The involvement of IL-23/Th17 pathway in mice virus myocarditis inducedby cosakievirus B3

Background The IL-23/ Th17 pathway plays an important role in the development of chronic in? ammatory diseases and autoimmune diseases.However,the role of the IL-23/Th17 axis in the regulation of virus myocarditis is still largely unknown.We aim to determine the role of IL-23/ Th17 axis in virus myocartidis.Methods and Results Balb/c male mice were peritoneally injected with 100TICD50 Coxsackie virus B3 to establish virus myocarditis(VMC), mice injected with PBS peritoneally were taken as the controls. 0,1,2,3,4 and 6 weeks After injection,IL-23,L-17 and RORγt mRNA in the myocardium of VMC were assessed by Semi-quantitative RT-PCR,and IL-23 protein from plasm was evaluated by ELISA.Flow cytometric analysis was used to evaluate the frequencies of Th17 subsets in CD4,To investigated whether the IL-23 is important during IL-23/Th-17 pathway challenge,we isolated CD4 + T cells and cultured with rIL-23 in vitro,and examined the Th17 cells.Results show that,comparing with the controls,IL-23,IL-17 and RORγt mRNA was steadly expressed in the myocardiums of infected mice from 1 week after virus injection.Conclusions IL-23/ Th-17 pathway may therefore play an essential role in VMC. Comparative studies are required to reveal further the roles of these cytokines in the pathogenesis of? these immune-related diseases.

The effect of astragalus membranaceus in combination with taurine on coxsackievirus B3 murine myocarditis

Objective To evaluate the effect of astragalus membranaceus (AM) in combination with taurine on coxasckievirus B3 (CVB3) murine myocarditis. Methods One hundred and sixty mice were infected with CVB3 and equally divided into four groups: normal saline (NS) treated group, AM treated group, taurine treated group and AM combined with taurine treated group. The treatment were carried out from day 1 to 7 after infection. Ten mice were extracted from each group for observing mortality within day 21 postinfection, the rests were killed and the hearts were removed on day 8 (n=15) and 21 (all survival mice), respectively. Electrocardiograph (ECG) of each mouse was analysed before infection and repeated on the day of being killed. CVB3 RNA in murine myocardium was detected by in situ hybridization and myocardial lesions were observed by light microscope. Results Mortality was lower in drug treated groups than that in NS treated group, and it was the lowest in AM combined with taurine treated group, but all the differences were not significant (P>0.05). All drug treatments reduced the incidence of abnormal ECG changes in the acute phase of infection (P<0.05, vs NS treated group), but AM combined with taurine was the most effective in reducing the incidence of abnormal ECG (P<0.05, vs other groups). AM inhibited viral replication in myocardium at the early and late stage of murine myocarditis effectively and AM combined with taurine enhanced the effect of anti virus. All drugs decreased myocardial damage (P<0.05, vs control group), and the effect of AM combined with taurine was most significant (P<0.05). Conculsion The AM combined with taurine on CVB3 murine myocarditis is more effective than that of AM or taurine.

Microarray analysis of extracellular matrix genes expression in myocardium of mouse with Coxsackie virus B_3 myocarditis

Background Extracellular matrix (ECM) orchestrates cell behaviour including growth, death, apoptosis, adhesion, migration, and invasion by activating several signalling pathways Certain components of ECM, such as integrins, may act as receptors or co-receptors of enterovirus ECM-activated gene expressions in myocardium of viral heart disease including myocarditis and partial cardiomyopathy remain elusive This study was to investigate the expression of ECM-activated genes in myocardium of mouse with viral myocarditis Methods BALB/c mice were infected with Coxsackie virus B 3 (CVB 3) to establish an animal model of myocarditis Uninfected mice were also prepared and served as controls Specific mRNA expression pattern in myocarditic mouse heart was analysed by an in-house cDNA microarray containing 8192 genes Overexpressed ECM genes were selected and subsequently confirmed by Northern blot analysis Results Nine ECM genes were isolated, from the array of 8192 genes, as overexpressed genes in hearts of myocarditic mice in comparison with controls Subsequent Northern blot analysis confirmed that four of the nine genes were highly expressed Expression of these four genes, Fin15, ILk, Lamr1 and ADAMTS-1, has not been reported previously to be induced by Coxsackie virus Conclusion CVB 3-induced myocarditis is associated with gene expression profiles of certain ECM components

