[1]唐 彬,等.神经干细胞微囊泡抑制H2O2 诱导DRG 神经元氧化应激损伤机制研究[J].现代检验医学杂志,2022,37(04):159-164.[doi:10.3969/j.issn.1671-7414.2022.04.031]
　TANG Bin,WAN Chang-chun,ZONG Shou-yang,et al.Research on Mechanisms of Neural Stem Cell Microvesicles Inhibit Oxidative Stress Injury in H2O2-Induced DRG Neurons[J].Journal of Modern Laboratory Medicine,2022,37(04):159-164.[doi:10.3969/j.issn.1671-7414.2022.04.031]
点击复制

神经干细胞微囊泡抑制H2O2 诱导DRG 神经元氧化应激损伤机制研究()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年04期
页码:
159-164
栏目:
研究简报·实验技术
出版日期:
2022-07-15

文章信息/Info

Title:
Research on Mechanisms of Neural Stem Cell Microvesicles Inhibit Oxidative Stress Injury in H2O2-Induced DRG Neurons
文章编号:
1671-7414（2022）04-159-06
作者:
唐 彬1;  2; 万长春1; 宗寿洋1; 胡嘉波2
1. 金湖县人民医院，江苏金湖 211600；2. 江苏大学医学院，江苏镇江 212013
Author(s):
TANG Bin1; 2; WAN Chang-chun1; ZONG Shou-yang1; HU Jia-bo2
1.Jinhu County People’s Hospital，Jiangsu Jinhu 211600，China； 2.School of Medicine，Jiangsu University，Jiangsu Zhenjiang 212013，China
关键词:
神经干细胞; 微囊泡; 背根神经节; 氧化应激; 凋亡
分类号:
R446.8
DOI:
10.3969/j.issn.1671-7414.2022.04.031
文献标志码:
A
摘要:
目的 探究神经干细胞微囊泡（neural stem cell microvesicles, NSC-MVs）对H2O2 诱导背根神经节（dorsal rootganglion, DRG）神经元氧化应激损伤的作用及机制。方法 超速离心提取NSC-MVs，并进行电镜和纳米颗粒示踪分析。原代培养大鼠DRG 神经元，β-tubulin Ⅲ荧光染色。建立H2O2 诱导DRG 神经元氧化应激损伤模型，确定作用浓度。经NSC-MVs 预处理，MTT( 四唑盐) 检测神经元活力，流式细胞术检测Annexin Ⅴ和PI，蛋白质印迹检测凋亡相关蛋白cleaved caspase 3，cleaved caspase 9，Bax 和Bcl-2 的表达。结果 NSC-MVs 在透射电镜下呈圆盘状，包膜完整，纳米颗粒示踪显示其粒径为50 ～ 450 nm。MTT 结果显示，与对照组相比，H2O2 组神经元活力明显抑制。当H2O2 浓度为25，50，100 和200μmol/L 时具有显著性差异，细胞活力分别为84.4 %，73.7 %，69.8 % 和49.5 %（F=127.7，P ＜ 0.01）。经100，200 和400 μg/ml 的NSC-MVs 预处理DRG 神经元，细胞活力得到明显提升，分别为51.4 %，67.4 % 和73.5 %（F=49.47，P=0.023）。流式细胞术检测结果显示，与对照组相比，H2O2 组神经元凋亡率显著上升（P ＜ 0.05），NSCMVs预处理组细胞凋亡率明显下降（P ＜ 0.05）。蛋白质印迹结果显示，与H2O2 组相比，NSC-MVs 显著抑制cleavedcaspase3，cleaved caspase 9 和Bax 蛋白表达（均P ＜ 0.05），上调Bcl-2 蛋白表达（P ＜ 0.05）。结论 NSC-MVs 能够抑制H2O2 诱导DRG 神经元氧化应激损伤，发挥神经保护作用。
Abstract:
Objective To investigate the effect and mechanism of neural stem cell microvesicles (NSC-MVs) on oxidative stress injury in H2O2-induced DRG neuron. Methods NSC-MVs were extracted by ultracentrifugation and analyzed by electron microscopy and nanoparticle tracer. Rat dorsal root ganglion (DRG) neurons were cultured and identified by fluorescence staining of β-tubulin Ⅲ . The oxidative stress injury model was established in DRG neurons induced with H2O2 and the concentration was determined. After the pretreatment with NSC-MVs, the cell viability of neurons was detected by MTT, Annexin Ⅴ and PI in DRG neurons were detected by flow cytometry, the apoptosis-related protein expression of cleaved caspase 3, cleaved caspase 9, Bax and Bcl-2 were detected by western blotting. Results The NSC-MVs were disc-shaped with intact envelope under transmission electron microscope, the particle size was 50-450 nm by nanoparticle tracer analysis. MTT assay showed that DRG neuron viability was significantly inhibited in H2O2 group compared with control group. When H2O2 concentration was 25, 50, 100 and 200 μmol/L, the cell viability was 84.4 %, 73.7 %, 69.8 % and 49.5% respectively (F = 127.7, P ＜ 0.01). The viability of DRG neurons pretreated with 100, 200 or 400 μg/ml NSC-MVs was significantly increased to 51.4 %, 67.4 % or 73.5 % (F=49.47，P =0.023). Flow cytometry results showed that apoptotic rate increased remarkably in H2O2 group compared with the control group (P ＜ 0.05), neuronal apoptosis in NSC-MVs pretreatment group was obviously inhibited compared with the H2O2 group (P ＜ 0.05). Western blotting results showed that compared with the H2O2 group, expressions of cleaved caspase 3, cleaved caspase 9 and Bax proteins were down-regulated (all P ＜ 0.05), while Bcl-2 was up-regulated in NSC-MVs-treated group (P ＜ 0.05). Conclusion NSC-MVs can inhibit H2O2-induced DRG neuron oxidative stress damage and play a neuroprotective role.

