柯萨奇病毒 B3诱导的病毒性心肌炎 C57BL／6J小鼠模型的建立

摘要

Aim:To establish a C57BL/6J mouse model of myocarditis induced by coxsackievirus B 3(CVB3).Meth-ods:A total of 50 male mice with 4-week-age or 6-week-age were divided into two groups:the control(n=15) were injec-ted via intraperiton with PBS, and the model group(n=35) were injected via intraperiton with 1 ×105 PFU CVB3.After 14 d, mice hearts were harvested for HE staining and CD 3,CD45 immunohistochemistry staining ,the serum levels of IL-1β,TNF-αwere assayed by ELISA , the expressions of p-AKT,p-IkBαand p-GSK3βwere detected by Western blot .Re-sults:4-week-age model group came out with a higher mortality (19/35,54.3%), HE staining showed that a great deal of inflammatory cells concentrated in the adjacent area around the myocardiocytes ,which was consistent with the finding that many CD3,CD45 positive macrophages gathered in the necrosis area ,TNF-αand IL-6 level were increased(P<0.05),p-AKT,p-IκBαand p-GSK3βprotein expression increased (P<0.05).While the 6-week-age model group showed a lower mortality(3/35,8.6%),and the immune response to CVB3 infection was milder than 4-week-age model group(P<0.05). Conclusion:A C57BL/6J mouse model of myocarditis induced by CVB 3 is successfully established ,and 4-week-age mice are recommended for this model .%目的：建立由柯萨奇病毒B3（CVB3）Nancy株诱导的病毒性心肌炎（VMC） C57BL／6J小鼠模型。方法：4周龄、6周龄C57BL／6J小鼠各50只，每个年龄组随机选取15只作为对照（腹腔注射PBS），剩余35只腹腔接种含CVB3 Nancy株的病毒悬液（1×105 PFU／只），于感染后第14天取心脏，行HE染色和CD3、CD45免疫组化染色，进行病理学评价，检测血清白细胞介素-1β（ IL-1β）、肿瘤坏死因子-α（ TNF-α）水平，并做心肌组织中病毒滴度检测，采用Western blot法检测磷酸化AKT、IkBα、GSK3β的表达。结果：4周龄小鼠造模过程中有19只（54．3％）死亡；与对照小鼠比较，模型小鼠心肌呈现弥漫性炎性细胞浸润和片状心肌细胞变性坏死， CD3、CD45阳性细胞弥漫浸润于心肌间质，血清IL-1β、TNF-α水平明显升高（P＜0．05），心肌组织中磷酸化AKT、IkBα、GSK3β表达增高（P＜0．05）。6周龄小鼠造模过程中有3只（8．6％）死亡，模型小鼠心肌组织中炎症反应轻于4周龄模型小鼠（P＜0．05）。结论：成功建立了VMC的C57BL／6J小鼠模型，4周龄C57BL／6J小鼠更适用于模型的建立。