敲低微小RNA-25表达对缺氧/复氧诱导的H9C2心肌细胞损伤的影响  

Effect of knockdown of microRNA-25 expression on H9C2 cardiomyocyte damage induced by hypoxia/reoxygenation

作　　者：梁车格乐 万全[1] 跟小子 LIANG Che-gele;WAN Quan;GEN Xiao-zi(Department of Cardiology,Affiliated Hospital of Inner Mongolia University,Tongliao 028000,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古民族大学附属医院蒙医心内科,内蒙古通辽028000

出　　处：《中国临床药理学杂志》2022年第6期487-490,共4页The Chinese Journal of Clinical Pharmacology

基　　金：内蒙古自治区自然科学基金资助项目[2017MS(LH)0818]。

摘　　要：目的探讨敲低微小RNA-25(miRNA-25)表达对缺氧/复氧诱导的H9C2心肌细胞损伤的影响。方法将预处理的H9C2心肌细胞分为4组:空白对照组、模型组、阴性组和敲低组。敲低组转染miRNA-25-shRNA干扰质粒0.1μL,空载组转染等量无义干扰序列,空白对照组和模型组分别加入等体积生理盐水。预处理后,除空白对照组外,其他3组细胞构建缺氧/复氧细胞模型。以逆转录-PCR法检测细胞miRNA-25基因表达水平,酶联免疫吸附实验检测培养液乳酸脱氢酶(LDH)水平,CCK-8法检测细胞存活率,流式细胞术检测细胞凋亡情况。结果空白对照组、模型组、阴性组和敲低组的miRNA-25基因表达水平分别为1.31±0.42,0.82±0.33,0.75±0.24和0.27±0.05;这4组的LDH表达水平分别为(49.73±12.54),(109.78±30.74),(112.85±36.44)和(124.63±40.75)U·L^(-1);这4组于72 h的细胞存活率分别为(92.73±6.18)%,(31.82±14.56)%,(29.17±13.38)%和(15.45±6.24)%;这4组的细胞凋亡率分别为(2.37±0.58)%,(46.52±13.56)%,(43.27±8.95)%和(72.78±10.62)%。上述指标:模型组与空白对照组比较,差异均有统计学意义(均P<0.05);阴性组、敲低组与模型组比较,差异均有统计学意义(均P<0.05);敲低组与阴性组比较,差异均有统计学意义(均P<0.05)。结论敲低miRNA-25表达加剧缺氧/复氧诱导的H9C2心肌细胞损伤,可能与加剧细胞炎症反应有关。Objective To investigate the effect of knockdown of miRNA-25 expression on H9C2 cardiomyocyte damage induced by hypoxia/reoxygenation (H/R).Methods H9C2 cardiomyocytes were divided into 4 groups:Blank control group,model group,negative group and knockdown group.Knockdown group with miRNA-25-shRNA 0.1μL interference plasmid and negative group with same amount of nonsense interference sequence,while others with equal volume of saline.After pretreatment,all of them were constructed as H/R model except blank control group.Levels of miRNA-25 gene were detected by reverse transcriptase-PCR.Level of lactate dehydrogenase (LDH) in culture medium were detected by enzyme-linked immunosorbent assay.Cell survival rate were detected by CCK-8 and cell apoptosis rate were detected by flow cytometry.Results The levels of miRNA-25 gene in blank control group,model group,negative group and knockdown group were 1.31±0.42,0.82±0.33,0.75±0.24and 0.27±0.05,respectively;the levels of LDH in four groups were (49.73±12.54),(109.78±30.74),(112.85±36.44) and (124.63±40.75) U·L^(-1);the survival rates of cells at 72 h in four groups were(92.73±6.18)%,(31.82±14.56)%,(29.17±13.38)%and (15.45±6.24)%;the apoptosis rates of four groups were (2.37±0.58)%,(46.52±13.56)%,(43.27±8.95)%and (72.78±10.62)%.Compared between model group and blank control group,the difference of factors were significant (all P<0.05);compared between negative group,knockdown group and model group,the difference of factors were significant (all P<0.05);compared between knockdown group and negative group,the difference of the factors were significant (all P<0.05).ConclusionKnockdown of miRNA-25 expression might influence cellular inflammation to exacerbate H9C2 cardiomyocyte damage with H/R.

关 键 词：微小RNA-25 缺氧/复氧 H9C2心肌细胞 存活率 凋亡率 

分 类 号：R97[医药卫生—药品]