目的 考察环状RNA(circRNA)circ_0054633是否靶向miR-375调控过氧化氢(H2O2)诱导的心肌细胞氧化损伤.方法 将H9C2心肌细胞分为对照组(正常培养,未进行任何处理)、H2O2组(150 μmol·L-1 H2O2处理心肌细胞6 h)、H2O2+si-NC 组、H2O2+si-circ_0054633 组、H2O2+miR-NC 组、H2O2+miR-375 组、H2 O2+si-circ_0054633+anti-miR-NC 组、H2 O2+si-circ_0054633+anti-miR-375 组(在心肌细胞中分别转染 si-NC、si-circ_0054633、miR-NC、miR-375 模拟物、si-circ_0054633+anti-miR-NC、si-circ_0054633+anti-miR-375,转染24 h后,用150 μmol·L-1 H2O2处理心肌细胞6 h).用实时荧光定量聚合酶链反应法测定miR-375的表达水平,用试剂盒测定丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,用流式细胞术测定细胞凋亡情况.结果 对照组、H2O2组、H2O2+si-NC 组、H2 O2+si-circ_0054633 组、H2 O2+miR-NC 组、H2O2+miR-375 组、H2O2+si-circ_0054633+anti-miR-NC 组、H2O2+si-circ_0054633+anti-miR-375 组的心肌细胞 miR-375 表达量分别为 1.00±0.00、0.42±0.05、0.40±0.06、0.69±0.08、1.00±0.00、3.41±0.28、1.00±0.00 和 0.26±0.02,MDA 含量分别为(3.19±0.32)、(13.46±1.27)、(15.39±1.04)、(5.26±0.51)、(16.05±1.36)、(9.18±0.85)、(4.89±0.44)和(10.05±0.92)nmol·L-1,SOD 活性分别为(64.69±5.81)、(18.23±1.33)、(17.07±1.41)、(55.74±5.13)、(17.12±1.64)、(47.66±4.59)、(56.77±4.71)和(27.95±2.47)U·mL-1,凋亡率分别为(6.21±0.59)％、(29.22±2.19)％、(30.94±2.85)％、(10.05±0.92)％、(31.14±2.83)％、(13.74±1.24)％、(10.39±0.88)％和(21.31±1.92)％.H2O2 组的上述指标与对照组比较,H2O2+si-circ_0054633组的上述指标与H2O2+si-NC组比较,H2O2+miR-375组的上述指标与H2O2+miR-NC组比较,H2 O2+si-circ_0054633+anti-miR-375 组的上述指标与 H2 O2+si-circ_0054633+anti-miR-NC组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 干扰circ_0054633表达可通过靶向调控miR-375表达,减轻H2O2诱导的心肌细胞氧化应激和凋亡,从而保护心肌细胞免受氧化损伤.
Objective To investigate whether circular RNA(circRNA)circ_0054633 targets microRNA-375(miR-375)to regulate the oxidative damage of cardiomyocytes induced by hydrogen peroxide(H2O2).Methods H9C2 cardiomyocytes were divided into control group(normal culture without any treatment),hydrogen peroxide(H2 O2)group(150 μmol·L-1 H2 O2 treated cardiomyocytes for 6 h),H2O2+si-NC group,H2O2+si-circ_0054633 group,H2O2+miR-NC group,H2O2+miR-375 group,H2O2+si-circ_0054633+anti-miR-NC group,H2O2+si-circ_0054633+anti-miR-375 group(cardiomyocytes were transfected with si-NC,si-circ_0054633,miR-NC,miR-375 mimics,si-circ_0054633+anti-miR-NC,si-circ_0054633+anti-miR-375,respectively;24 h after transfection,cardiomyocytes were treated with 150 μmol·L-1 H2O for 6 h).Real-time fluorescence quantitative polymerase chain reaction was used to determine the expression of miR-375,the content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)were determined by kit,apoptosis was determined by flow cytometry.Results The expression levels of miR-375 in cardiomyocytes of control group,H2O2 group,H2O2+si-NC group,H2O2+si-circ_0054633 group,H2O2+miR-NC group,H2O2+si-circ_0054633+anti-miR-NC group,H2O2+si-circ_0054633+anti-miR-375 group were 1.00±0.00,0.42±0.05,0.40±0.06,0.69±0.08,1.00±0.00,3.41±0.28,1.00±0.00 and 0.26±0.02,respectively;MDA contents were(3.19±0.32),(13.46±1.27),(15.39±1.04),(5.26±0.51),(16.05±1.36),(9.18±0.85),(4.89±0.44)and(10.05±0.92)nmol·L-1,respectively;SOD activities were(64.69±5.81),(18.23±1.33),(17.07±1.41),(55.74±5.13),(17.12±1.64),(47.66±4.59),(56.77±4.71)and(27.95±2.47)U·mL-1,respectively;the apoptosis rates were(6.21±0.59)％,(29.22±2.19)％,(30.94±2.85)％,(10.05±0.92)％,(31.14±2.83)％,(13.74±1.24)％,(10.39±0.88)％and(21.31±1.92)％,respectively.The above indexes of H2O2 group were compared with the control group,the above indexes of H2O2+si-circ_0054633 group was compared with the H2O2+si-NC group,the above indexes of H2O2+miR-375 group was compared with the H2O2+miR-NC group,the above indexes of H2O2+si-circ_0054633+anti-miR-375 group were compared with H2O2+si-circ_0054633+anti-miR-NC group,the differences were statistically significant(all P＜0.05).Conclusion Interfering with the expression of circ_0054633 can reduce the oxidative stress and apoptosis of cardiomyocytes induced by hydrogen peroxide by targeting miR-375,thereby protecting cardiomyocytes from oxidative damage.
王艳炜;吴变稳;曹磊;马杰;封亚丽
河北医科大学第一医院心脏外二科,河北石家庄 050000中国人民解放军联勤保障部队第980医院心血管内科,河北石家庄 050082河北省中医院心血管内二科,河北石家庄 050031||河北省浊毒证重点实验室,河北石家庄 050091
药学
环状RNA_0054633;微小RNA-375;过氧化氢;心肌细胞;氧化应激;凋亡
circular RNA_0054633;microRNA-375;hydrogen peroxide;cardiomyocytes;oxidative stress;apoptosis
《中国临床药理学杂志》 2024 (011)
1560-1564 / 5
河北省中医药管理局科研计划基金资助项目(2018047);河北中医学院教育教学改革研究项目立项课题基金资助项目(22yb-16)
10.13699/j.cnki.1001-6821.2024.11.004
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