柯萨奇病毒性心肌炎研究进展

柯萨奇病毒性心肌炎研究进展上海第二医科大学微生物教研室解立颖综述刘晶星，陆德源审校小核糖核酸科的肠道病毒如何萨奇病毒Ｂ组（ＣｏｘｓａｃｋｉｅｖｉｒｕｓＢ，ＣＢ）是病毒性心肌炎的主要病原。肠道病毒引起的心肌炎可以是急性的或慢性的，病人可以完全康复、慢性...     

柯萨奇病毒致病毒性心肌炎分子生物学机理

柯萨奇B组病毒是引起病毒性心肌炎的主要病原之一，有25％VMC患者的确切病因是由CVB感染所致。根据其对新生乳鼠的致病特点，CVB可分为六个血清型，其中CVB3有很强的噬心肌性，在病变心肌组织中发现的CVB3病毒RNA序列已经证实这一点。通过研究CVB3病毒结构蛋白与疾病发病机理之间的关系，以及重组CVB3嵌合病毒与心血管疾病的相关位点，证实CVB3的5’端非编码区对病毒毒力的重要作用；同时证实除诱导心肌炎的病毒可以直接破坏感染细胞外，宿主的自发性免疫反应则使心肌组织进一步损伤，由于心肌纤维化、心肌细胞肥大以及心肌结构重塑，最终导致约15％的VMC患者可逐渐发展成为扩张型心肌病。     

柯萨奇-腺病毒受体与病毒性心肌炎

柯萨奇 腺病毒受体是一种普通的存在于心肌细胞表面的病毒受体 ,作为柯萨奇病毒和腺病毒的共同受体 ,在它们感染心肌细胞的过程中起着关键性的作用。     

柯萨奇B4病毒感染与克山病及其病区病毒性心肌炎

目的 探讨柯萨奇B4病毒与克山病及其病区病毒性心肌炎的关系。方法 采用寡核苷酸探针的原位核酸杂交方法,结合病理学观察,对比研究克山病与病毒性心肌炎的异同。结果 克山病与病毒性心肌炎各自具有特征性的病理学改变;发病区88．89％的病毒性心肌炎出现阳性杂交信号,而且几乎每例的所有心肌细胞均见阳性杂交信号;95．24％的亚急型克山病出现阳性杂交信号,且半数以上的标本几乎所有的心肌细胞内出现阳性杂交信号;阳性信号在病毒性心肌炎和亚急型克山病的心肌细胞内的分布及阳性信号颗粒的形态并无明显区别。结论 柯萨奇B4病毒与克山病及其病区病毒性心肌炎密切相关,其不同的病变特征可能是由于组成病因网的诸因素发生变动所致。     

柯萨奇B_4病毒感染与克山病及其病区病毒性心肌炎

目的　探讨柯萨奇 B4病毒与克山病及其病区病毒性心肌炎的关系。方法　采用寡核苷酸探针的原位核酸杂交方法 ,结合病理学观察 ,对比研究克山病与病毒性心肌炎的异同。结果　克山病与病毒性心肌炎各自具有特征性的病理学改变 ;发病区 88.89%的病毒性心肌炎出现阳性杂交信号 ,而且几乎每例的所有心肌细胞均见阳性杂交信号 ;95 .2 4%的亚急型克山病出现阳性杂交信号 ,且半数以上的标本几乎所有的心肌细胞内出现阳性杂交信号 ;阳性信号在病毒性心肌炎和亚急型克山病的心肌细胞内的分布及阳性信号颗粒的形态并无明显区别。结论　柯萨奇 B4病毒与克山病及其病区病毒性心肌炎密切相关 ,其不同的病变特征可能是由于组成病因网的诸因素发生变动所致     

细胞因子和病毒性心肌炎的关系

本文着重阐述了细胞因子在病毒性心肌炎发生，发展中的作用。     

中医药防治柯萨奇B3病毒性心肌炎研究概述

病毒性心肌炎 (VMC)是以柯萨奇B3 病毒性感染为主的临床常见心脏疾患。近年来 ,国内发病率有所增高。VMC与扩张型心肌病 (DCM )之间的关系也日益为人们所重视。许多资料表明[1] ,通过临床随访 ,确诊为VMC的 10 %～ 5 0 %最终演变为DCM。由于DCM预后不良 ,病人除进行高度心脏移植手术别无其他特效治疗手段 ,故VMC的防治引起临床重视。自 2 0世纪 80年代以来 ,国内运用中医药或中西医结合对防治柯萨奇B3 病毒性心肌炎 (CVB3 -VMC)进行了实验和临床研究 ,大量研究证明 ,中医药在防治CVB3 -VMC上具有一定的优势 ,本文就其主要技术…     

