CAJ | 학술논문

目的：探讨 SD 大鼠食物过敏动物模型建立的适宜条件及该模型消化系统超微结构的改变。方法30只 SD 大鼠随机分为食物过敏组（20只），生理盐水组（10只）。食物过敏组第1天及第15天以卵清蛋白（ovalbumin，OVA）低剂量腹腔注射进行基础致敏及强化致敏；第20天起以 OVA 进行高剂量隔日灌胃激发至第49天；对照组分别以等量生理盐水进行腹腔注射及灌胃。分别于1、2、3、4、5、6和7周采用酶联免疫吸附试验检测血清 OVA-IgE 抗体水平，同时观察大鼠腹泻情况。第6周后处死动物，取其十二指肠 Treitz 韧带以下5 cm，经固定、包埋、切片，甲苯胺兰染色行肥大细胞完整性评估，HE 染色下观察肠道病理变化，电镜观察肠黏膜超微结构。结果食物过敏组 OVA-IgE 水平在第4～6周逐渐升高，第7周下降，与对照组相比差异有统计学意义（P ＜0．01）。第6周食物过敏组大鼠均出现腹泻。食物过敏组与对照组肠黏膜嗜酸粒细胞密度[（16．48±2．32）个/HPF vs．（1．70±0．42）个/HPF]、肥大细胞数总数[（9．02±1．55）个/HPF vs．（1．84±0．87）个/HPF]和完整性百分率[（36．50％±12．54％）vs．（92．20％±6．87％）]比较，差异有统计学意义（均 P ＜0．01）。电镜和光镜下观察，食物过敏组动物小肠绒毛稀疏、脱落，细胞连接模糊，线粒体广泛肿胀，内质网扩张，内质网脱颗粒。结论采用 SD 大鼠通过低剂量腹腔注射与高剂量灌胃激发至6周可成功建立理想食物过敏动物模型，卵清蛋白致敏大鼠小肠肠黏膜损伤明显，细胞连接模糊，可能在食物过敏发病机制中起重要作用。
목적：탐토 SD 대서식물과민동물모형건립적괄의조건급해모형소화계통초미결구적개변。방법30지 SD 대서수궤분위식물과민조（20지），생리염수조（10지）。식물과민조제1천급제15천이란청단백（ovalbumin，OVA）저제량복강주사진행기출치민급강화치민；제20천기이 OVA 진행고제량격일관위격발지제49천；대조조분별이등량생리염수진행복강주사급관위。분별우1、2、3、4、5、6화7주채용매련면역흡부시험검측혈청 OVA-IgE 항체수평，동시관찰대서복사정황。제6주후처사동물，취기십이지장 Treitz 인대이하5 cm，경고정、포매、절편，갑분알란염색행비대세포완정성평고，HE 염색하관찰장도병리변화，전경관찰장점막초미결구。결과식물과민조 OVA-IgE 수평재제4～6주축점승고，제7주하강，여대조조상비차이유통계학의의（P ＜0．01）。제6주식물과민조대서균출현복사。식물과민조여대조조장점막기산립세포밀도[（16．48±2．32）개/HPF vs．（1．70±0．42）개/HPF]、비대세포수총수[（9．02±1．55）개/HPF vs．（1．84±0．87）개/HPF]화완정성백분솔[（36．50％±12．54％）vs．（92．20％±6．87％）]비교，차이유통계학의의（균 P ＜0．01）。전경화광경하관찰，식물과민조동물소장융모희소、탈락，세포련접모호，선립체엄범종창，내질망확장，내질망탈과립。결론채용 SD 대서통과저제량복강주사여고제량관위격발지6주가성공건립이상식물과민동물모형，란청단백치민대서소장장점막손상명현，세포련접모호，가능재식물과민발병궤제중기중요작용。
Objective To investigate the methodology of establishing food allergy animal model in SD rats and the ultrastructure change of the digestive system.Method 30 SD rats were randomly divided into ovalbumin group(20 rats)and normal saline group(10 rats).Intraperitoneal injection of low dose of sensitization and enhanced sensitization with OVA were performed in OVA group rats on day 1 and day 15 respectively.High-doses intragastric OVA challenge was performed in OVA group rats every two days since day 20 to day 49. Intraperitoneal injection and intragastric administration with same dose normal saline was performed in control group.Serum OVA-IgE antibodies were detected by using ELISA method at week 1,2,3,4,5,6, 7 respectively.Diarrhea was observed among the rats.Rats were sacrificed at week 6,5 cm long duodenum was clipped from ligament of Treitz to distal end.Toluidine blue and HE staining were performed for the mast cell integrity and pathological changes assessment of intestine respectively after fixation,embedding and sectioning.The ultrastructure of intestinal mucosa was observed under transmission electron microscope. Results The OVA-IgE level of OVA group increased gradually from week 4 to 6,but declined at week 7, compared to the normal control group,the difference was statistically significant.20 rats were all attacked by diarrhea at weeks 6.Jejunal mucosal eosinophil density [(16.48 ±2.32)cells/HPF vs.(1.70 ±0.42) cells/HPF]and mast cell number [(9.02 ±1.55)cells/HPF vs.(1.84 ±0.87)cells/HPF]were higher, but the mast cell integrity percentage [(36.50% ±12.54%)vs.(92.20% ±6.87%)]was lower than control group,The difference was statistically significant (P <0.01).The rarefaction and shedding of villi, obscure cell junction,extensive mitochondrial swelling,dilatation and degranulation endoplasmic reticulum was observed in small intestine in OVA group under the light and electron microscope.Conclusions The food allergy SD rats model was established successfully based on low-dose intraperitoneal injection and high-dose intragastric challenge in 6 weeks.Small intestinal mucosal injury obscure cell junction can be observed in OVA sensitized rats,which may play an important role in the pathogenesis of food allergy.