CD40小干扰RNA对自身免疫性心肌炎大鼠血白细胞介素-21与白细胞介素-35表达的影响  

Effect of CD40 small interfering RNA on the expressions of peripheral blood interleukin -21 and interleukin - 35 in rats with autoimmune myocarditis

作　　者：杨德娟[1] 韩波[1] 梁燕[1] 朱梅[2] 高聆[3] 丁国玉[4] 张荣军[4] 石鑫淼 房慧娟[1] 

机构地区：[1]山东大学附属省立医院小儿心脏科,济南250000 [2]山东大学附属省立医院心脏超声科,济南250000 [3]山东大学附属省立医院中心实验室,济南250000 [4]山东省千佛山医院儿科,济南250014

出　　处：《中华实用儿科临床杂志》2016年第13期1021-1026,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：山东省科技发展计划（2013GSF11861）;济南市科技发展计划（201401262）

摘　　要：目的探讨CD40小干扰RNA（siRNA）对实验性自身免疫性心肌炎（EAM）大鼠外周血白细胞介素-21（IL-21）与白细胞介素-35（IL-35）表达的影响及意义。方法将6～8周20只雄性Lewis大鼠采用随机数字表法分为正常组、EAM组、CD40 siRNA组及siRNA组，每组5只。实验第0天及第7天，正常组大鼠双后肢足垫区皮下注射PBS缓冲液，其余3组双后肢足垫区皮下注射猪心肌球蛋白，建立EAM大鼠模型。第7天CD。siRNA组及siRNA组大鼠经尾静脉分别注射CD40 siRNA及siRNA慢病毒表达载体。第21天，行超声心动图检查后处死大鼠。光镜下观察心肌组织病理变化并计算心肌组织病理积分。采用酶联免疫吸附试验检测大鼠外周血IL-21与IL-35水平。结果1．除正常组大鼠，余3组大鼠EAM发病率均为100％，无一例死亡。2．CD40 siRNA组大鼠心脏质量／体质量比值明显低于EAM组，差异有统计学意义（3．13±0．21比3．80±0．29，F=0．332，P〈0．05）；其心肌组织病理积分亦明显低于EAM组，差异有统计学意义（2．22±0．43比3．32±0．51，F=0．456，P〈0．05）。3．超声心动图检测显示，EAM组有1例大鼠探及大量心包积液，CD40 siRNA组大鼠均未探及心包积液。EAM组、CD40 siRNA组及siRNA组大鼠室间隔和左心室后壁均增厚，心腔缩小，室壁运动减低。CD。siRNA组左心室短轴缩短率较EAM组明显升高，差异有统计学意义[（63．34±11．06）％比（38．56±6．98）％，F=16．080，P〈0．05]。4．CD。siRNA组大鼠外周血IL-21水平明显低于EAM组，差异有统计学意义[（141．19±17．46）ng／L比（157．81±17．58）ng／L，F=57．008，P〈0．05]；而其IL-35水平明显高于EAM组，差异有统计学意义[（195．96±18．26）ng／L比（174．78±13．91）ng／L，F=31．727，P〈0．05]。5．EAM组大鼠外周血IL-21水平与心肌组织病理积分呈正相关（r=0．69，P〈0．05），而其IL-35水平Objective To explore the effect of CD40 small interfering RNA（siRNA） on the expressions of pe- ripheral blood interleukin（IL） -21 and IL- 35 in rats with experimental autoimmune myocarditis （EAM） and its sig- nificance. Methods Twenty 6 - 8 week male Lewis rats were divided into normal group, EAM group, CD40 siRNA group and siRNA group by using random number table, with 5 rats in each group. The normal rats were induced with phos- phate buffer saline in double foot pads on day 0 and day 7, while the rest 3 groups were induced with cardiac myosin protein to establish EAM models. The rats in CD40siRNA group and siRNA group were respectively injected with CD40 siRNA and siRNA slow virus expression vector through the tail vein of rats on day 7. The rats were executed on 21 day after echocardiogram examination was made. The histopathologic changes were observed by using light microscope and the myocardial histopathology scores were calculated. Enzyme - linked immunosorbent assay was used to determine the levels of IL - 21 and IL - 35 in peripheral blood. Results （ 1 ） Except the normal group, the total incidence rate of rats of each group was 100% ,and there was no rat death. （2）Compared with EAM group,the heart mass/body ratio and myocardial histopathology scores were lower in CD40 siRNA group, and the differences were significant （3.13 ± 0.21 vs 3.80±0.29,2.22 ±0.43 vs 3.32 ±0.51 ,F =0. 332,0. 456,all P 〈 0.05）. （3） The echocardiogram showed that there was only 1 rat in EAM group with massive pericardial effusion, and there was no pericardial effusion in CD40 siRNA group. EAM group, CD40 siRNA group and siRNA group displayed hypertrophy of the ventricular septum and left ventricular wall, narrow heart cavity and weakening of ventricular wall motion. The left ventrieular shortening rate in CD4o siRNA group was significantly higher than that in the EAM group [ （63.34 ± 11.06） % vs （ 38.56 ± 6.98 ） %, F = 16. 080 ,P 〈 0.05 ]. （4）The peripheral blood level of IL -21 in

关 键 词：CD40小干扰RNA 自身免疫性心肌炎 大鼠 白细胞介素-21 白细胞介素-35 

分 类 号：R542.21[医药卫生—心血管疾病]