黄芪甲甙对小鼠柯萨奇B3病毒性心肌炎的抗氧化作用

目的观察黄芪甲甙对小鼠柯萨奇B3病毒性心肌炎的抗氧化作用。方法Balb/c小鼠50只随机分5组(每组10只),空白对照组腹腔无菌注射不含病毒的Eagle’s培养基0.1 mL,在腹腔注射病毒培养基30 min后,以生理盐水0.1 mL灌胃,共7d;病毒性心肌炎对照组小鼠每只腹腔注射0.1 mL内含1×102TCID50柯萨奇B3病毒的Eagle’s培养基,在腹腔注射含病毒的Eagle’s培养基30 min后,以生理盐水0.1 mL灌胃,共7 d;黄芪甲甙低、中、高剂量干预组在腹腔注射病毒30 min后,用黄芪甲甙剂量分别为0.07、0.2、0.6 g/(kg.d)0.1 mL灌胃,共7 d。观察小鼠生存数;心肌病变积分;心肌谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力及逆转录聚合酶链反应检测铜锌超氧化物歧化酶-mRNA水平表达。结果(1)与空白对照组比较,病毒性心肌炎对照组小鼠的生存数明显下降,心肌病变积分明显升高(3.23±0.83,P<0.01);与病毒性心肌炎对照组比较,黄芪甲甙高剂量干预组可明显提高小鼠的生存数(P<0.01),心肌病变积分明显下降(1.86±0.59比3.23±0.83,P<0.01);(2)与病毒性心肌炎对照组比较,黄芪甲甙高剂量干预组心肌组织谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力呈增高趋势,心肌组织谷胱甘肽过氧化物酶(39.73±8.02比9.99±2.71,P<0.05)、过氧化氢酶(6.35±2.33比1.36±0.87,P<0.05)、超氧化物歧化酶(65.71±4.93比40.15±6.03,P<0.01)活力明显增强,铜—锌超氧化物歧化酶-mRNA水平表达明显增高(0.43±0.11比0.32±0.01,P<0.01)。但黄芪甲甙低、中剂量干预组,虽然也在一定程度使心肌组织谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活力及逆转录聚合酶链反应检测铜—锌超氧化物歧化酶-mRNA表达水平增加,有一定的量效关系,但与病毒性心肌炎对照组比较无统计学意义(P>0.05)。结论黄芪甲甙通过增加心肌抗氧化酶活力,对小鼠柯萨奇B3病毒性心肌炎具有明显的保护作用。     

111例成人急性柯萨奇病毒B心肌炎后室性早搏的长期随访

119例经血清学与临床诊断为急性柯萨奇病毒B心肌炎的病人,长期随访平均3.5年(4—98月),随访率93.3％。急性期后82％的患者有室性早搏长期间断发作。随访期间无一例发生昏厥、近乎昏厥、猝死或心电图证实的持久性室性心动过速发作,亦无扩张型心肌病的临床、X线或超声心动图表现。     

信号转导和转录激活子3与病毒性心肌炎的关系

目的:探讨信号转导和转录激活子3(STAT3)在病毒性心肌炎(VMC)发病机制中的作用.方法:以柯萨奇病毒B3感染balb/c小鼠,建立急慢性VMC的系列动物模型;经组织病理学方法证实模型后,采用免疫印迹法(Western blot)和免疫组化检测心肌组织细胞质和细胞核中STAT3和磷酸化STAT3(p-STAT3)蛋白.结果:Western-blot和免疫组化法显示柯萨奇病毒B3感染小鼠心肌细胞核STAT3和p-STAT3蛋白含量较对照组明显增高(P＜0.05).细胞质STAT3和p-STAT3蛋白含量与对照组比较差异无统计学意义(P＞0.05).结论:VMC时心肌细胞核STAT3和p-STAT3蛋白明显增高,与其发病密切相关.     

