摘要
目的利用不同浓度柯萨奇病毒B组3型(Coxsackievirus B3,CVB3)感染幼龄SD大鼠,探究病毒在大鼠组织分布、免疫应答以及炎症因子的反应,有助于揭示病毒感染的免疫病理,为药物筛选和疗效评价提供实验依据。方法7日龄SD大鼠经腹腔注射不同剂量的CVB3(TCID_(50)为10^(-3.34)/100μL),分别于造模第4、8天采用流式细胞检测全血中淋巴细胞亚群比例(CD4^(+)、CD8^(+)),利用实时荧光定量聚合酶链反应(qRT-PCR)检测心脏、肝、脾、脑、肾、胃肠道组织CVB3载量,采用大鼠ELISA试剂盒检测组织中TNF-α、IFN-γ水平,并采用苏木素-伊红(HE)染色观察组织形态学变化。结果不同剂量的CVB3可引起幼龄大鼠出现不同程度的腹泻,且体质量下降;CVB3主要分布于胃、小肠、大肠组织及粪便,其中在大肠及粪便中载量最高;与正常组比较,原液组(TCID_(50)=10^(-3.34)/100μL)幼龄大鼠全血中CD8^(+)T比例增加和CD4^(+)/CD8^(+)比值减少(P<0.05,P<0.01),大肠组织中TNF-α高表达和IFN-γ低表达(P<0.05,P<0.01),病理组织观察可见大肠组织黏膜下层水肿及炎症细胞浸润,而10-1组、10-2组、10-3组幼龄大鼠全血中淋巴细胞亚群比例、TNF-α、IFN-γ含量及组织形态学均无统计学意义(P>0.05)。结论以不同剂量的CVB3诱导幼龄SD大鼠感染模型,其中原液攻毒CVB3(TCID_(50)=10^(-3.34)/100μL)可引起幼龄大鼠大肠组织病理损伤、病毒大量复制可引起炎性因子水平及免疫细胞失调,提示CVB3病毒对幼龄大鼠具有独特的致病机制,为开发抗CVB3病毒感染药物评价策略提供理论基础。
Objective Using different concentrations of Coxsackievirus B3(CVB3)to infect young SD rats.To investigate the distribution of coxsackievirus B3(CVB3)in rat tissues and the immune response and inflammatory factors,to clarify the immunopathological mechanism of viral infection and provide an experimental basis for drug screening and efficacy evaluation.Methods Young SD rats(7 days old)were injected intraperitoneally with different doses of CVB3(TCID_(50)=10-3.34/100μL)and the proportions of lymphocyte subsets(CD4^(+),CD8^(+))in whole blood at days 4 and 8 were detected by flow cytometry.The CVB3 loads in the heart,liver,spleen,brain,kidney,and gastrointestinal tissues were detected by real-time fluorescence quantitative polymerase chain reaction,TNF-αand IFN-γlevels were detected by enzyme-linked immunosorbent assay,and histomorphologic changes were observed by hematoxylin and eosin staining.Results Different doses of CVB3 caused different degrees of diarrhea and decreased body mass in young rats.CVB3 was mainly distributed in the stomach,small intestine,large intestine,and stools,with the highest load in the large intestine and stools.The stock solution group(TCID_(50)=10-3.34/100μL)increased the proportion of CD8^(+)T cells in the whole blood in young rats and decreased the CD4^(+)/CD8^(+)ratio(P<0.05,P<0.01).Compared with the nomal group high TNF-αand low IFN-γexpression were observed in the large intestine of young rats in the concentrate group(P<0.05,P<0.01),and submucosal edema and inflammatory cell infiltration were observed in the large intestine(cecum and rectum).There were no significant differences in the proportion of lymphocyte subsets,TNF-αand IFN-γlevels,and morphological changes in whole blood of young rats in the group 10-1,10-2,and 10-3(P>0.05).Conclusions Different doses of CVB3 can induce infections in young SD rats.CVB3(TCID_(50)=10-3.34/100μL)causes pathological changes in the large intestine(cecum and rectum)in young rats,and high virus replication can increase levels of inflammatory factors and cause an imbalance of immune cells.CVB3 may have a unique pathogenic mechanism in young rats,providing a theoretical basis for developing evaluation strategies for drugs against CVB3 virus infections.
作者
张卓
刘学武
陈相池
肖洒
杨柳
姜德建
彭冬冬
ZHANG Zhuo;LIU Xuewu;CHEN Xiangchi;XIAO Sa;YANG Liu;JIANG Dejian;PENG Dongdong(Hunan Prima Drug Research Center Co.Ltd,Changsha 410329,China;Hunan Key Laboratory of Pharmacodynamics and Safety Evaluation of New Drugs,Changsha 410329,China)
出处
《中国实验动物学报》
北大核心
2025年第7期1032-1042,共11页
Acta Laboratorium Animalis Scientia Sinica
基金
湖南普瑞玛药物非临床研究创新创业团队(2021)。
关键词
CVB3
幼龄SD大鼠
组织分布
免疫病理
CVB3
young SD rats
tissue distribution
immunopathology Conflicts of Interest:The authors declare no conflict of interest
