摘要
目的探讨长链非编码RNA ZFAS1(long non-coding RNA ZFAS1,lncZFAS1)对心肌细胞纤维化发生发展的影响。方法用转化生长因子-β(transforming growth factor-β,TGF-β)诱导HL-1小鼠心肌细胞建立心肌纤维化模型。将lncZFAS1过表达载体或小干扰RNA(siRNA)转染至该模型中。通过蛋白质印迹法检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ⅰ型胶原蛋白(collagenⅠ)水平,将lncZFAS1过表达、干扰片段转染到心肌细胞纤维化模型中,通过实时荧光定量PCR检测lncZFAS1的转录水平。通过CCK-8法检测细胞增殖活力,采用流式细胞术检测细胞凋亡率,通过细胞免疫荧光检测LC3自噬斑形成情况。结果与对照组相比,TGF-β诱导组α-SMA、collagenⅠ、LC3 B蛋白表达显著升高,p62蛋白表达显著降低;lncZFAS1表达量显著增加;细胞活力和自噬斑数量显著增加;细胞凋亡率显著降低(均P<0.05)。模型干扰lncZFAS1后,lncZFAS1表达量降低,细胞活力、自噬斑数量减低,凋亡率升高,p62蛋白表达增加,LC3 B蛋白表达降低,且差异均有统计学意义(均P<0.05)。过表达lncZFAS1后则表现出相反的结果。结论lncZFAS1通过调控细胞自噬,促进心肌细胞纤维化的发生与发展。靶向抑制lncZFAS1可能为延缓心血管疾病进展提供新的治疗策略。
Objective To investigate the impact of long non-coding RNA ZFAS1(lncZFAS1)on the initiation and progression of cardiomyocyte fibrosis.Methods HL-1 mouse cardiac myocytes were induced by transforming growth factor-β(TGF-β)to establish a cardiomyocyte fibrosis model.LncZFAS1 overexpression vectors or small interfering RNA(siRNA)were transfected into the model.Protein levels ofα-smooth muscle actin(α-SMA)and collagenⅠwere detected by western blotting;lncZFAS1 overexpression and interference fragments were transfected into the cardiomyocyte fibrotic model.The transcriptional level of lncZFAS1 was measured via real-time quantitative PCR.Cell proliferation viability was assessed using the CCK-8 assay,apoptosis rate was analyzed by flow cytometry,and LC3 autophagy puncta formation was examined using cellular immunofluorescence.Results Compared with the control group,the TGF-βinduction group showed significantly increased protein expression ofα-SMA,collagenⅠand LC3 B,significantly decreased p62 protein expression,elevated lncZFAS1 expression,enhanced cell viability and autophagy puncta formation,and reduced apoptosis rate(all P<0.05).After interfering with lncZFAS1 in the model,it showed a decrease in the expression level of lncZFAS1,a decrease in cell viability and the number of autophagy puncta,an increase of apoptosis rate and expression of p62 protein,and a decrease in the expression of LC3 B protein,all with statistically significant differences(all P<0.05).However,after overexpression of lncZFAS1,the opposite results were presented.Conclusion LncZFAS1 promotes the initiation and progression of cardiomyocyte fibrosis by regulating cellular autophagy.Targeted inhibition of lncZFAS1 may provide a novel therapeutic strategy for delaying the progression of cardiovascular diseases.
作者
林小花
欧阳煜
程效
肖婧雯
江宇
魏沁菲
张彦
LIN Xiaohua;OUYANG Yu;CHENG Xiao;XIAO Jingwen;JIANG Yu;WEI Qinfei;ZHANG Yan(Department of Cardiology,Fuzhou First General Hospital(Fuzhou Institute of Cardiovascular Diseases),Fuzhou Fujian 350009,China)
出处
《实用心电与临床诊疗》
2025年第4期494-500,共7页
PRACTICAL ELECTROCARDIOLOGY AND CLINICAL TREATMENT
基金
福州市科技计划项目(2022-S-024)
福州市“十四五”临床专科培强培优培育建设项目(20220101)。
作者简介
林小花,主要从事心血管疾病相关研究;通信作者:张彦,E-mail:18558750600@163.com。