Effects of granulocyte colony-stimulating factor upon coxsackievirus B3 myocarditis in mice

Granulocyte colony-stimulating factor is a potent activatorof mature granulocytes, and subsequently enhances superoxiderelease. The purpose of this study was to investigate the effectsof granulocyte colony-stimulating factor upon murine coxsackievirusB3 myocarditis in relation to free radical-mediated cardiacdamage. Two-week-old, male, C₃H/He mice were inoculated intraperitoneallywith coxsackievirus B3. Gramulocyte colony-stimulating factor20 µg.kg^–1. day^–1, polyethylene glycol-conjugatedsuperoxide dismutase (an enzyme catalyzing the conversion ofO₂^- to H₂O₂) 1 x 10³ U.kg^–1 day^–1 and granulocytecolony-stimulating factor 20 µg. kg^–1 day^–1,plus polyethylene glycol-conjugated superoxide dismutase 1 x10³ U.kg^–1. day^–1, were injected subcutaneouslydaily on days 0 to 14. Treated groups were compared to the infected,untreated group. The survival rate in the polyethylene glycol-conjugatedsuperoxide dismutase group was higher than that of the untreatedgroup on day 14, but on day 7, cardiac pathology was not significantlydifferent among the four groups. On day 14, the scores of cellularinfiltration, myocardial necrosis and calc were lower in thepolyethylene glycol-conjugated superoxide disnuitase group andin the granulocyte colony-stimulating factor plus polyethyleneglycol-conjugated superoxide dismutase group than in the untreatedgroup. The myocardial virus titres on days 7 and 14 did notd sign y among the four groups. The number of total white bloodcell and neutrophil counts were signifIcantly greater in thegranulo cyte colony-stimulating factor group than in the untreatedgroup on day 7. Taken altogether with the previous reports andpresent evidence that the administration of granulocyte colony-stimulatingfactor did not exacerbate coxsackievirus B3 myocarditis, itmay be that oxygen-free radicals appeared to be derived notfrom leukocytes but from other components in this experimentalmodel of myocarditis, whereas the myocardium was inflamed withleukocytes.     

Effects of granulocyte colony-stimulating factor upon coxsackievirus B3 myocarditis in mice

Granulocyte colony-stimulating factor is a potent activatorof mature granulocytes, and subsequently enhances superoxiderelease. The purpose of this study was to investigate the effectsof granulocyte colony-stimulating factor upon murine coxsackievirusB3 myocarditis in relation to free radical-mediated cardiacdamage. Two-week-old, male, C₃H/He mice were inoculated intraperitoneallywith coxsackievirus B3. Gramulocyte colony-stimulating factor20 µg.kg^–1. day^–1, polyethylene glycol-conjugatedsuperoxide dismutase (an enzyme catalyzing the conversion ofO₂^- to H₂O₂) 1 x 10³ U.kg^–1 day^–1 and granulocytecolony-stimulating factor 20 µg. kg^–1 day^–1,plus polyethylene glycol-conjugated superoxide dismutase 1 x10³ U.kg^–1. day^–1, were injected subcutaneouslydaily on days 0 to 14. Treated groups were compared to the infected,untreated group. The survival rate in the polyethylene glycol-conjugatedsuperoxide dismutase group was higher than that of the untreatedgroup on day 14, but on day 7, cardiac pathology was not significantlydifferent among the four groups. On day 14, the scores of cellularinfiltration, myocardial necrosis and calc were lower in thepolyethylene glycol-conjugated superoxide disnuitase group andin the granulocyte colony-stimulating factor plus polyethyleneglycol-conjugated superoxide dismutase group than in the untreatedgroup. The myocardial virus titres on days 7 and 14 did notd sign y among the four groups. The number of total white bloodcell and neutrophil counts were signifIcantly greater in thegranulo cyte colony-stimulating factor group than in the untreatedgroup on day 7. Taken altogether with the previous reports andpresent evidence that the administration of granulocyte colony-stimulatingfactor did not exacerbate coxsackievirus B3 myocarditis, itmay be that oxygen-free radicals appeared to be derived notfrom leukocytes but from other components in this experimentalmodel of myocarditis, whereas the myocardium was inflamed withleukocytes.     