参考文献/References:

[1] VALKO M, RHODES C J, MONCOL J, et al. Free radicals, metals and antioxidants in oxidative stressinduced cancer[J]. Chemico-Biological Interactions,2006, 160(1): 1-40.
[2] VALKO M, LEIBFRITZ D, MONCOL J, et al. Free radicals and antioxidants in normal physiological functions and human disease[J]. The International Journal of Biochemistry & Cell Biology, 2007, 39(1):44-84.
[3] LI L, ZHU K, LIU Y, et al. Targeting thioredoxin-1 with siRNA exacerbates oxidative stress injury after cerebral ischemia/reperfusion in rats[J]. Neuroscience,2015, 284: 815-823.
[4] 余佳红, 童玉, 叶开, 等. 人神经干细胞微囊泡对谷氨酸诱导PC12 细胞损伤的保护作用[J]. 江苏大学学报(医学版), 2020, 30(4): 302-306. YU Jiahong, TONG Yu, YE Kai, et al. Protective effect of human neural stem cell microvesicles against glutamate-induced cytotoxicity in PC12 cells[J]. Journal of Jiangsu University(Medicine Edition),2020,30(4):302-306.
[5] 张筱东, 谢星星, 李佳, 等. 血清外泌体miR-221 和miR-378 水平检测在胃癌诊断中的应用研究[J]. 现代检验医学杂志, 2020, 35(6):52-55. ZHANG Xiaodong, XIE Xingxing, LI Jia, et al. Application study of serum exosome miR-221 and miR-378 detection in the diagnosis of gastric cancer[J].Journal of Modern Laboratory Medicine,2020,35(6):52-55.
[6] 张智慧, 李想, 黎昱江, 等. 干细胞治疗肺损伤的机遇与挑战[J]. 广东医学, 2021, 42(3): 249-253. ZHANG Zhihui, LI Xiang, LI Yujiang, et al.Opportunities and challenges of stem cell therapy for lung injury [J].Guangdong Medical Journal,2021,42(3):249-253.
[7] TANG Bing, NI Wei, ZHOU Jiqin, et al. Peroxiredoxin 6 secreted by schwann-like cells protects neuron against ischemic stroke in rats via PTEN/PI3K/AKT pathway[J]. Tissue & Cell, 2021, 73: 101635.
[8] 赵芳芳, 郭红, 陈嘉．LncRNA TUG1 在甲状腺癌组织中的表达及其对细胞增殖和迁移的影响[J]. 现代检验医学杂志, 2020, 35(6):42-47. ZHAO Fangfang, GUO Hong, CHEN Jia. Expression of lncRNA TUG1 in thyroid carcinoma tissues and its effect on cell proliferation and migration[J].Journal of Modern Laboratory Medicine,2020,35(6):42-47.
[9] 李策, 张备, 姜从玉, 等. 影响神经干细胞增殖及分化的主要因素[J]. 上海医药, 2021, 42(7):21-25, 43.LI Ce, ZHANG Bei, JIANG Congyu, et al. The factors influencing the proliferation and differentiation of neural stem cells[J].Shanghai Medical & Pharmaceutical Journal,2021,42(7):21-25,43.
[10] VOGEL A, UPADHYA R, SHETTY A K. Neural stem cell derived extracellular vesicles: Attributes and prospects for treating neurodegenerative disorders[J].EBioMedicine,2018,38:273-282.
[11] DE GIOIA R, BIELLA F, CITTERIO G, et al. Neural stem cell transplantation for neurodegenerative diseases[J]. International Journal of Molecular Sciences, 2020, 21(9): 3103.
[12] ROBBINS P D, DORRONSORO A, BOOKER C N. Regulation of chronic inflammatory and immune processes by extracellular vesicles[J]. The Journal of Clinical Investigation, 2016, 126(4): 1173-1180.
