|本期目录/Table of Contents|

[1]汤 娅,石 蓓,赵然尊.过表达Nrf2对氧化应激状态下骨髓间充质干细胞凋亡的保护作用[J].遵义医科大学学报,2019,42(05):509-516.
 Tang Ya,Shi Bei,Zhao Ranzun.Protective effect of Nrf2 overexpression on apoptosis of bone marrow mesenchymal stem cells under oxidative stress[J].Journal of Zunyi Medical University,2019,42(05):509-516.
点击复制

过表达Nrf2对氧化应激状态下骨髓间充质干细胞凋亡的保护作用()
     
分享到:

《遵义医科大学学报》[ISSN:1000-9035/CN:22-1262/O4]

卷:
第42卷
期数:
2019年05期
页码:
509-516
栏目:
基础医学研究
出版日期:
2019-10-25

文章信息/Info

Title:
Protective effect of Nrf2 overexpression on apoptosis of bone marrow mesenchymal stem cells under oxidative stress
文章编号:
1000-2715(2019)05-0509-08
作者:
汤 娅石 蓓赵然尊
(遵义医科大学附属医院 心血管内科,贵州 遵义 563099)
Author(s):
Tang YaShi BeiZhao Ranzun
(Department of Cardiology,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China)
关键词:
Nrf2 骨髓间充质干细胞 氧化应激 细胞凋亡
Keywords:
Nrf2 bone marrow mesenchymal stem cells oxidative stress cell apoptosis
分类号:
R282.71
DOI:
-
文献标志码:
A
摘要:
目的 探讨Nrf2调节氧化应激状态下对骨髓间充质干细胞(Bone marrowmesenchymal stem cells,BMSCs )凋亡的作用及机制。方法 原代培养SD大鼠BMSCs; 构建过表达Nrf2重组慢病毒载体(pLV-hefla-EGFP)和沉默Nrf2重组慢病毒载体(pLV-Hu6-Nfe212 shRNA02-hEFLa-EGFP),并以空载体质粒pLV-hefla-EGFP、基因沉默质粒pLV-Hu6 -shRNA02-hEFla-EGFP作为实验空载体阴性对照,分别转染骨髓间充质干细胞,建立稳定过表达Nrf2及沉默Nrf2的骨髓间充质干细胞系; H2O2模拟心肌梗死后氧化应激状态; 随机分为5组:空白对照组(Control)、Nrf2干扰空载体组(Nrf2-Vector)、Nrf2干扰组(si-Nrf2)、Nrf2过表达空载体组(Nrf2-Scramble)、Nrf2过表达组(LV-Nrf2)。Western Blot鉴定慢病毒转染Nrf2 BMSCs成功; Annexin V- FITC/PI双标记流式细胞术检测BMSCs凋亡率; Western Blot检测Nrf2、HO-1、NQO1等抗氧化基因蛋白的表达。结果 Control组、si-Nrf2组、LV-Nrf2组总体凋亡率(%)分别为68.52±0.04、89.22±0.02、38.99±0.06; 与Control组相比,LV-Nrf2组BMSCs的总体凋亡率显著降低(P<0.05),si-Nrf2组BMSCs总体凋亡率显著升高(P<0.05); 此外,与Control组相比, LV-Nrf2组抗氧化相关蛋白Nrf2、HO-1、NQO1相对表达量显著上调(P<0.05); si-Nrf2组抗氧化相关蛋白Nrf2、HO-1、NQO1相对表达量显著下调(P<0.05)。结论 过表达Nrf2可有效减少氧化应激状态下骨髓间充质干细胞的凋亡,其机制可能是通过激活Nrf2/HO-1/NQO1信号通路。
Abstract:
ObjectiveTo investigate the mechanisms underlying Nrf2 regulated apoptosis of Bone Marrow Mesenchymal Stem Cells(BMSCs)under oxidative stress.Methods Primary BMSCs culture was from SD rat.Lentivirus transfected BMSCs was established.The oxidative stress state after myocardial infarction was stimulated by H2O2.This study was randomly divided into five groups: Control group; the overexpression of Nrf2 group(LV-Nrf2); the empty vector overexpressed of Nrf2 group(Nrf2-Scramble); the interference of Nrf2 group(si-Nrf2); the interfered with empty Vector of Nrf2 group(Nrf2-Vector).Western blot assay identified the transfection of Nrf2 BMSCs with lentivirus.Annexin V -FITC/PI double-labeled flow cytometry was used to detect the apoptosis rate of BMSCs.Western blot assay was used to detect the expression of Nrf2,HO-1,NQO1 and other related anti-oxidant proteins.Results Compared with Control group,the overall apoptosis rate of BMSCs in LV-Nrf2 group was significantly reduced.The overall apoptosis rate of BMSCs in si-Nrf2 group was increased.In addition,compared with Control group,the expression levels of anti-oxidant genes and related proteins Nrf2,HO-1 and NQO1 in LV-Nrf2 group were increased.The expression levels of anti-oxidant genes and related proteins Nrf2,HO-1 and NQO1 in si-Nrf2 group were decreased.Conclusion Overexpression of Nrf2 could effectively reduce the apoptosis of BMSCs under oxidative stress,and its mechanism may be through activating of Nrf2/HO-1/NQO1 signaling pathway.

