ERK1/ET1在慢性心肌炎病毒持续感染与纤维化作用及清心Ⅱ号干预机制  

ERK1 and ET1 in Viral Myocarditis in Myocardial Fibrosis Formation Effect and Qingxin Ⅱ Intervention

作　　者：刘旺[1] 王娟 徐百鸿[3] 程志清[4] 

机构地区：[1]杭州市肿瘤医院,浙江杭州310003 [2]杭州市儿童医院,浙江杭州310014 [3]濮阳市中医院,河南濮阳457001 [4]浙江中医药大学,浙江杭州310053

出　　处：《中华中医药学刊》2017年第5期1250-1253,I0014,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药科技计划项目(22010ZB122)

摘　　要：目的:ERK1/ET1在慢性心肌炎病毒持续感染与纤维化表达及清心Ⅱ号干预作用。方法:150只清洁级体重14~16 g健康雄性Balb/c小鼠,采用随机数字表法分出10只作为正常组,其余140只采用腹腔注射含不同浓度柯莎奇病毒B3半数组织培养感染剂量=10-5(coxsackie virus B3tissue culture infective dose50=10-5,CVB3 TCID50=10-5)的病毒液3次,每次每只2 m L,首次1:2000浓度CVB3病毒液,2周后腹腔接种1∶1600浓度CVB3病毒液,4周后腹腔注射1∶800浓度CVB3病毒液,60天后模型构建成功,存活小鼠94只。随机选取90只,分为清心II号高、中、低剂量组各20只,卡托普利组20只,模型组10只。正常组和模型组给予生理盐水灌胃,治疗组分别给予卡托普利和清心II号高中低剂量灌胃。实验结束,摘眼球取血后颈椎脱臼法处死动物,采集心脏组织液氮速冻。用放射免疫法测各组血浆内皮素1(endothelium 1,ET1)含量;RT-PCR病毒、P-ERK1基因检测;MASSON染色观察各组纤维化形成情况,并检测胶原阳性面积占测量的面积的百分比的即容积分数。结果:造模后ET1、ERK1、CVB3RNA模型组及各治疗组均高于正常组(P<0.05);MASSON染色后正常组含少量胶原纤维,分布于血管周围;模型组心肌细胞排列紊乱,胶原纤维与正常组比较增生明显(P<0.05);治疗后各治疗组ERK1、ET1、CVB3RNA与模型组比较相应均减低(P<0.05),P-ERK1各治疗组均接近正常组(P>0.05),各治疗组胶原纤维较模型组减少(P<0.05),清心Ⅱ号各治疗组随清心Ⅱ号剂量增加ERK1、ET1、CVB3RNA逐渐减少,高剂量组胶原含量接近于正常组(P>0.05),清心Ⅱ号高中低剂量组与卡托普利组疗效相当(P>0.05)。结论:在慢性病毒性心肌炎心肌细胞中CVB3RNA持续存在,ET1与ERK1在心肌病毒持续感染及心肌纤维化形成过程中发挥重要作用,清心Ⅱ号可显著减少ET1、ERK1在心肌的表达,且具有清除柯莎奇病毒及显著抗心肌纤维化作用。Objective ： To observe the effect of ERK1 and ET1 effect on viral myocarditis myocardial fibrosis formation and the intervention effect of Qingxin II Formula. Methods ： A total of 150 health male Balb/c mice were randomly divided into two groups： control group （ n = 10） and experiment group （ n = 140）. The mice of experiment group were injected coxsackie virus B3 （CVB3） （TCID50 = 10 -5 ） poison intraperitoneally. The 2mL CVB3 poison was injected intraperitoneally for each mouse, repeating tree times. The concentration and time were injected as following： 1 ： 2000 of CVB3 at beginning, 1 ： 6000 of CVB3 after two weeks and 1 ： 8000 of CVB3 4 weeks later. The model was constructed successfully and 94 mice were alive. Randomly select 90 for following three groups： model group （ n = 10） and captopril group （ n = 20） and Qingxin II high, medium and low dose groups （ n = 20）. The normal group and model group were fed normal saline by garage. Experiment groups were fed captopril and Qingxin II groups （high, medium and low dose）, respective- ly. The mice were killed by cervical dislocation after heart collected and snap -frozen in liquid nitrogen. At the end of the experiment, use cervical dislocation method to execute animals after taking out eyeball blood. Collect liquid nitrogen frozen heart tissue. By RIA method measure groups＇ plasma endothelinm 1 （ET1） content. Usw RT - PCR virus and P - ERK1 genetic testing. MASSON staining was used to observe each group＇ s fibrosis formation and examine collagen of positive area percentage of the volume fraction. Results After model constructed, ETI, ERK1 and CVB3 RNA of model group and treatment group were higher than those in normal group （ P 〈 0. 05 ）. After MASSON staining, normal group contained a small amount of collagen fibers, distributed in the blood vessels. Model group disordered arrangement of cardiac muscle cells and collagen fiber hyperplasia compared with the normal group （ P 〈 0.05 ）. After treatm

关 键 词：ET1 P-ERK1 病毒基因检测 MASSON染色 心肌纤维化 

分 类 号：R285.5[医药卫生—中药学]