二甲肼和葡聚糖硫酸钠联合建立溃疡性结肠炎癌变小鼠模型

发布时间：2018-04-07 18:10

本文选题：溃疡性结肠炎相关的结肠癌　切入点：小鼠模型　出处：《浙江大学》2005年硕士论文

【摘要】：结肠直肠癌是溃疡性结肠炎最严重的并发症之一,病程较长(8-10年以上),病变范围广泛,或者合并有原发性硬化性胆管炎的溃疡性结肠炎患者并发结肠直肠癌变的危险性大大增加。尽管溃疡性结肠炎所伴发的结肠直肠癌仅占了单纯结肠癌的百分之一,但它却是溃疡性结肠炎患者的主要死亡原因之一。目前我们对溃疡性结肠炎癌变的发病机理认识还不十分清楚。溃疡性结肠炎癌变一般有较长的病史,收集一定数量的病例需要较长时间,因此动物模型是研究溃疡性结肠癌变较为理想的手段。单纯溃疡性结肠炎的动物模型较多,但只有极少数适合于溃疡性结肠炎伴癌变的研究,其中研究较多的是葡聚糖硫酸钠。小鼠自由饮用5%-10%葡聚糖硫酸钠4-9天可致急性结肠炎;饮用3%-5%葡聚糖硫酸钠7天继以普通饮用水7-14天可致慢性结肠炎;该模型在临床表现,病理观察及免疫检查等方面能较好地模拟人类的溃疡性结肠炎。近年来,少数研究者发现小鼠饮用葡聚糖硫酸钠和普通饮用水数个循环,可有不典型增生和癌变的发生,,但所需实验周期较长并且癌变率低。二甲肼是一种特异的结肠致癌物。动物实验表明二甲肼诱发的结肠癌在组织,形态及解剖结构上与人类的大肠肿瘤较为相似。但是二甲肼也只有在大剂量(20-40mg/kg)和长时间(6-12个月)下才能诱发大肠癌。浙江大学硕士学位论文实验方法 1.小鼠实验实验小鼠分A组(实验组)及B一D组(对照组),每组10一12只小鼠;A组Zomg/kg二甲脱腹腔注射后一周给予3%葡聚糖硫酸钠7天继以普通饮用水14天自由饮用,共三个循环;B组生理盐水腹腔注射后给予三个循环的葡聚糖硫酸钠和饮用水;C组和D组各给予ZOmg/kg二甲脐腹腔注射和生理盐水腹腔注射对照。 2.结肠炎临床观察整个实验过程每周测体重;3%葡聚糖硫酸钠饮用3或4天后、饮用结束当天及停用一周后检测粪便隐血;每日观察粪便性状及是否有肉眼血便。 3.病理检测实验小鼠乙醚麻醉处死后分离肛门至回盲部连接处整段大肠。测量大肠长度,垂直剖开后生理盐水冲洗。肉眼观察后予409/L福尔马林缓冲液固定。大肠平分三部分(前,中和后)。每部分5一6片予常规包埋、固定、切片后苏木素一伊红染色进行病理观察。 4.免疫组织化学法检测p一eatenin and ps3常规二步法检测p一eatenin and p53 在不典型增生,癌变及正常组织中的表达情况。结果 1.体重、粪便及大肠长度首次饮用3%葡聚糖硫酸钠的第七天,A、B组中共有75.0%(18/24)小鼠出现腹泻和大便潜血或肉眼血便,这些症状在饮用普通饮用水14天后全部消失。第二次及第三次饮用3%葡聚糖硫酸钠末分别有45.5%(10/22)和36.4%(8/22)出现腹泻或隐血。饮用葡聚糖硫酸钠组的小鼠体重增加缓慢。实验结束时A、B组的平均体重明显低于D组。饮用葡聚糖硫酸钠组小鼠的大肠明显缩短,二甲腆腹腔注射并不影响大肠长度。 2.不典型增生和癌变的发生情况 A组中90.9%(1 0/11),J“鼠至少发生一处不典型增生或/和癌变。A组中有5
[Abstract]:Colorectal cancer is one of the most serious complications of ulcerative colitis, longer duration (8-10 years), a wide range of lesions, or with the risk of primary sclerosing cholangitis in patients with ulcerative colitis complicated with colorectal cancer become greatly increased. In spite of ulcerative colitis and colorectal cancer development only simple colon cancer one percent, it is but one of the main causes of death in patients with ulcerative colitis. We are now on the carcinogenesis of ulcerative colitis pathogenesis is still not clear.
Ulcerative colitis canceration has a long history, collect a certain number of cases requires a longer time, so is the study of animal model of ulcerative colon carcinogenesis is an ideal means. More simple animal model of ulcerative colitis, but only a handful of well suited for the study of ulcerative colitis associated with carcinogenesis, which is mostly studied by dextran sulfate sodium. Mice drank 5%-10% dextran sulfate sodium 4-9 days can cause acute colitis; drinking 3%-5% dextran sulfate sodium for 7 days followed by 7-14 days of ordinary drinking water can cause chronic colitis; the model in clinical manifestation, pathological observation and immunohistochemical examination can simulate human ulcerative colitis. In recent years, some researchers found that the mice drinking dextran sulfate sodium and ordinary drinking water, number of cycles, may have atypical hyperplasia and canceration, but required a longer experimental period and cancer Change rate is low.
