摘要
目的探讨IL-13基因质粒水平基因转移(pIL-13 HGT)缓解柯萨奇病毒B3型(CVB3)诱导BALB/c小鼠急性病毒性心肌炎的机制。方法将BALB/c小鼠分为十二组,每组6～8只。A组为对照;其他组小鼠按照每只10~3半数组织培养感染剂量(TCID_(50))/100μl PBS的病毒量腹腔注射,建立CVB3诱导的急性病毒性心肌炎小鼠模型。B组采用脂多糖(LPS)+IFN-γ治疗;C组采用LPS+IFN-γ+IL-13治疗;D组单用IL-13治疗;E组没有经载体HGT处理;F组经载体HGT空质粒处理;G组经pIL-13HGT治疗;H组不做任何治疗作为阳性对照;四个采用不同IL-13用量(0、0.2、2和20ng/ml)治疗组。采用流式细胞术分析F4/80^+CD206^+巨噬细胞比例和CD206的表达,ELISA法检测各细胞因子表达水平。结果随着IL-13浓度的升高,F4/80^+骨髓巨噬细胞(BMDM)的CD206和IL-10的表达也增加(P<0.05)。B组BMDM的TNF-α、IL-12表达高于A、C和D组(P<0.05),而IL-10表达低于C、D组(P<0.05)。B组的IL-6表达高于A、D组(P<0.05)。G组心脏浸润细胞和脾细胞中F4/80^+CD206^+巨噬细胞比例和F4/80^+巨噬细胞荧光强度高于E、F组(P<0.05)。与F组相比,G组心脏组织和脾脏组织中TNF-α表达降低(P<0.05),而IL-10表达增加(P<0.05)。E组心脏浸润细胞中CD3^+CD4^+T细胞比例高于F、G组(P<0.05)。G组心脏浸润细胞中F4/80^+巨噬细胞比例高于E、F组(P<0.05)。G组血清中IFN-γ表达低于E、F组,而IL-4表达高于E、F和H组(P<0.05)。E、F组血清中Th1细胞的比例高于G、H组(P<0.05)。结论 IL-13在CVB3诱导的病毒性心肌炎中起保护性作用,而M2型巨噬细胞和Th2应答可能均参与了这一过程。
Objective To investigate the mechanism of IL-13 gene plasmid horizontal gene transfer(pIL-13 HGT)in relieving acute viral myocarditis induced by CVB3 in BALB/c mice.Methods BALB/c mice were assigned into 12 groups with 6-8 mice each.Group A was taken as the control.The CVB3-induced acute viral myocarditis mouse models were established in the other 11 groups,which was performed by intraperitoneally injecting virus in a dose of 103 50% tissue culture infecting dose(TCID50)/100μl PBS.Group B was treated with lipopolysaccharide(LPS)+IFN-γ,group C was treated with LPS+IFN-γ+IL-13,group D was treated with IL-13,group E was treated without vector HGT,group F was treated with vector HGT empty plasmid(vector HGT empty plasmid group),group G was treated with pIL-13 HGT,group H was not treated with any drug as positive control,and four groups were treated with different amounts of IL-13(0,0.2,2,20 ng/ml).The ratio of F4/80+CD206+macrophages and the expression of CD206 were analyzed by flow cytometry.The expression of cytokines was detected by ELISA.Results With the increase of IL-13 concentration,the expressions of CD206 and IL-10 were increased in F4/80+BMDM(P<0.05).The expressions of TNF-αand IL-12 in group B were higher than those in groups of A,C and D(P<0.05),while IL-10 expression was lower than that in groups of C and D(P<0.05).The expression of IL-6 in group B was higher than that in groups of A and D(P<0.05).The proportion of F4/80+CD206+macrophages and the fluorescence intensity of F4/80+macrophages of heart infiltrates cells and spleen cells in group G were higher than those in groups of E and F(P<0.05).Compared with group F,the expression of TNF-αin heart and spleen tissues of group G was decreased(P<0.05),while the expression of IL-10 was increased(P<0.05).The proportion of CD3+CD4+T cells of heart infiltrates cells in group E was higher than that in groups of F and G(P<0.05).The proportion of F4/80+macrophages of heart infiltrates cells in group G was higher than that in groups of E and F(<0.05).Serum level of IFN-γexpression in group G was lower than that in groups of E and F,and the IL-4 expression was higher than that in groups of E,F and H(P<0.05).Serum proportion of Th1 cells in groups of E and F was higher than that in groups of G and H(P<0.05).Conclusion IL-13 plays a protective role in CVB3-induced viral myocarditis and both M2 macrophages and Th2 responses may be involved in this process.
作者
韩秀江
高晟
徐新建
HAN Xiujiang;GAO Sheng;XU Xinjian(Department of Cardiology,Tianjin Hospital of ITCWM Nankai Hospital,Tianjin 300100,CHINA)
出处
《江苏医药》
CAS
2018年第11期1231-1235,共5页
Jiangsu Medical Journal
基金
天津市卫生部立项项目(2015029)
