【作者】 陈龙明；

【导师】 李群伟；

【作者基本信息】 泰山医学院 ， 流行病与卫生统计学， 2008， 硕士

【摘要】 目的:利用亚慢性动物实验观察脱氧雪腐镰刀菌烯醇(DON)、N-亚硝基化合物(NOC)对实验动物的致病作用,主要观察对动物消化系统的影响,探讨DON、NOC对实验动物食管、胃、肝脏、直肠组织急慢性损伤及其潜在的致癌作用,为消化系统肿瘤的病因预防提供理论依据。方法:以断乳7日的小鼠为实验对象,雌雄各半,随机分为四组,每组32只,其中DON组染毒剂量为DON 0.5mg/kg/次,NOC组染毒剂量为NOC 0.25mg/kg/次,DON+ NOC组染毒剂量为DON0.5mg/kg/次+NOC0.25mg/kg/次,染毒方式为隔日灌胃,对照组用等体积生理盐水灌胃。实验周期满60日后处死小鼠,取小鼠食管、胃、肝脏、直肠组织制作HE切片,观察各实验组、对照组食管、胃、肝脏、直肠组织癌前病变、急慢性损伤。此外,对泰安地区粮食样品中DON进行抽样检测,观察DON含量与肿瘤关系。利用SAS9.0统计软件对数据进行处理、分析。结果:1.对食管组织的影响:实验组出现了乳头延长、上皮乳头状增生、上皮细胞空泡化、上皮增生角化不全改变,阳性率分别为:34.04%、8.51%、8.51%、20.21%;其中DON+NOC实验组乳头延长、上皮乳头状增生、上皮增生角化不全改变阳性率较高,分别为:50.00%、15.63%、25.00%;DON组乳头延长阳性率较高,为32.26%;NOC组乳头延长、上皮增生角化不全改变阳性率较高,分别为:19.35%、25.81%,对照组未出现上述改变。实验组乳头延长、上皮增生角化不全改变与对照组进行χ2检验,有统计学意义(P<0.05)。2.对胃组织的影响:实验组胃出现息肉形成趋势、腺体轻度异型、深层上皮轻度异型、黏膜上皮坏死、腺体坏死、上皮增生、脉管扩张等组织学改变,阳性率分别为:31.91%、29.79%、42.55%、40.43%、52.13%、13.83%、9.57%;其中DON+NOC实验组息肉形成趋势、深层上皮轻度异型、黏膜上皮坏死阳性率高,分别为:46.88%、62.50%、59.38%;DON组息肉形成趋势、腺体轻度异型、腺体坏死和黏膜上皮坏死阳性率较高,分别为:22.58%、29.03%、25.81%和45.16%;NOC组深层上皮轻度异型、黏膜上皮坏死和腺体坏死阳性率较高,分别为:38.71%、51.61%和52.61%;各组均有炎症性改变。实验组小鼠出现的胃息肉形成趋势、胃腺体轻度异型、深层上皮轻度异型、腺体坏死和黏膜上皮坏死改变与对照组进行χ2检验,有统计学意义(P<0.05),DON+NOC组与DON组的深层上皮轻度异型出现率进行四格表χ2检验,有统计学意义(P<0.007)。3.对肝脏组织的影响:实验组肝脏出现大片坏死、大片脂肪变性、小片脂肪变性、灶性坏死、嗜酸性变、小灶区异型、炎细胞浸润、双核肝细胞、脉管扩张病变,阳性率分别为:7.45%、39.36%、22.34%、13.83%、17.02%、17.02%、76.60%、50.00%、31.91%;其中DON+NOC组组织大片坏死、大片脂肪变性、灶性坏死、双核肝细胞和灶区异型阳性率高,分别为:21.88%、65.63%、28.13%、25.00%和25.00%;DON组大片脂肪变性和小灶区异型阳性率较高,分别为:22.58%和25.81%;NOC组大片脂肪变性和脉管扩张阳性率较高,分别为:29.03%和25.58%;对照组肝脏组织出现了脉管扩张、小片脂肪变性、炎细胞浸润、嗜酸性变改变,阳性率分别为:6.25%、75.00%、45.45%、33.33%。实验组肝脏出现大片坏死、大片脂肪变性、灶性坏死、小灶区异型、双核肝细胞、脉管扩张各病变与对照组进行χ2检验,均有统计学意义(P<0.05),DON+NOC组分别与DON组、NOC组出现的大片脂肪变性进行四格表χ2检验,有统计学意义(P<0.007)。4.对直肠组织的影响:实验组出现腺体轻度异型、黏膜上皮坏死、固有膜炎细胞、黏膜水肿、腺体结构紊乱、淋巴滤泡形成、嗜酸粒细胞等改变,阳性率分别为:40.43%、43.62%、79.79%、58.51%、14.89%、8.51%、23.40%;其中DON+NOC组腺体轻度异型、黏膜上皮坏死和腺体结构紊乱阳性率高,分别为:71.88%、46.88%和18.75%;DON组腺体轻度异型、黏膜上皮坏死阳性率较高,分别为:29.03%、32.26%;NOC组腺体轻度异型、黏膜上皮坏死和腺体结构紊乱阳性率较高,分别为:19.35%、51.61%和16.13%;对照组组织出现了炎症水肿、淋巴滤泡和嗜酸粒细胞浸润改变,阳性率分别为:25.00%、3.13%、3.13%。实验组小鼠直肠组织出现腺体轻度异型、黏膜上皮坏死脱落、腺体结构紊乱、嗜酸粒细胞、固有膜炎细胞改变与对照组进行χ2检验,有统计学意义(P<0.05),DON+NOC组分别与DON组、NOC组出现的直肠腺体轻度异型进行四格表χ2检验,有统计学意义(P<0.007)。5.泰安市食管癌高发区小麦面、玉米面样品中DON含量与低发区进行t检验,有统计学意义(p<0.05)。结论:1.DON可诱发消化系统出现组织坏死、炎性水肿、大片脂肪变性等急慢性病理损伤和息肉形成趋势、腺体上皮异型增生等癌前病变。2.NOC可引起实验动物出现组织坏死、水肿变性、脂肪变性等急慢性病理损伤和息肉形成趋势、腺体上皮轻度异型、食管乳头延长等癌前病变。3.DON+NOC组实验动物出现消化系统组织坏死、脂肪变性、炎性水肿等急慢性病理损伤,导致息肉形成趋势和腺体、上皮异型增生等癌前病变,且病变程度强于DON、NOC单独作用。4.泰安地区食管癌高发区粮食样品中DON含量高于低发区。5.实验结果表明:DON、NOC对消化系统具有明显的损伤,同时具有潜在的致癌效应,且二者间存在协同作用。更多还原

【Abstract】 Objective:We carried out the subchronic animal experiments to oberserve morbigenous effects,which were caused by Dexoynivalenol(DON) and Nitrogen-nitroso compound (NOC).Our study mostly aimed to elucidate the effects they had on the digestive systems of the experimental animals and the effects of acute, chronic toxicity, as well as potential carcinogenic effects on the organs or tissues,such as oesophagus,stomach,liver and rectum of the experimental ainimals.This work might provide a basis theory for preventing digestive system carcinorma.Method:Experimental mice weaned 7 days were divided into four groups at random, there were 32 mice in each group,with females and males accounting for one half.Every mouse in DON group was given DON with the oral dose of 0.5mg/kg body weight a time, every mouse in NOC group was given NOC with the oral dose of 0.25 mg/kg body weight a time,every mouse in DON + NOC group was given DON with the oral dose of 0.5mg/kg and NOC with the oral dose of 0.25mg/kg body weight a time,control group was given isometrical physiological saline,two days a time.Sixty days later,all the mice were killed and the organs or tissues of oesophagus,stumach,liver and rectum were abtained.HE pathological slices of this organs were observed.In dddition,the concentration of DON in grain sanples in TaiAn was detected using HPLC-MS in order to explain the relationship between DON and the