β溶血性链球菌M蛋白疫苗对RF/RHD小鼠心脏的保护作用  

Heart protection of M protein vaccine of beta-hemolytic streptococci in RF/RHD mice

作　　者：丁月霞[1] 张铭华[1] 何梅香[1] 张林[1] 陈益民[1] DING Yue - xia, ZHANG Ming - hua, HE Mei - xiang, ZHANG Lin, CHEN Yi - min.(Department of Cardiology, the Fifth Affiliated Hospi- tal, Guangzhou Medical University, Guangzhou 510700, Guangdong, Chin)

机构地区：[1]广州医科大学附属第五医院心血管内科,广东广州510700

出　　处：《广东医学》2018年第18期2727-2731,共5页Guangdong Medical Journal

基　　金：广东省科技计划项目(编号:2014A020212743)

摘　　要：目的β溶血性链球菌(BHS) M蛋白疫苗对风湿热/风湿性心脏病(RF/RHD) BALB/c小鼠心脏的保护作用。方法通过Fmoc固相多肽合成法和反相高效液相色谱法获得15型BHS M蛋白信号肽后35个氨基酸残基和J14肽串联的多肽作为疫苗; BALB/c小鼠随机分成4组:正常组给予PBS和佐剂等量混匀,BHS感染组给予灭活的BHS和佐剂等量混匀; M蛋白疫苗组和预接种M蛋白疫苗组开始均给予M蛋白疫苗与等量佐剂直接多点皮下注射(初次免疫均与弗氏完全佐剂等量混匀、每只小鼠0. 2 m L;之后均和不完全弗氏佐剂等量混匀、每只小鼠0. 2 m L,佐剂使用下同)。最终获取心脏标本通过HE染色,在光镜下观察心肌组织是否形成风湿热小体;并在透射电镜下观察心肌细胞的形态学改变;最后各组眼球采血以测定氨基末端B型利钠肽前体(NT-pro-BNP)、超敏C反应蛋白(hs-CRP)和肌钙蛋白I(c Tn I)水平,进行组间比较。结果正常组和M蛋白疫苗组BALB/c小鼠心脏大体结构正常,光镜下未见异常,BHS感染组心脏标本有化脓表现,光镜下观察到心肌坏死,未观察到风湿热小体。电镜下观察到正常组和M蛋白疫苗组小鼠心肌细胞结构正常、线粒体饱满、轮廓清晰,基质无水肿; BHS感染组心肌细胞线粒体肿胀明显、心肌纤维断裂、水肿,基质肿胀;无论光镜还是电镜下预接种M蛋白疫苗组的结果与正常组心肌细胞形态学一致。与正常组比较,BHS感染组的外周血NT-pro-BNP、c Tn I和hs-CRP水平明显升高(P <0. 05);与BHS感染组比较,预接种M蛋白疫苗组外周血NT-pro-BNP、c Tn I和hs-CRP均较BHS感染组明显降低(P <0. 05),而正常组与M蛋白疫苗组比较,差异无统计学意义(P> 0. 05)。结论 M蛋白疫苗预接种RF/RHD小鼠后对BHS致心脏损伤作用有明显减轻效果,并显著降低外周血NT-pro-BNP、c Tn I和hs-CRP水平。Objective To investigate the protective effects of beta-hemolytic streptococci（ BHS） M protein vaccine on hearts of BALB/c mice with rheumatic fever/rheumatic heart diseases（ RF/RHD）. Methods M protein vaccine was conducted using 35 amino acids and J14 serial polypeptide. The BALB/c mice were randomly divided into four groups,normal group,M protein vaccine group,BHS infection group and pre-vaccination group（ M protein vaccine was offered and with multiple BHS infection）. Rheumatoid bodies in myocardial tissues were observed by light microscope,and morphological changes of myocardial cells were also observed by transmission electron microscope. Blood samples were collected from eyeballs for assessment of NT-pro-BNP,cTnI and hs-CRP. Results The general cardiac structures of BALB/c mice in normal group and M protein vaccine group were normal. There was no abnormality observed under light microscope. There were fester representation and myocardial necrosis in cardiac specimen observed in BHS infection group,but no rheumatoid body was observed under light microscope. It＇s observed through electron microscope that myocardial structures in normal group and M protein vaccine group were normal; with voluptuous mitochondria with clear boundaries,and no edema in matrix. Mitochondrial swelling in myocardial cells,ruptured myocardial fiber with edema and swelling matrix were observed in BHS infection group. Morphological results in pre-vaccination group were consistent with those in normal group both under light microscope and electron microscope. Compared with normal group and M protein vaccine group,NT-pro-BNP,cTnI and hs-CRP levels of peripheral blood in BHS infection group were significantly increased（ P〈 0. 05）. Compared with BHS infection group,NT-pro-BNP,cTnI and hs-CRP level of peripheral blood in pre-vaccination group were all significantly reduced（ P〈 0. 05）; with no significant difference when comparing tonormal group and M protein vaccine group（ P 〉0. 05）. Conclusion The BHS M prote

关 键 词：Β溶血性链球菌 M蛋白多肽疫苗 风湿热/风湿性心脏瓣膜病 

分 类 号：R392[医药卫生—免疫学]