摘要
目的:探讨沉默长链非编码RNA染色体X失活特异转录物(lncRNA XIST)通过转换生长因子β1(TGF-β1)/Smad家族成员3(Smad2)通路对扩张型心肌病(DCM)大鼠心肌纤维化的影响。方法:选择SD大鼠64只,分为对照组、模型组、sh-NC组、sh-XIST组,每组各16只,除对照组外,其余组建立DCM大鼠模型,sh-NC组、sh-XIST组分别经尾静脉注射空载体、lncRNA XIST小干扰RNA(siRNA)载体溶液,对照组、模型组以等量0.9%氯化钠溶液尾静脉注射,每周1次,连续4周,末次干预后48 h,以超声仪器测各组大鼠心功能,以苏木精-伊红(HE)染色观察心脏组织结构变化,TUNEL法检测心肌细胞凋亡,荧光定量-聚合酶链反应(RT-qPCR)、蛋白质免疫印迹(Western blot)对心肌组织中TGF-β1、Smad2蛋白表达进行测定。结果:与对照组比较,模型组、sh-NC组心功能恶化,sh-XIST组心功能改善(均P<0.05)。模型组、sh-NC组大鼠心肌细胞排列不规则,间质增宽,并有坏死、肥大出现,且有纤维组织增生、胶原沉积情况,sh-XIST组大鼠心肌细胞排列较规则,未见明显坏死情况,胶原沉积、纤维化有改善。心肌组织中lncRNA XIST mRNA水平从高到低依次为模型组、sh-NC组、sh-XIST组、对照组(均P<0.05),与对照组比较,模型组、sh-NC组心肌组织中TGF-β1、Smad2、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(CoI-Ⅰ)蛋白水平升高,sh-XIST组低于模型组,但高于对照组(均P<0.05)。结论:沉默lncRNA XIST可减轻DCM大鼠心肌细胞凋亡,改善心肌纤维化状态,其作用机可能与TGF-β1/Smad2通路有关。
Objective:To investigate the effect of silencing long non-coding RNA chromosome X inactivating specific transcript(lncRNA XIST)on myocardial fibrosis in rats with dilated cardiomyopathy(DCM)through transforming growth factor-β1(TGF-β1)/Smad family member 3(Smad2)pathway.Methods:Sixty-four SD rats were divided into control group,model group,sh-NC group and sh-XIST group,with 16 rats in each group.DCM rat model was established in the other groups except the control group.The empty vector and lncRNA XIST small interfering RNA(siRNA)vector solution were injected into the tail vein in the sh-NC group and sh-XIST group,respectively.The control group and the model group were injected with the same amount of 0.9%sodium chloride solution via tail vein once a week for 4 weeks.Forty-eight hours after the last intervention,the cardiac function was measured by ultrasonic instrument,the structural changes of cardiac tissue were observed by hematoxylin-eosin(HE)staining,and the apoptosis of myocardial cells was detected by TUNEL method.Real-time quantitative polymerase chain reaction(RT-qPCR)and Western Blot were used to detect the mRNA and protein expression of TGF-β1 and Smad2 in myocardial tissue.Results:Compared with the control group,the cardiac function was deteriorated in the model and sh-NC groups,and was improved in the sh-XIST group(all P<0.05).In the model group and the sh-NC group,the myocardial cells were arranged in disorder,cell necrosis and hypertrophy were observed,the intercellular matrix was widened,a large number of collagen deposition and fibrous tissue proliferation were observed,and the myocardial cells were arranged in order,there was no obvious necrotic cells,and collagen deposition and fibrosis were significantly reduced in the sh-XIST group.The mRNA level of lncRNA XIST in myocardial tissue from high to low was the model group,the sh-NC group,the sh-XIST group,and the control group(all P<0.05).Compared with the control group,the levels of TGF-β1,Smad2,α-smooth muscle actin(α-SMA)and collagen typeⅠ(CoI-Ⅰ)protein in myocardial tissue were increased in the model group and the sh-NC group,and those in the sh-XIST group were lower than those in the model group,but higher than those in the control group(all P<0.05).Conclusion:Silencing lncRNA XIST can reduce myocardial cell apoptosis and improve myocardial fibrosis in DCM rats,which may be related to the TGF-β1/Smad2 pathway。
作者
刘佳
刘丽蒙
赵丽
郝恒瑞
纪美霞
LIU Jia;LIU Limeng;ZHAO Li;HAO Hengrui;JI Meixia(Department of Cardiac Surgery,Affiliated Hospital of Xingtai Medical College,Xingtai 054000,China;Department of Cardiac Surgery,Xingtai Central Hospital,Xingtai 054000,China;Department of Pediatrics,Xingtai People’s Hospital,Hebei Medical University,Xingtai 054099,China;Department of Ultrasound,Xingtai People’s Hospital,Hebei Medical University,Xingtai 054099,China)
出处
《陕西医学杂志》
2025年第7期914-918,共5页
Shaanxi Medical Journal
基金
河北省中医药管理局科研项目(2021453)。
作者简介
第一作者:刘佳,学士,主管护师,从事心外科相关疾病研究,E-mail:yqxc527@163.com;通信作者:纪美霞,学士,主任医师,从事超声医学相关研究,E-mail:haohengrui2024@sina.com。