Effect of Astragalus membranacaus on Electrophysiological Activities of Acute Experimental Coxsackie B3 Viral Myocarditis in Mice

ＥｆｆｅｃｔｏｆＡｓｔｒａｇａｌｕｓｍｅｍｂｒａｎａｃａｕｓｏｎＥｌｅｃｔｒｏｐｈｙｓｉｏｌｏｇｉｃａｌＡｃｔｉｖｉｔｉｅｓｏｆＡｃｕｔｅＥｘｐｅｒｉｍｅｎｔａｌＣｏｘｓａｃｋｉｅＢ３ＶｉｒａｌＭｙｏｃａｒｄｉｔｉｓｉｎＭｉｃｅＲｕｉＴａｏ（芮涛）；...

牛磺酸对实验性小鼠Coxsackie B3病毒性心肌炎的疗效研究

用Coxsackie B3病毒(CV B3)感染BALB/C小鼠建立实验性病毒心肌炎(VMC)模型,观察了牛磺酸对此模型的作用.结果发现,牛磺酸能减少感染病毒后小鼠的死亡只数,减轻心肌组织病理损害,减少感染CVB3后ECG的异常变化,加快血清及心肌CVB3RNA转阴,感染CVB3后血清牛磺酸浓度明显增加.本实验结果显示,牛磺酸对实验性VMC具有良好的保护作用.

Design,synthesis and biological activity of some novel benzimidazole derivatives against Coxsackie virus B_3

A series of novel benzimidazole derivatives was synthesized and their anti-Coxsackie virus B3 （CVB3） activity was evaluated in VERO ceils. Compounds 9 and 10 exhibited better inhibitory activity than those of ribavirin （RBV） with IC50 values of 5.30 and 1.06 μg/mL, respectively. ？2009 Xian Jin Luo. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

TAURINE INFLUX IN CULTURED RAT CARDIOMYOCYTES AND CHANGES AFTER CVB3 INFECTION

Monolayer culture of spontaneously contracting rat heart cells was obtained from newborn SD rats andseeded intO culture plates. 1OOTCID50 of coxsackie virus B3 (CVB3) (Nancy strain )was added as the infectedgroup. The dynamics of taurine transport across membrane of normal heart cells and changes after infectionwere examined by using radioactive isotope tracing techniques. The results demonstrated that: (1)two uptake systems of taurine with different affinities and capacities were present in the Plasma membrane of cul-tured rat cardiOmyocytes,the K. of high and low affinity were 9. 5 X 10-5mol/l, and l. 3 X 1 O 2 mol/l,,respectively; (2)taurine transport across membrane was sodium-dependent with a positive linear relationshipbetween amount of taurine influx and extracellular concentration of sOdium;in addition,it was alsO regulat-ed by osmOlarity; (3) fyalanine,a carboxylic analog of taurine,could inhibit the uptake of taurine in dosedependently; (4)taurine influx was decreased in cultured rat cardiomyocytes after infected with CVB3. (5)the lnhibition ratio of taurine transport was enhanced as the infected time prolonged. This experiment pro-vides a basis fOr further investigation of kinetic test of taurine transport and more study should be carriedout to learn the effect of taurine in viral myocarditis.

牛磺酸对培养大鼠心肌细胞感染 Coxsackie B_3 病毒的影响

目的：观察牛磺酸对病毒性心肌炎模型的保护作用。方法：取新生ＳＤ大鼠心室肌制备培养搏动心肌细胞，１８ｈ后接种１００ＴＣＩＤ５０的ＣｏｘｓａｃｋｉｅＢ３病毒（ＣＶＢ３）做为实验性病毒性心肌炎模型。结果：牛磺酸能剂量依赖性地减少感染ＣＶＢ３后心肌细胞乳酸脱氢酶、天冬氨酸氨基转移酶的释放，降低上清液的病毒滴度，对搏动停止、细胞病变及超微结构均有保护作用，并发现感染ＣＶＢ３后心肌细胞内牛磺酸含量下降。结论：临床应用牛磺酸治疗病毒性心肌炎有实验依据。

