摘要
建立轮状病毒(rotavirus,RV) SA-11株Balb/c乳鼠动物模型,研究其发病机制,为后续疫苗保护性评价奠定基础。用不同剂量的SA11毒株单次灌胃感染7 d龄的Balb/c乳鼠,于感染后不同时间点观察其腹泻、活动能力及精神状态等生理指标,并解剖乳鼠,取其心、肝、脾、肺、小肠及肾等组织。免疫组化和免疫荧光检测轮状病毒抗原分布,HE染色观察小肠形态学改变,组织细胞原位凋亡检测观察攻毒后小肠绒毛细胞的凋亡情况。研究发现,高剂量和中剂量攻毒组在攻毒24 h后全组乳鼠即可发生明显腹泻,对照组和低剂量组乳鼠无腹泻发生。攻毒72 h后解剖乳鼠,高、中剂量攻毒组的乳鼠肠道可见明显充血及肠胀气,病理切片HE染色显示小肠绒毛发生大量空泡变性及小肠绒毛顶端破损,小肠绒毛原位凋亡检测可见其顶端细胞发生大量凋亡,免疫荧光检测发现在乳鼠的小肠绒毛顶端及肾脏的肾小球位置均有RV抗原分布。成功构建了猴源轮状病毒SA11株Balb/c乳鼠动物模型,研究发现轮状病毒除感染乳鼠小肠绒毛导致其病变破损发生腹泻外,还可感染肾脏等肠道外器官,引起肠道外感染。
In order to study the pathogenesis of rotavirus(RV),Balb/c neonatal mices were developed for diarrhea model using rotavirus strain SA11.These data provide theoretical and experimental basis for vaccine protection evaluation.The 7-day-old Balb/c mices were intragastrically administrated with RV SA11 strain in different doses.The diarrhea status of these mice were observed at different time points.These mices were euthanized and dissected.Heart,liver,spleen,lung,small intestine and kidney and other tissues of euthanized mices were collected and detected by immunofluorescence for Rotavirus distribution.The collected tissues were fixed in formalin and used in HE staining and IHC staining for the detection of pathological changes of jejunum and the apoptosis of the cells.Obvious diarrhea symptoms were observed at 24 h post inoculation with Rotavirus strain SA11.Pathological anatomy results showed that the intestines of these model mices were congested and flattened after 72 h infection.Compared to the controls,the results of intestine HE staining showed that the pathological changes in model mice were obvious,including vacuolar degeneration,edema and congestion of intestinal wall,integrity destruction of enteric epithelium and cell apoptosis,after intragastrically administered with SA11 strain.Rotavirus distribution in intestine and kidneys was also detected by immunofluorescence assay.In this study,the rotavirus diarrhea models of Balb/c neonatal mice were developed successfully with RV SA11 strain.In addition to small intestine distribution,we found that Rotavirus distribution could also be detected in kidney of Balb/c neonatal mice after infection.
作者
乔洪图
周艳
尹娜
陈林林
刘洋
苏婷
孙茂盛
李鸿钧
QIAO Hong-tu;ZHOU Yan;YIN Na;CHEN Lin-lin;LIU Yang;SU Ting;SUN Mao-sheng;LI Hong-jun(Institute of Medical Biology,Chinese Academy of Medical Science,Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research&Development on Severe Infectious Disease,Kunming 650118,China)
出处
《生物学杂志》
CAS
CSCD
北大核心
2018年第6期22-26,共5页
Journal of Biology
基金
中国医学科学院医学与健康科技创新工程协同创新团队项目(2016-I2M-3-026)
云南省应用基础研究计划项目(2016FB034)
云南省科技计划项目(2014BC008)。
作者简介
乔洪图,硕士,研究方向为生物化学与分子生物学,E-mail:18388240662@163.com;通信作者:李鸿钧,研究员,长期从事病毒学及分子生物学相关研究,E-mail:lihj6912@hotmail.com。
