摘要
目的探索右美托咪定(dexmedetomidine,Dex)对心肌细胞缺氧复氧(hypoxia/reoxygenation,H/R)损伤的影响,并探索其可能机制。方法原代培养心肌细胞,右美托咪定处理后,缺氧6 h复氧6 h。细胞分为control组(常氧培养12 h)、H/R组(缺氧6 h复氧6 h)、H/R+Dex组(缺氧前加入1μmol/L右美托咪定)、H/R+Dex+NC组(转染50 nmol/L miR-579-3p inhibitor NC后,进行1μmol/L右美托咪定给药和缺氧复氧处理)、H/R+Dex+inhibitor组(转染50 nmol/L miR-579-3p inhibitor后,进行1μmol/L右美托咪定给药和缺氧复氧处理)。CCK-8实验检测心肌细胞活力,流式细胞仪检测心肌细胞凋亡,Wes-tern blot检测凋亡相关蛋白(Bcl-2,Bax,cleaved Caspase-3)的表达。结果与control组相比,H/R组心肌细胞中miR-579-3p降低,细胞活力降低,凋亡率升高,Bcl-2降低,Bax、cleaved Caspase-3升高(均P<0.05);与H/R组相比,H/R+Dex组miR-579-3p升高,细胞活力升高,凋亡率降低,Bcl-2升高,Bax、cleaved Caspase-3降低(均P<0.05);与H/R+Dex组相比,H/R+Dex+NC组miR-579-3p表达水平、细胞活力、凋亡率、Bcl-2、Bax、cleaved Caspase-3表达水平差异均不明显(P>0.05)。与H/R+Dex组相比,H/R+Dex+inhibitor组miR-579-3p降低,细胞活力降低,凋亡率升高,Bcl-2降低,Bax、cleaved Caspase-3升高(均P<0.05)。结论右美托嘧啶降低了心肌细胞H/R刺激后的凋亡,缓解了心肌细胞缺氧复氧损伤,其保护作用可能是通过上调miR-579-3p来实现的。
Objective To explore the effects of dexmedetomidine(Dex)on cardiomyocyte hypoxia/reoxygenation(H/R)injury,and to explore its possible mechanism.Methods Primary cardiomyocytes were treated with dexmedetomidine,and then hypoxia 6 h reoxygen 6 h.Cardiomyocytes were divided into control group(constant oxygen culture for 12 h),H/R group(hypoxia 6 h reoxygenation 6 h),H/R+Dex group(1 mol/L dexmedetomidine before hypoxia),H/R+Dex+NC group,H/R+Dex+inhibitor group.Cardiomyocytes were transfected with 50 nmol/L miR-579-3p inhibitor NC or 50 nmol/L miR-579-3p inhibitor,and then given 1 mol/L dexmedetomidine before hypoxia in H/R+Dex+NC group and H/R+Dex+inhibitor group,respectively.Cell counting kit-8 was used to detect the cardiomyocyte vitality,flow cytometry was used to detect the apoptosis,Western blot was used to detect the expression levels of apoptosis-related proteins(Bcl-2,Bax,cleaved Caspase-3).Results Compared with control group,miR-579-3p was decreased,cell vitality was decreased,apoptosis rate was increased,Bcl-2 was decreased,and Bax and cleaved Caspase-3 were decreased in H/R group(all P<0.05).Compared with H/R group,miR-579-3p was increased,cell activity was increased,apoptosis rate was decreased,Bcl-2 was increased,and Bax and cleaved Caspase-3 were increased in H/R+Dex group(all P<0.05).There was no significant difference in miR-579-3p expression level,cell activity,apoptosis rate,Bcl-2,Bax,and cleaved Caspase-3 expression levels between H/R+Dex group and H/R+Dex+NC group(all P>0.05).Compared with H/R+Dex group,miR-579-3p was decreased,cell activity was decreased,apoptosis rate was increased,Bcl-2 was decreased,and Bax and cleaved Caspase-3 were decreased in H/R+Dex+inhibitor group(all P<0.05).Conclusion Dexmedetomidine can decrease the cardiomyocyte apoptosis after H/R treatment,and alleviate H/R injury of cardiomyocyte.The protective effect may be achieved by increasing the expression of miR-579-3p.
作者
姜玉玉
尹天玥
李小雨
于汝
程向阳
JIANG Yuyu;YIN Tianyue;LI Xiaoyu;YU Ru;CHENG Xiangyang(Department of Anesthesiology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233030,China)
出处
《山西医科大学学报》
CAS
2021年第1期44-49,共6页
Journal of Shanxi Medical University
基金
安徽省教育厅自然科学研究重点项目(KJ2018A0212)
国家级大学生创业立项项目(202010367012)。