应用嗜鼠心肌Coxsackie B_3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。

柯萨奇B3病毒诱导鼠心肌炎血浆β-内啡肽及其受体的变化

目的：观察病毒性心肌炎病理过程中内源性阿片系统激活的动态变化，探讨阿片肽在心肌炎病理损伤中的作用。方法：以柯萨奇 Ｂ３ 病毒（ Ｃｏｘ Ｂ３）诱导 Ｂ Ａ Ｌ Ｂ／ｃ 鼠心肌炎为实验模型，观察感染后第３，７ ，１０，１５，２５ 和３５ ｄ 鼠体、肝和肺的质量，血浆β内啡肽（β Ｅ Ｐ）水平、心肌细胞膜 β Ｅ Ｐ受体及心肌病损程度。采用放射免疫法测定血浆 β Ｅ Ｐ、放射配体结合法测定β Ｅ Ｐ受体、常规病理检查心肌病损程度。结果：以１ ０００ Ｔ Ｃ Ｉ Ｄ５０ Ｃｏｘ Ｂ３ 感染 Ｂ Ａ Ｌ Ｂ／ｃ 鼠，约９２．８％ （６５～７０）鼠诱导出典型的心肌炎病理改变。感染后 ３～２５ ｄ 受染鼠血浆 β Ｅ Ｐ水平显著高于正常对照组；感染后１０～２５ ｄ 受染鼠肺／体质量和肝／体质量比均高于正常对照鼠，提示受染鼠出现较严重的充血性心力衰竭。同时，受染鼠心肌细胞膜 β Ｅ Ｐ受体 Ｂｍ ａｘ 显著增高， Ｋｄ值变化不明显。结论：由于病毒和自身免疫对心肌细胞的损伤，使病毒性心肌炎患鼠出现充血性心力衰竭，引起循环中阿片肽水平升高及相应受体的变化。通过神经内分泌免疫网络，这些变化必将对心脏的病理损伤产生明显的作用。

川芎嗪对CVB3感染小鼠心肌细胞凋亡的影响

目的探讨川芎嗪对病毒性心肌炎(VM)小鼠心肌细胞凋亡的影响。方法将50只BALB/c小鼠随机分为A、B、C、D、E 5组,每组10只。A组腹腔注射生理盐水0.5 mL;B、C、D、E组均腹腔注射柯萨奇病毒B组(CVB3)0.5 mL诱导小鼠,建立VM模型。1 d后,A、B组给予腹腔注射生理盐水10 mg.kg-1,1次.d-1,连用7 d;C、D、E组分别腹腔注射盐酸川芎嗪注射液5 mg.kg-1、10 mg.kg-1、15 mg.kg-1,1次.d-1,均连用7 d;9 d后处死小鼠。采用TUNEL法检测心肌细胞凋亡;光镜下观察心肌病理变化;免疫组织化学技术检测各组小鼠心肌细胞中Fas/FasL蛋白的表达。结果心肌病理变化:A组无病变;心肌病变积分B组明显高于C、D、E组(均P<0.01);细胞凋亡:与A组比较,B、C、D、E组细胞凋亡指数明显升高(P<0.01或P<0.05),与B组比较C、D、E组细胞凋亡指数明显降低(P<0.01);Fas/FasL蛋白阳性染色指数:B、C、D、E组均显著高于A组(P<0.01或P<0.05),C、D、E组显著低于B组(P<0.01)。结论川芎嗪可通过下调病毒性心肌炎小鼠心肌细胞Fas/FasL蛋白表达,减少心肌细胞凋亡和心肌损伤。

Coxsackie B-3病毒对BALB/C小鼠心肌超微结构的影响

ＢＡＬＢＣ小鼠腹腔注射柯萨奇Ｂ—３病毒后，第三天心肌呈现轻度弥散性病变，大多数细胞结构正常，少数细胞发生病变，心肌闰盘局部扩张，部分线粒体的嵴分解，基质内有少量细小的絮状物质，Ｚ线扭曲．第五天病灶增大，少数心肌细胞结构正常，大部分心肌细胞发生病变．闰盘双膜间隙加宽区域增多，线粒体中絮状高电子密度物质增加，肌原纤维出现肌节病变．至第七天，病灶进一步扩大，整个线粒体遍布钙化的絮状物，肌丝大面积溶解，且细胞间质明显增加．

Clinical and Experimental Study on Huodan Tablet (藿丹片) in Treating Infantile Viral Myocarditis

Objective: To study the effect of Huodan tablet (藿丹片, HDT) in treating infantile viral myocarditis. Methods: Clinical manifestations and physical signs as well as laboratory examinations have been observed. Results: The markedly effective rate was 68%, and total effective rate 91.67% in the treated group, while in the control group, the markedly effective rate was 30.83%, and the total effective rate 70.84%. According to Ridit analysis, significant difference was shown between the two groups. r-treated group=0.5000, r-control group=0.295+2×0.025 respectively. Conclusion: HDT has no toxic side-effect and can be taken safely and conveniently, it conforms to the demands of WHO on new drug for prevention and cure of myocarditis.

