CAJ | 학술논문

目的构建干扰素(IFN)-α2b基因重组双歧杆菌(pBAD-SPIFN转化双歧杆菌),观察重组菌是否对小鼠柯萨奇B3病毒(CVB3)诱导的小鼠心肌炎具有治疗作用.方法体外扩增前期研究中构建的IFN-α2b重组双岐杆菌.选取BLAB/c小鼠40只,腹腔内注射CVB3感染剂量,2周后形成病毒性心肌炎,将感染的小鼠随机分成IFN、BIFN、B、生理盐水组予以干预:IFN组给予肌肉注射干扰素,BIFN组小鼠予灌饲干扰素-α2b重组双岐杆菌;B组给予灌饲pBAD-gIIIA转化双歧杆菌;生理盐水组给予肌肉注射生理盐水.所有小鼠均在治疗14d后取心脏标本观察心肌组织病理变化,检测心肌病毒滴度,实时荧光定量聚合酶链反应(FQ-PCR)分析Th1细胞因子和IFN-α诱导黏液病毒抗性蛋白1(Mx1)基因转录水平.结果 CVB3诱导的心肌炎小鼠干预治疗2周后,BIFN组小鼠心肌炎症程度(0.16±0.10)较B组及生理盐水组明显减轻(P＜0.01);BIFN组心肌的病毒滴度水平(3.03 ±0.02)较B组及生理盐水组显著下降(P ＜0.01);BIFN组Th1细胞因子和IFN-α诱导基因Mx1基因转录水平(分别为1.48±0.08、3.56±0.02、2.13 ±0.01)较B组及生理盐水组显著升高(P＜0.01).结论灌饲给药IFN-α2b重组双岐杆菌对CVB3病毒诱导的小鼠心肌炎具有较好疗效.
목적 구건간우소(IFN)-α2b기인중조쌍기간균(pBAD-SPIFN전화쌍기간균),관찰중조균시부대소서가살기B3병독(CVB3)유도적소서심기염구유치료작용.방법 체외확증전기연구중구건적IFN-α2b중조쌍기간균.선취BLAB/c소서40지,복강내주사CVB3감염제량,2주후형성병독성심기염,장감염적소서수궤분성IFN、BIFN、B、생리염수조여이간예:IFN조급여기육주사간우소,BIFN조소서여관사간우소-α2b중조쌍기간균;B조급여관사pBAD-gIIIA전화쌍기간균;생리염수조급여기육주사생리염수.소유소서균재치료14d후취심장표본관찰심기조직병리변화,검측심기병독적도,실시형광정량취합매련반응(FQ-PCR)분석Th1세포인자화IFN-α유도점액병독항성단백1(Mx1)기인전록수평.결과 CVB3유도적심기염소서간예치료2주후,BIFN조소서심기염증정도(0.16±0.10)교B조급생리염수조명현감경(P＜0.01);BIFN조심기적병독적도수평(3.03 ±0.02)교B조급생리염수조현저하강(P ＜0.01);BIFN조Th1세포인자화IFN-α유도기인Mx1기인전록수평(분별위1.48±0.08、3.56±0.02、2.13 ±0.01)교B조급생리염수조현저승고(P＜0.01).결론 관사급약IFN-α2b중조쌍기간균대CVB3병독유도적소서심기염구유교호료효.
Objective To investigate a novel oral delivery system for interferon-alpha2b using genetically engineered Bifidobacterium longum as the carrier and further evaluate the efficacy of interferon (IFN)-α2b-expressed B.longum on the coxsackie B3 virus (CVB3)-induced myocarditis in mice.Methods IFN-α2b recombinant B.longum was amplified in vivo and subsequently,BLAB/c mice were inoculated intraperitoneally with infectious dose of CVB3 for two weeks to produce the models of CVB3-induced myocarditis.Then,the murine models were divided into four groups.The "BIFN group" was orally administered with IFN-α2b-transformed B.longum for two weeks respectively after the inoculation of the virus."Plasmid control group" was orally administered with plasmid control-transformed B.longum." Saline group" was administered intraperitoneally with sterile phosphate buffer (PBS)."IFN group" was intramuscullarly injected with IFN-α2b.All animals were killed on the day 14 post-treatment,and the murine hearts were dissected aseptically for hematoxylin-eosin (HE) staining,viral titration and RNA extraction for interferon-induced Mx1 gene transcriptional quantification.Results The data indicated that oral administration of IFN-α2b-expressed B.longum for two weeks after CVB3 infection in the BLAB/c mice could alleviate the severity of virus-induced myocarditis (0.16 ± 0.10,P ＜ 0.01),markedly reduce the virus titers in the heart (3.03 ±0.02),and induce IFN-γ,tumor necrosis factor (TNF)-α and Mx1 gene transcription (1.48 ± 0.08,3.56 ± 0.02,2.13 ± 0.01,respectively) in the heart compared with the controls (P ＜ 0.01).Conclusion Oral administration of IFN-α2b recombinant B.longum may play a therapeutic role in the treatment of CVB3-induced myocarditis in the mice.