tumour.All the statistical analyses performed by the software of SAS9.0.Result:1.Poisonous effect on oesophagus tissue:Pathological changes of the oesophagus tissues were observed in the animals of the experimental groups,and the positive ratios in mammilla extension,the epithelium tissue corpora mammillaria accrementition,cellula epithelia vacuolization,accrementition of epithelium tissue parakeratosis were 34.04%,8.51%,8.51% and20.21%,respectively.The DON+NOC group had high positive ratio in mammilla extension, the epithelium tissue corpora mammillaria accrementition,accrementition of epithelium tissue parakeratosis,with 50.00%、15.63%、25.00%,respectively.The DON group had higher positive ratio in mammilla extension,with 32.26%.The NOC group had higher positive ratios in mammilla extension and accrementition of epithelium tissue parakeratosis,and the positive ratio was 19.35%、25.81%,respectively.However,the above changes were not observed in the control group.The difference in mammilla extension and accrementition of epithelium tissue parakeratosis were statistically significant between the experimental group and control group (P﹤0.05) according toχ2 test.2.Poisonous effect on stomach tissue:Pathological changes of stomach tissues,were also found in the experimental animals,the positive ratios in the polyp formation tendency,the gland mild heterotype,the epithelium tissue in deep lamella mild heterotype,the epidermis mucosae necrosis,necrosis of the gland,accrementition of epithelium tissue,and the Vessel expansion were 31.91%,29.79%,42.55%,40.43%,52.13%,13.83%,9.57%,respectively.The DON+NOC group had higher positive ratios in the polyp formation tendency,the epithelium tissue in deep lamella mild heterotype and the epidermis mucosae necrosis,the positive ratios were 46.88%、62.50%、59.38%, respectively.The DON group had higher positive ratios in the polyp formation tendency,the epithelium tissue in deep lamella mild heterotype,the epidermis mucosae necrosis, necrosis of the gland,and the positive ratios was 22.58%、29.03%、25.81% and 45.16%,respectively.The NOC group had higher positive ratios in the epithelium tissue in deep lamella mild heterotype,the epidermis mucosae necrosis,the necrosis of the gland,and the positive ratio was 38.71%、51.61% and 52.61%,respectively.Inflammation change was detected in every group.The difference in the polyp formation tendency,the epithelium tissue in deep lamella mild heterotype,the epidermis mucosae necrosis,the necrosis of the gland,the gland mild heterotype were statistically significant between experimental groups and control group(P﹤0.05)according toχ2 test.The difference in the epithelium tissue in deep lamella mild heterotype were statistically significant between the DON+NOC group and DON group (P﹤0.007).3.Poisonnous effect on liver tissue:Pathological changes of liver tissues were observed in the experimental groups,and the positive ratio in the large flap necrosis,the large flap fatty degeneration,the scrap fatty degeneration,the focal necrosis,the acidophilia changes,the focal heterotype,the inflammatory cell infiltration,the new born conjugate nuclear liver cell and the vessel expansion pathological changes were 7.45%,39.36%,22.34%,13.83%, 17.02%,17.02%, 76.60%,50%,31.91%,respectively.The DON+NOC group had higher positive ratios in the large flap necrosis(21.88%),the large flap fatty degeneration (65.63%),the focal necrosis (28.13%),the focal heterotype(25.00%),the new born conjugate nuclear liver cell(25.00%). The DON group had higher positive ratios in the large flap fatty degeneration and the focal heterotype,and the positive ratio was 22.58% and 25.81%,respectively.The NOC group had higher positive ratios in the large flap fatty degeneration pathological changes and the vessel expansion,and the positive ratios were 29.03% and 25.58% , respectively.However, pathological changes were also detected in the