清心抗炎饮治疗小儿急性柯萨奇B病毒性心肌炎邪毒侵心证临床研究

目的:观察清心抗炎饮治疗小儿急性柯萨奇B病毒性心肌炎邪毒侵心证的效果。方法:86例按随机数字表法分为两组各43例,对照组给予西药治疗,观察组给予清心抗炎饮治疗。结果:观察组总有效率高于对照组(P<0.05)。治疗后两组证候积分均降低(P<0.05),观察组降低更显著(P<0.05)。治疗后两组血清CK-MB、LDH、CPK水平均下降(P<0.05),且观察组低于对照组(P<0.05)。治疗后两组血清TNF-α、IL-6水平均下降(P<0.05),且观察组低于对照组(P<0.05)。结论:清心抗炎饮治疗急性病毒性心肌炎邪毒侵心证疗效较好。

黄芪甲甙对慢性心肌炎心肌纤维化的影响

目的探讨中药黄芪的活性成分黄芪甲甙对慢性心肌炎心肌纤维化的影响及其相关机制。方法80只Balb/c小鼠随机分为3组,分别为空白对照组(对照组,20只)、慢性心肌炎组(模型组,30只)和黄芪甲甙干预心肌炎组(干预组,30只)。模型组和干预组小鼠每月1次腹腔接种柯萨奇病毒B3(CVB3),对照组腹腔注射等量病毒培养液。对照组和模型组小鼠每日以饮用水喂养,干预组小鼠以加有羧甲基纤维素钠助溶的黄芪甲甙饮用水(浓度300mg/L)喂养。3个月后处死所有存活小鼠,小鼠心脏组织经苦味酸天狼星红染色后以自动图像分析系统计算胶原容积积分(CVF),以Westernblot方法检测各组小鼠心脏组织中转化生长因子β1(TGF-β1)、血小板衍生生长因子BB(PDGF-BB)、基质金属蛋白酶1(MMP-1)及其组织抑制物(TIMP-1)、MMP-13和MMP-14的表达情况。结果黄芪甲甙干预后,与模型组比较,小鼠死亡率降低(53.3%、23.3%,χ2=4.23,P<0.05)、心脏CVF明显下降〔模型组为(17.4±1.2)%、干预组为(8.6±0.9)%,χ2=5.38,P<0.05〕;与对照组比较,TGF-β1、MMP-1和TIMP-1表达减少而MMP-13和MMP-14的表达增多。结论黄芪甲甙可以降低慢性心肌炎小鼠死亡率、减轻其心肌纤维化,其抗纤维化作用是通过抑制TGF-β1、促进MMP-13和MMP-14在心脏中的表达而实现。

抗柯注射液治疗柯萨奇B病毒性心肌炎及其机理研究

目的：探讨抗柯注射液治疗柯萨奇Ｂ病毒性心肌炎作用机理。方法：以苦参中抗柯萨奇病毒的有效成分制成纯度为９９．４３％的抗柯注射液（下称ＲＳＦ）治疗７６例血中柯萨奇Ｂ病毒核酸（ＣＢＶ－ＲＮＡ）持续阳性的病毒性心肌炎患者（Ａ组），并与用葡萄糖加胰岛素加氯化钾治疗的５０例患者（Ｂ组）进行对照。结果：ＲＳＦ对血中ＣＢＶ－ＲＮＡ的清除率与所用药物剂量及血中药物浓度有相关性；患者心悸、胸闷气短的总有效率（９６．０％）、抗。Ｃ律失常的总有效率（１００．０％，４０／４０例）明显优于Ｂ组（６２．５％，２５／４０例）。ＲＳＦ能使射血分数、心脏每搏输出量、心排血量以及心脏指数等心功能指标明显上升（Ｐ＜０．０１），左室重量及重量指数较治疗前明显下降（Ｐ＜０．０５）；ＲＳＦ治疗（５例）５个月后中和抗体恢复正常，ＮＫ细胞活性上升（Ｐ＜０．０１）。结论：ＲＳＦ对逆转病毒性心肌炎的病理状态效果明显。