参考文献/References:

[1] Lim M,Wang W,Liang L,et al.Intravenous injection of allogeneic umbilical cord-derived multipotent mesenchymal stromal cells reduces the infarct area and ameliorates cardiac function in a porcine model of acute myocardial infarction[J].Stem Cell Research & Therapy,2018,9(1):129. [2]Huang L,Ma W,Ma Y,et al.Exosomes in mesenchymal stem cells,a new therapeutic strategy for cardiovascular diseases?[J].International Journal of Biological Sciences,2015,11(2):238-245. [3]Yang M,Wen T,Chen H,et al.Knockdown of insulin-like growth factor 1 exerts a protective effect on hypoxic injury of aged BM-MSCs: role of autophagy[J].Stem Cell Research & Therapy,2018,9(1):284. [4]Zhao Y,Li T,Wei X,et al.Mesenchymal stem cell transplantation improves regional cardiac remodeling following ovine infarction[J].STEM CELLS Translational Medicine,2012,1(9):685-695. [5]Suzuki G,Iyer V,Lee T,et al.Autologous mesenchymal stem cells mobilize cKit+and CD133+bone marrow progenitor cells and improve regional function in hibernating myocardium[J].Circulation Research,2011,109(9):1044-1054. [6]Cai M,Shen R,Song L,et al.Bone Marrow Mesenchymal Stem Cells(BM-MSCs)Improve Heart Function in Swine Myocardial Infarction Model through Paracrine Effects[J].Scientific Reports,2016,6(1):28250. [7]Zhao J,Liu X,Kong F,et al.Bone marrow mesenchymal stem cells improve myocardial function in a swine model of acute myocardial infarction[J].Molecular Medicine Reports,2014,10(3):1448-1454. [8]Li Y,Chen J,Chen X G,et al.Human marrow stromal cell therapy for stroke in rat: Neurotrophins and functional recovery[J].Neurology,2002,59(4):514-523. [9]Chen J,Li Y,Wang L,et al.Therapeutic Benefit of Intravenous Administration of Bone Marrow Stromal Cells After Cerebral Ischemia in Rats[J].Stroke,2001,32(4):1005-1011. [10]Cao F,Lin S,Xie X,et al.In vivo visualization of embryonic stem cell survival,proliferation,and migration after cardiac delivery[J].Circulation,2006,113(7):1005-1014. [11]Zhang Z,Li S,Cui M,et al.Rosuvastatin enhances the therapeutic efficacy of adipose-derived mesenchymal stem cells for myocardial infarction via PI3K/Akt and MEK/ERK pathways[J].Basic Research in Cardiology,2013,108(2):333. [12]Cominacini L,Mozzini C,Garbin U,et al.Endoplasmic reticulum stress and Nrf2 signaling in cardiovascular diseases[J].Free Radical Biology and Medicine,2015,88:233-242. [13]唐一锋,石蓓,王冬梅,等.HMGB-1联合MSCs移植对急性心肌梗死大鼠心脏功能影响的实验研究[J].重庆医学,2018,47(12):1583-1588. [14]陈文明,蹇明辉,赵永超,等.Toll样受体4通过PI3K/Akt信号通路调节骨髓间充质干细胞凋亡[J].遵义医学院学报,2017,40(6):617-621. [15]Huang D,Yin L,Liu X,et al.Geraniin protects bone marrow?derived mesenchymal stem cells against hydrogen peroxide?induced cellular oxidative stress in vitro[J].International Journal of Molecular Medicine,2017:47. [16]Vahidinia Z,Azami Tameh A,Nejati M,et al.The protective effect of bone marrow mesenchymal stem cells in a rat model of ischemic stroke via reducing the C-Jun N-terminal kinase expression[J].Pathology - Research and Practice,2019:152519. [17]Discher D E,Mooney D J,Zandstra P W.Growth factors,matrices,and forces combine and control stem cells[J].Science,2009,324(5935):1673-1677. [18]Pittenger M F,Martin B J.Mesenchymal stem cells and their potential as cardiac therapeutics[J].Circulation Research,2004,95(1):9-20. [19]邓宁波,韩腾龙,曾元清,等.人血红素加氧酶1过表达大鼠骨髓间充质干细胞对缺血缺氧环境损伤的耐受能力[J].中国组织工程研究,2017,21(29):4617-4622. [20]陈少强,吴碧莲,王姗姗,等.过表达基质细胞衍生因子1基因促进骨髓间充质干细胞增殖和迁移[J].中国组织工程研究,2018,22(1):32-39. [21]李妍,杜红延,李红卫.慢病毒载体及其在RNA干扰技术中的应用与发展[J].分子诊断与治疗杂志,2013,5(01):55-60. [22]Zhou Y,Wang H,Zhu L,et al.Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line[J].Oncology Reports,2012,29(1):394-400. [23]Chen L,Li K,Liu Q,et al.Protective effects of raspberry on the oxidative damage in HepG2 cells through Keap1/Nrf2-dependent signaling pathway[J].Food and Chemical Toxicology,2019,133:110781. [24]Liu C,Cui Y,Pi F,et al.Torularhodin ameliorates oxidative activity in vitro andd-galactose-induced liver injury via the Nrf2/HO-1 signaling pathway in vivo[J].Journal of Agricultural and Food Chemistry,2019,67(36):10059-10068. [25]Wang T,Sun X,Li A,et al.Botrysphin D attenuates arsenic-induced oxidative stress in human lung epithelial cells via activating Nrf2/ARE signaling pathways[J].Biochemical and Biophysical Research Communications,2019,518(3):526-532. [26]柳源,刘龙中,徐亚沙,等.清胰Ⅱ号对雨蛙素诱导的急性胰腺炎小鼠Nrf2相关基因水平的影响[J].遵义医学院学报,2017,40(1):33-37. [27]聂雨,张斌斌,黄波,等.70味珍珠丸对CCl4致急性肝损伤小鼠的保护作用及机制研究[J].遵义医学院学报,2017,40(4):358-363.