[13] YANG Tianzhi, MARTIN P, FOGARTY B, et al.Exosome delivered anticancer drugs across the bloodbrain barrier for brain cancer therapy in danio rerio[J].Pharmaceutical Research, 2015, 32(6): 2003-2014.
[14] PAN Qunwen, KUANG Xiaoli, CAI Shuyun, et al. MiR-132-3p priming enhances the effects of mesenchymal stromal cell-derived exosomes on ameliorating brain ischemic injury[J]. Stem Cell Research & Therapy, 2020, 11(1): 260.
[15] JEYARAM A, JAY S M. Preservation and storage stability of extracellular vesicles for therapeutic applications[J]. The AAPS Journal, 2017, 20(1): 1.
[16] YUAN Ping, DING Lu, CHEN Huili, et al. Neural stem cell-derived exosomes regulate neural stem cell differentiation through miR-9-Hes1 axis[J].Frontiers in Cell and Developmental Biology,2021,9: 601600.
[17] ZHANG Lei, GAO Jianyi, CHEN Tianyan, et al.Microvesicles derived from human embryonic neural stem cells inhibit the apoptosis of HL-1 cardiomyocytes by promoting autophagy and regulating AKT and mTOR via transporting HSP-70[J].Stem Cells International, 2019,2019: 6452684.
[18] ZHONG Dong, CAO Yong, LI Chengjun, et al.Neural stem cell-derived exosomes facilitate spinal cord functional recovery after injury by promoting angiogenesis[J]. Experimental Biology and Medicine(Maywood), 2020, 245(1): 54-65.
[19] 张玮, 唐卉凌, 王筠, 等. 大鼠大脑皮质神经元原代培养不同培养体系及纯化方法的比较研究[J]. 现代生物医学进展, 2010, 10(6): 1043-1046. ZHANG Wei, TANG Huiling, WANG Jun, et al. Comparative study on cortical neuron primary culture system and purification in two groups of rats [J]. Progress in Modern Biomedicine,2010,10(6):1043-1046.
[20] WU Zhiming, WANG Huangen, FANG Sunyang, et al.Roles of endoplasmic reticulum stress and autophagy on H2O2-induced oxidative stress injury in HepG2 cells[J]. Molecular Medicine Reports, 2018, 18(5):4163-4174.
[21] WANG Xueqin, HE Xuemei, DENG Xian, et al. Roles of miR-4463 in H2O2-induced oxidative stress in human umbilical vein endothelial cells[J]. Molecular Medicine Reports, 2017, 16(3): 3242-3252.
[22] 马大亮, 崔红莉, 荣卫江, 等. 柴胡皂苷A 减轻脑缺血再灌注大鼠海马神经元损伤[J]. 中国临床解剖学杂志, 2021, 39(5): 569-574. MA Daliang, CUI Hongli, RONG Weijiang,et al.Saikosaponin A attenuates hippocampal neuron damage in rats with cerebral ischemia-reperfusion [J].Chinese Journal of Clinical Anatomy, 2021,39(5):569-574.

相似文献/References:

[1]张明威a,陈　阳a,王　静a,等.肾细胞癌患者癌组织和血清微囊泡miR-378表达水平的临床应用价值研究[J].现代检验医学杂志,2022,37(02):1.[doi:10.3969/j.issn.1671-7414.2022.02.001]
　ZHANG Ming-weia,CHEN Yanga,WANG Jinga,et al.Clinical Value of miR-378 Expression Levels in Tumor Tissue and Serum Extracellular Vesicles for Renal Cell Carcinoma Patients and Its Function Prediction[J].Journal of Modern Laboratory Medicine,2022,37(04):1.[doi:10.3969/j.issn.1671-7414.2022.02.001]

备注/Memo

备注/Memo:
作者简介：唐彬（1993-），男，硕士研究生，研究方向：神经损伤修复，E-mail: 17826072927@163.com。
更新日期/Last Update: 1900-01-01