發(fā)布時(shí)間：2018-05-12 11:06

  本文選題：陽明腑實(shí)證 + 大鼠模型��； 參考：《大連醫(yī)科大學(xué)》2009年碩士論文

【摘要】： 背景:《傷寒論》是中醫(yī)學(xué)的經(jīng)典論著,其對(duì)臨床辨證用藥具有重大的指導(dǎo)作用。陽明腑實(shí)證是許多外感熱病病程中所出現(xiàn)的邪熱內(nèi)熾、又伴有腹部實(shí)證癥狀的一組全身性綜合證候。它以痞、滿、躁、實(shí)、堅(jiān)為主癥,可以出現(xiàn)在許多疾病過程中,例如:嚴(yán)重腹內(nèi)感染、重型急腹癥、嚴(yán)重創(chuàng)傷、大面積燒傷、大手術(shù)后等。具有發(fā)病急、病情重、變化快、并發(fā)癥多的特點(diǎn),如不及時(shí)診治�？梢鹦菘�、DIC、ARDS甚至多器官功能衰竭(MODS)從而危及生命。但其辨證缺乏客觀性,療效缺乏客觀評(píng)價(jià)指標(biāo),因此,在進(jìn)行文獻(xiàn)整理與臨床驗(yàn)證等研究的同時(shí),吸收現(xiàn)代科學(xué)知識(shí)的方法,加強(qiáng)實(shí)驗(yàn)研究是十分必要的。在醫(yī)學(xué)研究中,建立疾病的動(dòng)物模型是一個(gè)重要的手段。在陽明腑實(shí)證的研究中,復(fù)制動(dòng)物模型的方法也是必不可少的。特別是近年來,建立大鼠陽明腑實(shí)證動(dòng)物模型是當(dāng)務(wù)之急。有了合適的動(dòng)物模型,才能建立進(jìn)一步進(jìn)行中西醫(yī)結(jié)合診治的平臺(tái)。 目的:采用灌服熱性中藥與自身糞便相結(jié)合的方法制備大鼠陽明腑實(shí)證模型,并與已得到廣泛認(rèn)可的兩種模型(盲腸結(jié)扎穿孔法、腹腔注射酵母多糖與液體石蠟法)對(duì)大鼠血漿ET、腫瘤壞死因子(TNF-α)、IL-10含量進(jìn)行定量測(cè)定。用間苯三酚法檢測(cè)各組大鼠小腸對(duì)D-木糖的吸收功能,并觀察肺、小腸、肝、腎組織的病理形態(tài)學(xué)變化。 方法:SD大鼠40只,雌雄各半,體重180~220g。將實(shí)驗(yàn)大鼠隨機(jī)分成4組,每組10只。對(duì)照組、灌服熱性中藥與自身糞便組(簡(jiǎn)稱中藥組)、盲腸結(jié)扎穿孔組(簡(jiǎn)稱結(jié)扎組)、腹腔注射ZymosanA與無菌液體石蠟混懸液組(簡(jiǎn)稱注射組)。中藥組:以附子、吳茱萸、干姜以1∶1∶1作水煎劑,20g/kg灌服12天后,禁食,100g/L自身糞便混懸液2ml灌胃,每天2次,連續(xù)2天,同時(shí)5%乙醇代水隨意飲用,每日更換,保證濃度。結(jié)扎組:實(shí)驗(yàn)動(dòng)物正常飼養(yǎng),每天灌服自來水20g/kg體重,連續(xù)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,結(jié)扎盲腸根部,于距結(jié)扎線約0.5cm處以5ml注射器針頭穿孔并縫合切口,24h后取材。其他組僅開腹游離盲腸不做結(jié)扎穿孔后縫合切口。注射組:實(shí)驗(yàn)動(dòng)物正常飼養(yǎng),每天灌服自來水20g/kg體重,連續(xù)14天。第15日以無菌ZymosanA粉劑與無菌液體石蠟制成混懸液,按1.0mg/g體重腹腔注射,24小時(shí)后取材,其他各組注射等量無菌液體石蠟。血漿D-木糖測(cè)定(間苯三酚法):各組大鼠于處死前一小時(shí)先灌服50g/lD-木糖溶液0.3g/kg,1h后腹主動(dòng)脈取血,制備血漿,用間苯三酚法進(jìn)行檢測(cè)。血漿ET、腫瘤壞死因子(TNF-α)、IL-10均采用ELISA法檢測(cè),下腔靜脈取血,制備血漿嚴(yán)格按試劑盒說明操作。處死大鼠后,取出肝、肺、腎、小腸,用生理鹽水沖洗干凈,制作病理切片,HE染色,光鏡觀察。 結(jié)果: 1.一般狀況:中藥組大鼠灌服熱性中藥3天后出現(xiàn)體重降低,豎毛,活動(dòng)增加,飲水量增加,尿量減少并發(fā)黃;灌自身糞便當(dāng)天出現(xiàn)排便時(shí)間延長(zhǎng),排便粒數(shù)減少,大便干結(jié),并成圓珠狀或串珠狀;其他組大鼠第15日前飲食,大便正常,毛發(fā)有色澤、行為狀態(tài)正常。 2.血漿ET含量:與對(duì)照組相比較,三個(gè)模型組的血漿ET含量均明顯升高,差異具有顯著性(P0.05),三個(gè)模型組之間差異沒有顯著性(P0.05)。 3.血漿TNF-α含量:與對(duì)照組相比較,三個(gè)模型組的血漿TNF-α含量均明顯升高,差異具有顯著性(P0.05),三個(gè)模型組之間差異沒有顯著性(P0.05)。 4.血漿IL-10含量:與對(duì)照組相比較,三個(gè)模型組的血漿IL-10含量均明顯升高,差異具有顯著性(P0.05),三個(gè)模型組之間差異沒有顯著性(P0.05)。 5.D-木糖吸收率:與對(duì)照組相比較,三個(gè)模型組的血漿D-木糖含量均明顯升高,差異具有顯著性(P0.05),三個(gè)模型組之間差異沒有顯著性(P0.05)。 6.各組大鼠組織病理形態(tài)學(xué)變化:三個(gè)模型組大鼠的主要臟器(小腸、肝、肺、腎)均表現(xiàn)為明顯的炎性病理改變。 結(jié)論: 1.灌服熱性中藥與自身糞便法在各個(gè)檢測(cè)指標(biāo)上均達(dá)到以上提及的兩種造模方法的檢測(cè)結(jié)果,其造模原理更加符合中醫(yī)理論。 2.血漿ET、腫瘤壞死因子(TNF-α)、IL-10含量、D-木糖的吸收率;以及肺、肝、腎、小腸的部分組織病理形態(tài)學(xué)改變可作為此證的客觀診斷指標(biāo)。
[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.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R228;R-332

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