Effect of interleukin-15 on the course of myocarditis in Coxsackievirus B3-infected BALB/c mice

OBJECTIVE:

Cytokines have an important role in both the initiation and perpetuation of viral myocarditis. Because a causative therapy of myocarditis is not yet well established and immunomodulation is a promising approach, the influence of interleukin (IL)-15, a proinflammatory cytokine, on the course of experimental myocarditis in Coxsackievirus B3 (CVB3)-infected mice was examined.

METHODS:

Hearts from CVB3-infected (n=14), sham-infected (n=14) and CVB3-infected BALB/c mice treated with IL-15 (n=6) or a competitive IL-15 fusion protein (n=6) were analyzed for hemodynamic function, cellular infiltrates and myocardial collagen content.

RESULTS:

Induction of myocarditis was associated with significant loss of body and heart weight, decreased left ventricular function, and increased collagen content and cellular infiltrates in the myocardium. Treatment of infected animals with IL-15 resulted in normalization of body and heart weight, and significantly improved systolic and diastolic left ventricular function, comparable with that of uninfected animals. This was paralleled by a significant reduction of myocardial collagen content to levels observed in animals without disease and by markedly reduced cellular infiltration of lymphocytes and macrophages in the myocardium. Inhibition of intrinsic IL-15 with IL-15 fusion protein tended to aggravate the disease.

CONCLUSIONS:

Treatment with IL-15 has a positive effect on CVB3-induced murine myocarditis and seems to be a promising approach to modifying clinical course, hemodynamics and histopathology of virus-induced myocarditis. Further studies are needed to identify the underlying mechanisms.     

Protection against murine coxsackievirus B3 myocarditis by T cell vaccination

T cell vaccination regulates autoimmunity by the modification of helper and suppressor T cells. The present study was performed to examine whether T cell vaccination can prevent viral myocarditis in vivo. We used coxsackievirus B3 myocarditis in mice as an animal model with the analysis of lymphokine-activated killer cell activity. Vaccination of the mice with T lymphocytes significantly prolonged survival and improved cardiac histology of murine myocarditis. The effects of T cell vaccination were most evident when T cells sensitized with the same virus were used. Vaccination of the mice with T cells from other strains of mice showed lesser protective effects. Clearance of myocardial virus was not affected by this treatment. The efficacy of T cell vaccination was confirmed in vitro by the decrease of the lymphokine-activated killer cell activity against EL-4 tumor cells and cultured myocytes. T cell vaccination of mice prolonged survival and improved myocardial lesions of animals inoculated with coxsackievirus B3.     

Amyocarditic coxsackievirus B3 causes myocarditis in immunocompromised mice

In the present study, we investigated the role of the immune status of the host in the pathogenesis and development of coxsackievirus B3 myocarditis. We compared the disease expression in myocarditic coxsackievirus B3 (CB3M)-infected BALB/c wild-type mice and severe combined immunodeficient (SCID) mice and in amyocarditic coxsackievirus B3 (CB3O)-infected BALB/c wild-type mice untreated or treated with immunosuppressive agents and SCID mice. There were no differences in viral growth in vitro between CB3M and CB3O. Severe myocarditis developed in CB3M-infected wild-type and SCID mice, CB3O-infected SCID mice and CB3O-infected wild-type mice with total immunosuppression. However, myocarditis was not induced in CB3O-infected wild-type mice untreated and treated with partial immunosuppression. There were no changes in myocardial virus titres among these groups of mice. In addition, myocarditis was induced in CB3O-infected wild-type mice treated with Thy 1.2 (pan T) or Lyt 2 (CD8) antibody but not in those mice treated with L3T4 (CD4) antibody. Thus, the CB3O variant did not induce myocarditis in wild-type mice associated with the induction of the CD8⁺ lymphocyte subset but was shown to have the genetic capability to induce myocarditis if the host was in an almost total immunosuppressive or CD8-depleted state. The results suggest that induction of myocarditis by the amyocarditic strain of coxsackievirus B3 may occur and partially depends on the immune status of the host, and that myocarditis is due in part to an immunopathogenic mechanism.     