桑寄生乙醇提取物抗柯萨奇病毒B_3的实验研究

目的 :为研制抗柯萨奇病毒B3 (CVB3 )新药 ,观察了桑寄生乙醇提取物不同部分抗CVB3 药效。方法 :以病毒唑为阳性对照 ,用空斑减数实验检测了抗CVB3 药效。结果 :在HEp 2细胞系统中 ,乙酸乙酯萃取部分 (B)对CVB3 直接杀灭、感染阻断、增殖抑制的ED50 为 2 .32 ,0 .2 4,1.91μg·ml-1。正丁醇萃取部分 (C)的相应数值分别为 1.44 ,2 .0 6 ,3.70 μg·ml-1。病毒唑对CVB3 增殖抑制的ED50 为 7.5 5 μg·ml-1。结论 :桑寄生B部分对CVB3 直接杀灭、感染阻断、增殖抑制的治疗指数 (TI)为 2 2 .6 ,2 19,2 7.5 ,C部分的相应TI为 115 ,16 5 ,2 1.6。病毒唑对CVB3 增殖抑制的TI为 38,数值相近。桑寄生B ,C部分抗CVB3 作用值得开发。

小柴胡汤及其分解剂对柯萨奇B_3m病毒感染乳鼠心肌保护及细胞免疫调节作用的研究

目的 :观察小柴胡汤及其分解剂Ⅰ号和分解剂Ⅱ号对心肌细胞的保护和对机体的免疫调节作用。方法 :以柯萨奇病毒B3亲心肌株 (CVB3m)腹腔注射诱导Balb/c乳鼠心肌炎模型 ,分别予以小柴胡汤、分解剂Ⅰ号及分解剂Ⅱ号 ,经灌胃后 ,于不同时间进行各组的NK细胞活性、T细胞亚群及心肌病理组织学检查。结果 :(1)小柴胡汤和分解剂Ⅱ号在病毒性心肌炎急性期均能明显提高NK细胞活性 ,有调节T细胞亚群功能 ,与病毒对照组比较有显著性差异 (P <0 0 5)。而分解剂Ⅰ号此作用不明显。 (2 ) 3种治疗药物对心肌免疫损伤均有明显保护作用 ,小柴胡汤作用明显优于分解Ⅰ号和分解剂Ⅱ号 (P <0 0 5)。而分解剂Ⅰ号和分解剂Ⅱ号比较未见显著性差异 (P >0 0 5)。结论 :小柴胡汤和分解剂Ⅱ号对病毒性心肌炎NK细胞活性和T细胞亚群具有双向调节作用 ,3种治疗药物对心肌的免疫损伤均有明显保护作用。

高山红景天多糖对病毒感染大鼠心肌细胞的抑制作用

目的：观察高山红景天多糖对柯萨奇Ｂ３病毒感染ＳＤ大鼠心肌细胞的抑制作用。方法：采用原代培养乳鼠心肌细胞的方法，建立实验性病毒性心肌炎模型。结果：高山红景天多糖能明显的抑制体外培养心肌细胞在受到柯萨奇Ｂ３病毒感染后导致的心肌酶释放；显著降低病毒在心肌细胞中的增殖量；其半数有效抑制浓度为１５０ｍｇ·Ｌ－１；有效浓度平均抑制率为７１．３％。结论：高山红景天多糖对在ＳＤ大鼠心肌细胞内增殖的柯萨奇Ｂ３病毒具有一定的抑制作用。

双黄连粉针剂抗柯萨奇病毒B_3型效应的实验研究

目的 探讨双黄连在体外抗柯萨奇病毒 (CVB3)的效应。方法 在Wish细胞上采用微量细胞病变抑制法。结果 (1) 3 0mg/ml、1 5mg/ml双黄连均可直接杀伤 10 0、10TCID50 病毒 ,且 3 0mg/ml直接灭活CVB3 的效应明显。 (2 ) 0 5、0 2 5、0 12 5mg/ml双黄连可分别阻断 10 0、10、1TCID50 病毒吸附细胞。 (3) 1 5、0 2 5、0 2 5mg/ml双黄连可分别抑制 10、5、1TCID50 病毒在胞内复制。结论 在细胞培养上 ,双黄连不仅直接杀伤柯萨奇病毒、阻断或减缓病毒吸附细胞的作用明显 ,而且对细胞内低滴度病毒复制具有一定的抑制作用。