Two hydrazine is a colon specific carcinogen. Animal experiments showed that two hydrazine induced colon cancer in tissue morphology and anatomy to human colorectal cancer is similar. But the two methyl hydrazine only in high dose (20-40mg/kg) and long time (6-12 months) to induce colon cancer.
Master Dissertation of Zhejiang University
Experiment method
1. mice of experimental mice were randomly divided into A group (experimental group) and B group (D control group), each group of 10 only a small 12
Rat; group A Zomg/kg two a week off after the intraperitoneal injection of 3% dextran sulfate sodium for 7 days followed by ordinary drink
14 days of free drinking water, a total of three cycles; group B intraperitoneal injection of normal saline after three cycles of Portugal
Chitosan sodium sulfate and drinking water; C group and D group were treated with two ZOmg/kg intraperitoneal injection of saline and a navel
Intraperitoneal injection of control.
Clinical observation of 2. colitis were measured weekly throughout the experiment; 3% dextran sulfate sodium drinking 3 or 4 days
After drinking, the end of the day and a week after discontinuation of fecal occult blood test; daily observation of excrement and whether there is a naked eye
Bloody.
3. pathological examination of experimental mice were sacrificed after ether anesthesia anal separation to the ileocecal junction of entire colon measured.
The amount of large intestine length, normal saline flush vertical open. Naked eye after treated with 409/L buffered formalin fixed
.
The three part (e. split before and after). Each part of a 5 6 pieces of routine embedding, fixed, sliced Su
Lignin staining for pathological observation.
To detect the p p 4. immunohistochemistry eatenin and PS3 eatenin and p53 a conventional two step method
In atypical hyperplasia, the expression of cancerous and normal tissues.
Result
1. weight and length of fecal E.
The first seventh days of drinking 3% dextran sulfate sodium, A, B group, 75% (18/24) mice
Diarrhea and fecal occult blood or gross blood stool, these symptoms disappeared in the drinking of ordinary drinking water for 14 days.
Second times and three times of drinking 3% dextran sulfate sodium respectively at the end of 45.5% (10/22) and 36.4% (8/22) appears
Diarrhea or occult blood.
The body weight of mice drinking dextran sulfate sodium group increased slowly. At the end of the experiment of A, the average weight of B group
Significantly lower than D group.
Drinking dextran sulfate sodium group E. significantly shortened two a shy intraperitoneal injection does not affect the length of E.
Degrees.
2. cases of atypical hyperplasia and canceration
In 90.9% A (1 0/11), J rats occurred at least an atypical hyperplasia or / and canceration in 5 of.A group

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R-332

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