相似文献/References:

[1]刘艳枚,刘小慧,方 宁,等.人VEGF基因腺病毒表达载体修饰大鼠骨髓间充质干细胞[J].遵义医科大学学报,2012,35(01):1.
 Liu yanmei,Liu xiaohui,Fang ning,et al.In vitro modified rat bone marrow mesenchymal stem cells with human vascular endothelial growth factor expressing adenovirus vector[J].Journal of Zunyi Medical University,2012,35(05):1.
[2]唐一锋,马 帅,石 蓓.HMGB1联合MSCs移植对急性心肌梗死大鼠心脏血管生成影响[J].遵义医科大学学报,2014,37(04):378.
 Tang Yifeng,Ma Shuai,Shi Bei.Effects of HMGB1 in combination with MSCs transplantation on angiogenesis after acute myocardial infarction in rats[J].Journal of Zunyi Medical University,2014,37(05):378.
[3]陈文明,蹇明辉,赵永超,等.Toll样受体4通过PI3K/Akt信号通路调节骨髓间充质干细胞凋亡[J].遵义医科大学学报,2017,40(06):617.
 Chen Wenming,Jian Minghui,Zhao Yongchao,et al.Toll-like receptor 4 regulats apoptosis of bone marrow mesenchymal stem cells through PI3K/Akt signaling pathways[J].Journal of Zunyi Medical University,2017,40(05):617.

备注/Memo

备注/Memo:
[基金项目]国家自然科学基金资助项目(NO:81660049); 贵州省科技厅社发处项目(NO:黔科合SY字[2013] 3046)。 [通信作者]石蓓,女,硕士,教授,硕士生导师,研究方向:干细胞与心肌血管生成机制,E-mail:shibei:2147@163.com。
更新日期/Last Update: 2019-10-25