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幽门螺杆菌ICR小鼠脾胃湿热证动物模型的建立
胡锦洋,蒋士生,肖梅英
0
(湖南省中医药研究院附属医院,湖南 长沙,410006;湖南省中医药研究院,湖南 长沙,410006;湖南省中医药研究院中药研究所,湖南 长沙,410013)
摘要:
目的:建立脾胃湿热型幽门螺杆菌(Hp)相关性胃炎小鼠模型,评价其胃黏膜病理变化。方法:选择40只ICR小鼠适应性饲养,编号后随机分为空白组10只和造模组30只,空白组正常饲养,不予接种Hp,每天仅以0.9%氯化钠注射液灌胃。将造模组的30只小鼠随机分为3组(M0组、M4组、M8组),M4、M8组小鼠经禁食24h后予50%乙醇0.1ml灌注于每只小鼠胃内,1h后再灌注Hp菌株0.5ml;隔天1次,共灌胃5次(灌胃前6h及灌胃后30min内禁食、禁水)。末次灌胃2h后,随机抽取2只ICR小鼠进行尿素酶试验及胃黏膜细菌培养。细菌培养镜检发现G-弧形杆菌,尿素酶试验阳性,提示Hp相关性胃炎小鼠模型造模成功。2组小鼠再予湿热环境+高脂高糖饮食多种因素综合作用,制备成脾胃湿热型Hp相关性胃炎小鼠模型,继续饲养4周(M4组)、8周(M8组),定期观察动物体质量、症状积分,分别于第4、8周每组各处死1/2动物,取鼠胃腺体组织观察胃炎程度及Hp感染程度。结果:与空白组比较,第4、8周模型组症状积分、病理积分均增加,差异均有统计学意义(P<0.05);与空白组比较,模型组Hp感染的动物数明显增加;与M0组比较,M8组小鼠胃组织Hp感染严重程度显著增加,差异均有统计学意义(P<0.05或P<0.01)。结论:通过灌胃给菌的方法可以成功将Hp定植于小鼠体内,并在现有条件下成功建立脾胃湿热证Hp相关性胃炎小鼠模型,其保存期有8周,且8周模型胃炎程度及Hp感染程度更深。
关键词:  Hp相关性胃炎  脾胃湿热型  小鼠模型  实验研究
DOI:
Establishment of an animal model of Helicobacter pylori-related gastritis with spleen-stomach damp-heat using Institute of Cancer Research mice
HU Jinyang,JIANG Shisheng,XIAO Meiying
(The Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006,Hunan,China;Institute of Chinese Materia Medica,Hunan Academy of Chinese Medicine,Changsha 410013,Hunan,China)
Abstract:
Objective:To establish a mouse model of Helicobacter pylori (Hp)-related gastritis with spleen-stomach damp-heat,and to investigate the pathological changes of gastric mucosa.Methods:A total of 40 Institute of Cancer Research (ICR) mice were given adaptive feeding,and then they were numbered and randomly divided into blank group with 10 mice and model group with 30 mice.The mice in the blank group were given normal feeding and 0.9% sodium chloride injection by gavage without Hp inoculation.The model group was further divided into M0,M4,and M8 groups;the mice in the M4 and M8 groups were given 0.1mL 50% alcohol by gavage after 24-hour fasting,followed by 0.5mL Hp strain by gavage 1 hour later,once every other day for a total of 5 times,with fasting and water deprivation at 6 hours before gavage and 30 minutes after gavage.At 2 hours after the last gavage,2 ICR mice were randomly selected for urease test and bacterial culture of gastric mucosa.Bacterial culture and microscopy found G-campylobacter and urease test yielded positive results,suggesting that a mouse model of Hp-related gastritis was successfully established.Various factors including damp-heat environment and high-fat high-glucose diet were used to establish a mouse model of Hp-related gastritis with spleen-stomach damp-heat.The mice in the M4 and M8 groups were fed for another 4 and 8 weeks,respectively.Body weight and symptom score were observed regularly.Half of the animals were sacrificed at week 4,and the other half were sacrificed at week 8;gastric gland tissue was collected to observe the severity of gastritis and Hp infection.Results:Compared with the blank group,the model group had significant increases in symptom score and pathological score at weeks 4 and 8 (P<0.05).Compared with the blank group,the model group had a significant increase in the number of mice with Hp infection.Compared with the M0 group,the M8 group had a significant increase in the severity of Hp infection in gastric tissue (P<0.05 or P<0.01).Conclusion:The method of giving Hp by gavage can achieve successful colonization of Hp in mice and help to successfully establish a mouse model of Hp-related gastritis with spleen-stomach damp-heat in existing conditions.The model can be preserved for 8 weeks,and there are increases in the severity of gastritis and Hp infection at week 8.
Key words:  Helicobacter pylori-related gastritis  spleen-stomach damp-heat  mouse model  experimental study

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