摘要
目的:探究下调DNA损伤激活的长链非编码RNA(lncRNA NORAD)抗心房颤动(房颤)大鼠心肌纤维化作用及可能的作用机制。方法:SD大鼠随机分为对照组、房颤组、重组腺相关病毒血清型9阴性对照(rAAV9-NC)组、重组腺相关病毒血清型9载体含用于沉默lncRNA NORAD的小干扰RNA(rAAV9-siNORAD)组、p38丝裂原激活的蛋白激酶(p38MAPK)抑制剂SB203580组、rAAV9-siNORAD+SB203580组,每组8只。建立各组模型大鼠并进行对应干预措施,实验结束后对大鼠进行心电图测试,记录房颤的发生率和持续时间;并检测血清肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)和炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;Masson染色检测心房组织纤维化,计算心房胶原容积分数(CVF);实时荧光定量PCR检测心房组织lncRNA NORAD、Ⅰ型胶原(COL1-A1)和Ⅲ型胶原(COL3-A1)的mRNA表达情况;Western blot检测心房组织丝裂原活化蛋白激酶激酶6(MKK6)、p38MAPK和核因子κB p65(NF-κB p65)及其磷酸化蛋白的表达情况。结果:与对照组相比,房颤组大鼠可观察到P波消失,出现大小不等、不规则的f波,心房组织有明显的纤维化改变,CVF、血清c TnI、CK-MB、TNF-α、IL-6水平、心房组织COL1-A1和COL3-A1的mRNA水平以及lncRNA NORAD表达水平、MKK6蛋白水平、磷酸化p38 MAPK与p38 MAPK的比值和磷酸化NF-κB p65与NF-κB p65比值升高,差异具有统计学意义(P均<0.05);与房颤组相比,rAAV9-siNORAD组和SB203580组大鼠房颤的发生率降低,持续时间缩短(P<0.05),CVF,血清cTnI、CK-MB、TNF-α、IL-6水平,心房组织COL1-A1和COL3-A1的mRNA水平以及lncRNA NORAD表达水平、MKK6蛋白水平、磷酸化p38 MAPK与p38 MAPK的比值及磷酸化NF-κB p65与NF-κB p65的比值降低,差异均具有统计学意义(P均<0.05);分别与单用SB203580和r AAV9-siNORAD相比,两者联合应用后p38 MAPK/NF-κB p65信号通路的激活被进一步抑制,心肌纤维化程度进一步降低。结�
Objectives:To explore the anti-myocardial fibrosis effect and the potential mechanism of down-regulation of long noncoding RNA(lncRNA)noncoding RNA activated by DNA damage(NORAD)in atrial fibrillation(AF)rats.Methods:SD rats were randomly divided into control group,AF group,recombinant adeno-associated virus serotype 9 negative control(rAAV9-NC)group,recombinant adeno-associated virus serotype 9 vector containing small interfering RNA for silencing lncRNA NORAD(rAAV9-siNORAD)group,p38 mitogen activated protein kinase(p38 MAPK)inhibitor SB203580 group,and rAAV9-siNORAD+SB203580 group(n=8 each).Model rats in each group were established and corresponding intervention measures were carried out.After the experiment,the rats were tested by ECG,the incidence and duration of AF were recorded,and the serum creatine kinase isoenzyme MB(CK-MB),troponin I(cTnI)and inflammatory factor tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)level were detected;atrial fibrosis was detected by Masson staining,and atrial collagen volume fraction(CVF)was calculated;real-time fluorescence quantitative PCR was used to detect the expression of lncRNA NORAD,type I collagen(COL1-A1)and typeⅢcollagen(COL3-A1)messenger RNA(mRNA)in atrial tissue;Western blot was used to detect the protein expression of mitogen activated protein kinase kinase 6(MKK6),p38 MAPK and nuclear factorκB p65(NF-κB p65)and phosphorylated NF-κB p65.Results:Compared with the control group,the P wave disappeared in the AF group,and f waves of different sizes and irregular f waves were observed,there were obvious fibrotic changes in the atrial tissue,the CVF level,serum cTnI,CK-MB,TNF-α,IL-6 level,COL1-A1 and COL3-A1 mRNA level in atrial tissue and lncRNA NORAD expression,MKK6 protein expression,p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 ratios were significantly increased,the differences were statistically significant(all P<0.05);compared with the AF group,the incidence of AF in the rAAV9-siNORAD group and the SB203580 group was reduced,the duration was signifi
作者
杨淑玲
谷云飞
杨定宪
高爱玲
尚伟
陈滢伊
张光
YANG Shuling;GU Yunfei;YANG Dingxian;GAO Ailing;SHANG Wei;CHEN Yingyi;ZHANG Guang(Department of Cardiology,Jiaozuo People's Hospital,Jiaozuo(454000),Henan,China)
出处
《中国循环杂志》
CSCD
北大核心
2022年第6期639-646,共8页
Chinese Circulation Journal
基金
河南省医学科技攻关计划联合共建项目(LHGJ20191211)
河南省2022年科技发展计划(222102310076)。
