection of yeast polysaccharide and liquid paraffin). The absorption of D- xylose was detected in small intestine of rats, and the pathomorphological changes in lung, small intestine, liver and kidney were observed.
Methods: 40 SD rats were divided into 4 groups of male and male rats and male and female. The experimental rats were randomly divided into 4 groups, 10 rats in each group. The control group was given the thermal traditional Chinese medicine and the self fecal group (Chinese medicine group), the cecum ligation group (ligation group), the abdominal injection of ZymosanA and the aseptic liquid paraffin suspension group (abbreviated as the injection group). Dry ginger used 1: 1: 1 as water decoction, 20g/kg for 12 days, fasting, 100g/L self faecal suspension 2ml gavage, 2 times a day for 2 days, and 5% ethanol generation at the same time drinking, daily replacement, to ensure the concentration. The experimental animals were fed normal feeding, daily 20g/kg weight of tap water, for 14 days. 10% hydrate chloral intraperitoneal injection anesthesia on the fifteenth day. After drunken, take the right lower abdomen oblique incision into the abdomen, ligate the caecum root, and put the needle piercing of the 5ml syringe needle and suture the incision about 0.5cm from the ligation line, and then pick up the incision after 24h. The other groups only open the open caecum and do not do the suture incision after the ligation and perforation. The injection group: the experimental animals were fed normally, and the 20g/kg weight of the tap water was taken for 14 days. The sterile Z was used for fifteenth days. YmosanA powder and aseptic liquid paraffin were made into a suspension and injected into the abdominal cavity for 24 hours after 24 hours of injection. The other groups were injected with the same amount of aseptic liquid paraffin. Plasma D- xylose determination (interphenolol method). The rats were given 50g/lD- xylose solution 0.3g /kg first an hour before death, and blood was taken from the abdominal aorta after 1h, and the plasma was prepared with phenolol three. The method was tested. The plasma ET, tumor necrosis factor (TNF- alpha), IL-10 were detected by ELISA, the blood was taken from the inferior vena cava, and the preparation of plasma was strictly controlled by the kit. After the death of the rats, the liver, lung, kidney and small intestine were removed, and the normal saline was washed, pathological section, HE staining, and light microscopy were made.
Result:
1. general condition: the rats in the traditional Chinese medicine group were given the weight loss, the vertical hair, the increase of the activity, the increase of the drinking water, the decrease of the urine and the yellowing of the Chinese herbal medicine for 3 days. The time of defecation was prolonged, the number of defecation was reduced, the stool was reduced, and the stool was formed into a round or beaded shape. The other groups of rats had a diet fifteenth days ago, the stool normal, and hairs color and lustre. The behavior is normal.
2. plasma ET content: compared with the control group, the plasma ET content in the three model groups increased significantly (P0.05), and there was no significant difference between the three model groups (P0.05).
3. plasma TNF- alpha content: compared with the control group, the plasma TNF- alpha content in the three model groups increased significantly (P0.05), and there was no significant difference between the three model groups (P0.05).
4. plasma IL-10 content: compared with the control group, the plasma IL-10 content in the three model groups increased significantly (P0.05), and there was no significant difference between the three model groups (P0.05).
5.D- xylose absorption rate: compared with the control group, the plasma D- xylose content of the three model groups increased significantly (P0.05), and there was no significant difference between the three model groups (P0.05).
6. histopathological changes of rats in each group: the main organs (small intestine, liver, lung and kidney) of three model groups showed obvious inflammatory pathological changes.
Conclusion:
1. the test results of two kinds of modeling methods mentioned above were reached on each test index, which were more in line with the theory of traditional Chinese medicine.
2. plasma ET, tumor necrosis factor (TNF- alpha), IL-10 content, absorption of D- xylose, and partial histopathological changes in lung, liver, kidney and small intestine can be used as an objective diagnostic indicator of this syndrome.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R228;R-332

【参考文献】