外源性细胞因子对柯萨奇病毒性心肌炎小鼠脾Th17细胞的影响

目的:观察转化生长因子(TGF)-β、白细胞介素(IL)-6和IL-23对柯萨奇病毒性心肌炎(VMC)小鼠脾中Th17细胞分化增殖的影响。方法:以6周龄雄性Balb/c小鼠腹腔注射柯萨奇病毒液建立VMC模型,1周后磁珠分离脾脏CD4+T淋巴细胞,分成对照组、TGF-β+IL-6组和TGF-β+IL-6+IL-23组进行体外培养。5d后应用流式细胞术测定刺激后Th17细胞的百分比;逆转录-聚合酶链式反应测定细胞中IL-17mRNA的表达;ELISA测定细胞培养上清液IL-17的水平。结果:与对照组比较,TGF-β+IL-6组Th17细胞数稍增加[(1.64±0.43)%∶(1.96±0.35)%],IL-17mRNA表达水平及细胞培养上清液IL-17浓度稍增加,但差异无统计学意义(P>0.05)。而TGF-β+IL-6+IL-23组Th17细胞数显著增加(4.08±0.77)%、IL-17mRNA表达水平及细胞培养上清液IL-17浓度均明显高于前2组,差异有统计学意义(P<0.05)。结论:外源性TGF-β联合IL-6对VMC小鼠脾中已分化的Th17细胞无增殖作用,而添加外源性IL-23则能显著增加Th17细胞数及其效应分子IL-17的表达。     

应用嗜鼠心肌CoxsackieB3病毒反复感染致BALB／c小鼠心肌损伤

应用嗜鼠心肌ＣｏｘｓａｃｋｉｅＢ３病毒（ＣＶＢ３ｍ）给ＢＡＬＢ／Ｃ小鼠重复腹腔注射，连续３次，间隔３０天，末次注射后１０天处死小鼠。光学显微镜检查发现，小鼠心肌可见不同时期的病灶（坏死灶、瘢痕），心肌损伤检出率达１００％。应用ＴＵＮＥＬ法检测发现，８２．５０％感染组鼠心肌中可见散在心肌细胞凋亡，与对照组（２３．０８％）比较，Ｐ＜０．０１，有极显著性差异。提示ＣＶＢ３ｍ重复感染可致ＢＡＬＢ／Ｃ小鼠慢性心肌损伤，心肌细胞凋亡参与心肌损伤的发生与发展。     

抗柯注射液对柯萨奇B3病毒感染小鼠自然杀伤细胞活性的影响

目的 :检测 BAL B/ c小鼠在感染柯萨奇 (cox) B3病毒后自然杀伤 (NK)细胞活性的动态变化以及抗柯注射液 (RSF)对其影响。方法 :将小鼠分为正常对照组、病毒对照组、10 m g/ kg RSF组、2 0 mg/ kg RSF组和 30m g/ kg RSF组 ,每组 17只 ,分别在病毒感染后第 3天、8天、12天用乳酸脱氢酶释放法 ,检测各组小鼠外周血的NK细胞活性 ,并将心脏行组织病理检查。结果 :病毒感染后第 3天 NK细胞活性显著升高 (6 7.11± 2 .5 2 ) % ,之后进行性下降 ,第 12天时低于正常 [(42 .2 6± 1.6 1) %∶ (47.79± 4.0 3) % ];RSF对 cox B3病毒感染小鼠的 NK细胞活性有双向调节作用。大剂量 (2 0 mg/ kg和 30 mg/ kg) RSF在第 3天时对增高的 NK细胞活性有抑制作用 ,第 12天时可使降低的 NK细胞活性恢复至正常水平 ;小剂量 (10 mg/ kg) RSF在第 3天时对增高的 NK活性无抑制作用 ,但第 12天时也能使降低的 NK活性恢复至正常。病毒感染第 8天时心肌病变最重 ,小剂量 RSF组心肌病变改善较少 ,大剂量 RSF组改善明显。结论 :RSF调节 cox B3病毒感染后机体的 NK细胞活性可能是其防治病毒性心肌炎的重要机制之一。     

硫酸类肝素-3-O-磺基转移酶B1对乙型肝炎病毒复制的影响

目的 研究硫酸类肝素-3-O-磺基转移酶B1(HS3ST381)对HBV复制的影响.方法 以HepG2细胞为阴性对照组,转染2.5μg pCH9-HBV(HBV表达质粒)的HepG2细胞为阳性对照组;转染了2.5μg pCH9-HBV、1.5 μg pcDNA3.1(HS3ST3B1表达质粒载体)和2.0μgpTZU6...