丙泊酚抑制缺氧/复氧HUVECs细胞中miR-15b表达及细胞凋亡的实验研究
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摘要
目的:研究丙泊酚在缺氧/复氧细胞模型中对微小RNA-15b(miR-15b)及血管内皮细胞凋亡的影响。方法:建立缺氧/复氧细胞模型[人脐静脉内皮细胞(HUVECs)],设置实验组、对照组、空白组,其中实验组HUVECs缺氧/复氧培养24h后,加入丙泊酚(150μmol/L)干预培养6h;实时定量PCR(RT-qPCR)检测各组HUVECs中miR-15b的表达情况;RT-qPCR、Western-blot检测各组HUVECs中miR-15b靶基因B淋巴细胞瘤-2(bcl-2)的表达情况;流式细胞术检测各组细胞凋亡能力。结果:RT-qPCR结果显示,实验组miR-15b的表达水平低于对照组(P<0.05);RT-qPCR及Western-blot结果显示,实验组bcl-2在mRNA和蛋白水平的表达均高于对照组(P<0.05);实验组HUVECs的细胞凋亡率低于对照组(P<0.05)。结论:丙泊酚能够抑制缺氧/复氧HUVECs细胞中miR-15b的表达,并影响其靶基因抗凋亡蛋白bcl-2的表达,从而降低细胞凋亡率。
Objective:To investigate the effect of Propofol on expression of microRNA-15 b(miR-15 b)and vascular endothelial cells apoptosis in hypoxia/reoxygenation cell model.Methods:The hypoxia/reoxygenation cell model[human umbilical vein endothelial cells(HUVECs)]was established.The experimental group,the control group and the blank group were set up.HUVECs were incubated with hypoxia/reoxygenation for 24 h,then treated with 150μmol/L Propofol for 6 h.The expression of miR-15 bin each group of HUVECs was detected by real-time quantitative PCR(RT-qPCR).The expression of B lymphoblastoma-2(bcl-2)expression were detected by RTqPCR and Western-blot.Flow cytometry was used to detect apoptosis in each group.Results:RT-qPCR results showed that the expression level of miR-15 bin the experimental group was lower than that in the control group(P<0.05).The results of RT-qPCR and Western-blot showed that the expression of bcl-2 mRNA and protein in the experimental group were higher than that in the control(P<0.05).The apoptosis rate of HUVECs in experimental group was lower than that in control group(P<0.05).Conclusion:Propofol can inhibit the expression of miR-15 b in hypoxia/reoxygenation HUVECs and affect the expression of its target gene bcl-2(anti-apoptotic protein,thus reducing the apoptosis rate.
Objective:To investigate the effect of Propofol on expression of microRNA-15 b(miR-15 b)and vascular endothelial cells apoptosis in hypoxia/reoxygenation cell model.Methods:The hypoxia/reoxygenation cell model[human umbilical vein endothelial cells(HUVECs)]was established.The experimental group,the control group and the blank group were set up.HUVECs were incubated with hypoxia/reoxygenation for 24 h,then treated with 150μmol/L Propofol for 6 h.The expression of miR-15 bin each group of HUVECs was detected by real-time quantitative PCR(RT-qPCR).The expression of B lymphoblastoma-2(bcl-2)expression were detected by RTqPCR and Western-blot.Flow cytometry was used to detect apoptosis in each group.Results:RT-qPCR results showed that the expression level of miR-15 bin the experimental group was lower than that in the control group(P<0.05).The results of RT-qPCR and Western-blot showed that the expression of bcl-2 mRNA and protein in the experimental group were higher than that in the control(P<0.05).The apoptosis rate of HUVECs in experimental group was lower than that in control group(P<0.05).Conclusion:Propofol can inhibit the expression of miR-15 b in hypoxia/reoxygenation HUVECs and affect the expression of its target gene bcl-2(anti-apoptotic protein,thus reducing the apoptosis rate.
引文
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[15]金丹,周烈,刘启祥,等.miR-15b在恶性黑色素瘤A375细胞中的表达及其对细胞转移和侵袭的影响[J].医学研究杂志,2017,46(7):156-159.
[16]高伟,张硕,梁左迪,等.右美托咪定预处理对内毒素血症大鼠海马区Bcl-2、Bax表达及认知功能的影响[J].国际麻醉学与复苏杂志,2017,38(9):773-776.
[2]常建华,张世平,王臻,等.丙泊酚联合七氟烷对扁桃体切除术患儿麻醉苏醒期血流动力学及躁动情绪影响研究[J].陕西医学杂志,2018,47(7):908-911.
[3]Ha JH,Noh HS,Shin IW,et al.Mitigation of H2O2-induced autophagic cell death by propofol in H9c2cardiomyocytes[J].Cell Biol Toxicol,2012,28(1):19-29.
[4]赖永旭,李艳萍.MicroRNA-184靶向作用EPB41L5抑制肾细胞癌存活与迁移研究[J].陕西医学杂志,2017,46(3):294-297.
[5]周斌,徐国海,罗振中.miRNA与神经病理性疼痛的关系研究进展[J].临床麻醉学杂志,2014,30(9):931-933.
[6]刘军政,张书艳,高建强.骨髓间充质干细胞对软骨诱导分化过程中MicroRNA调控机制[J].陕西医学杂志,2017,46(8):989-991.
[7]Yin C,Wang X,Kukreja CR,et al.Endogenous microR-NAs induced by heat-shock reduce myocardial infarction following ischemia-reperfusion in mice[J].FEBS Letters,2008,582(30):4137-4142.
[8]Chen Z,Hu Z,Liu Z,et al.Differential microRNA profiling in a cellular hypoxia reoxygenation model upon posthypoxic propofol treatment reveals alterations in autophagy signaling network[J].Oxid Med Cell Longev,2013,2013(6):378-484.
[9]黄玉民.MicroRNA-15b下调Bcl-2促进耐药肺癌细胞凋亡[J].临床肺科杂志,2013,18(11):2070-2072.
[10]卢悦,王晶晶,梁广彬,等.microRNA20b调节PPARγ抑制缺氧/复氧诱导的内皮细胞凋亡[J].中国中西医结合外科杂志,2017,23(3):283-286.
[11]胡喆,梁就庆,王晶晶,等.丙泊酚通过诱导自噬保护血管紧张素Ⅱ引起细胞损伤[J].广东医科大学学报,2017,35(4):333-341.
[12]徐桂萍,后晓超.丙泊酚与七氟醚对结直肠癌根治术患者缺氧诱导因子-1水平的影响[J].国际麻醉学与复苏杂志,2017,38(11):967-992.
[13]Noh HS,Shin IW,Ha JH,et al.Propofol protects the autophagic cell death induced by the ischemia_reperfusion injury in rats[J].Mol Cells,2010,30(5):455-460.
[14]Delbridge LMD,Mellor KM,Taylor DJ,et al.Myocardial stress and autophagy:mechanisms and potential therapies[J].Nat Rev Cardiol,2017,14(7):412-425.
[15]金丹,周烈,刘启祥,等.miR-15b在恶性黑色素瘤A375细胞中的表达及其对细胞转移和侵袭的影响[J].医学研究杂志,2017,46(7):156-159.
[16]高伟,张硕,梁左迪,等.右美托咪定预处理对内毒素血症大鼠海马区Bcl-2、Bax表达及认知功能的影响[J].国际麻醉学与复苏杂志,2017,38(9):773-776.