liver tissues in control group,and the positive ratio in the vessel expansion,the scrap fatty degeneration,the acidophilia changes,and inflammatory cell infiltration were 6.25%、75.00%、33.33% and 45.45%,respectively.The difference in the large flap necrosis,the large flap fatty degeneration,the focal necrosis,the focal heterotype,the new born conjugate nuclear liver cell and the vessel expansion pathological changes were statistically significant between the experimental groups and control group(P﹤0.05).The significant differences in the large flap fatty degeneration were also found between the DON+NOC group and control group(P﹤0.007).4.Poisonous effect on rectum tissue:Pathological changes of rectum organ tissues were observed in the experimental groups,and the positive ratios in the gland mild heterotype,the epithelium mucosae necrosis,the membrana propria inflammatory cell infiltration,the mucosae dropsy,the gland disorganization,the folliculus lymphaticus formation and the eosinophilic granulocyte infiltration pathological changes were 40.43%,43.62%,58.51%, 14.89%,8.51%, 23.40%,respectively.The DON+NOC group had higher positive ratios in the gland mild heterotype,the epithelium mucosae necrosis and the gland disorganization,and the positive ratios were 71.88%、46.88% and 18.75%,respectively.The DON group had higher positive ratios in the gland mild heterotype and the epithelium mucosae necrosis,and the positive ratioswere 9.03% and 32.26%,respectively.The NOC group had higher positive ratios in the gland mild heterotype,the epithelium mucosae necrosis and the gland disorganization,and the positive ratios were 19.35%、51.61% and 16.13%,respectively.Pathological changes were also found in control group and the positive ratios in the mucosae dropsy,the folliculus lymphaticus formation and the eosinophilic granulocyte infiltration pathological changes were 25.00%,3.13% and 3.13%.The differences in the gland mild heterotype,the gland disorganization,the epithelium mucosae necrosis,the eosinophilic granulocyte infiltration and the membrana propria inflammatory cell infiltration were statistically significant between the experimental groups and control group (P﹤0.05).In addition,the difference in the gland mild heterotype was also statistically significant between the DON+NOC and DON group(P﹤0.05).Furthermore,the difference in the gland mild heterotype was significant between the DON+NOC group and NOC group. 5.In addition,we detected the DON concentrations in the grain samples in TaiAn,the differences were statistically significant between the areas of higher incidence of oesophageal cancer and those with low incidence(P﹤0.05) .Conclusion:1.DON can induce precancerosis of the digestive system,such as the polyp formation tendency,the gland and epidermis mild heterotype,as well as acute and chronic pathology changes,such as the organ tissue necrosis,the inflammation dropsy and fatty degeneration,etc.2.NOC can also induce precancerosis of the digestive system,such as the polyp formation tendency and the gland and mammilla extension,as well as acute and chronic pathology changes,such as the organization necrosis,the inflammation dropsy denaturaation,big piece fatty degeneration,etc.3.The synergy of DON and NOC may cause digestive system acute and chronic pathology damages,such as,the appearances of necrosis , fatty degeneration and inflammation dropsy,etc.Moerever they also induce precancerosis,such as,the polyp formation tendency and the gland,epidermis heterotype.The above pathological changes were more obvious than DON or NOC were given alone.4.The concentration of DON were higher in grains samples collected from areas with higher incidence of oesophagus than those with lower incidence in Taian.5.Our results indicated:DON,NOC obvious damaged the digestive system.At the same time,they had the latent carcinogenic effects,and there effects obvious synergism.更多还原