右美托咪定或BNIP3基因siRNA对PC12细胞凋亡的影响
摘要
目的探讨右美托咪啶(DEX)或B淋巴细胞瘤基因(Bcl)-2腺病毒E/B19 kDa相互作用蛋白(BNIP3)基因siRNA对大鼠肾腺嗜铬瘤(PC)12细胞凋亡的影响及机制。方法 PC12细胞分为阴性对照siRNA(NC)组、过氧化氢(H_2O_2)组、DEX组、si-BNIP3组和DEX+si-BNIP3组,其中siRNA转染参照LipofectamineTM2000说明,噻唑蓝(MTT)法检测细胞活力;流式细胞术检测细胞凋亡率及活性氧(ROS)水平;Western印迹检测BNIP3、核因子(NF)-κB p65、κB抑制蛋白激酶(IKK)α和Bcl-2的蛋白表达。结果转染BNIP3 siRNA的PC12细胞BNIP3蛋白表达显著低于空白组(P<0.05)。H_2O_2组细胞活力及Bcl-2的蛋白表达均显著低于NC组,细胞凋亡率、ROS水平及NF-κB p65和IKKα的蛋白表达均显著高于NC组(P<0.05);DEX组和si-BNIP3组细胞活力及Bcl-2的蛋白表达均显著高于H_2O_2组,低于DEX+si-BNIP3组,细胞凋亡率、ROS水平及NF-κB p65和IKKα的蛋白表达均显著低于H_2O_2组,高于DEX+si-BNIP3组(P<0.05)。结论 DEX或BNIP3基因siRNA均可增加PC12活力,抑制细胞凋亡率,联用效果强于单用,机制与降低细胞内ROS水平及下调NF-κB信号有关。
引文
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2 Wang H,Ye Y,Zhu Z,et al.MiR-124 regulates apoptosis and autophagy process in MPTP model of parkinson's disease by targeting to bim[J].Brain Pathol,2016;26(2):167-76.
3 Thanan R,Oikawa S,Hiraku Y,et al.Oxidative stress and its significant roles in neurodegenerative diseases and cancer[J].Int J Mol Sci,2015;16(1):193-217.
4 Lin X,Wu S,Wang Q,et al.Saikosaponin-D reduces H2O2-induced PC12 cell apoptosis by removing ROS and blocking MAPK-dependent oxidative damage[J].Cell Mol Neurobiol,2016;36(8):1-11.
5 Lin L,Lan AP,Zhang LL,et al.The protection of dexmedetomidine against chemical hypoxia-induced neurons injury and its mechanism[J].Guangdong Med J,2012;33(8):1046-9.
6 王琼.右美托咪定对高浓度利多卡因诱导PC12细胞毒性的影响[D].广州:暨南大学,2017.
7 Li Y,Zhang X,Yang J,et al.Methylation of BNIP3 in pancreatic cancer inhibits the induction of mitochondrial-mediated tumor cell apoptosis[J].Oncotarget,2017;8(38):63208-22.
8 Ye F,Wang H,Zhang L,et al.Baicalein induces human osteosarcoma cell line MG-63 apoptosis via ROS-induced BNIP3 expression[J].Tumor Biol,2015;36(6):4731-40.
9 Kahlert W,Maciaczyk D,Dai F,et al.Resistance to hypoxia-induced,BNIP3-mediated cell death contributes to an increase in a CD133-positive cell population in human glioblastomas in vitro[J].J Neuropathol Exp Neurol,2012;71(12):1086-99.
10 Huang H,Hong Q,Tan H,et al.Ferulic acid prevents LPS-induced up-regulation of PDE4B and stimulates the cAMP/CREB signaling pathway in PC12 cells[J].Acta Pharmacologica Sinica,2016;37(12):1543-54.
11 汪军兵,龚正,行妍妍,等.姜黄素对ActD/TNF-α协同诱导PC12细胞损伤的保护作用[J].暨南大学学报(自然科学与医学版),2012;33(4):357-61.
12 张卫东,张浩,汪海,等.右美托咪定对谷氨酸所致PC12细胞损伤的保护作用及其机制[J].南方医科大学学报,2017;37(2):150-6.
13 华潇潇,陈洁琳,张莉莉,等.右美托咪定抑制化学性低氧引起的PC12细胞炎症反应[J].中国药理学通报,2014;30(1):85-9.
14 郑辉哲,王英,张援.右美托咪定对过氧化氢引起的PC12细胞损伤的保护作用[J].中国老年学杂志,2017;37(22):5500-2.
15 He J,Pei L,Jiang H,et al.Chemoresistance of colorectal cancer to 5-fluorouracil is associated with silencing of the BNIP3 gene through aberrant methylation[J].J Cancer,2017;8(7):1187-96.
16 Yuan Y,Zheng Y,Zhang X,et al.BNIP3L/NIX-mediated mitophagy protects against ischemic brain injury independent of PARK2[J].Autophagy,2017;13(10):1754-66.
17 Li R,Yang HQ,Xi HL,et al.Inhibition of CDH17 gene expression via RNA interference reduces proliferation and apoptosis of human MKN28 gastric cancer cells[J].Int J Oncol,2016;50(1):15-22.
18 Wang X,Lu X,Zhu R,et al.Betulinic acid induces apoptosis in differentiated PC12 cells via ROS-mediated mitochondrial pathway[J].Neurochem Res,2017;42(4):1-11.
19 Li S,Du H,Xin D,et al.Raf kinase inhibitor protein regulates oxygen-glucose deprivation-induced PC12 cells apoptosis through the NF-κB and ERK pathways[J].J Clin Biochem Nutr,2016;59(2):86-92.
20 姜恩平,于春荣,李贺,等.北五味子总木脂素对PC12细胞氧化应激损伤的保护作用及其抑制NF-κB/iNOS/NO信号通路的机制[J].吉林大学学报(医学版),2015;41(3):532-6.