中性粒细胞趋化因子3可影响神经干细胞的存活和增殖 |
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作者姓名: | 孙耐 王大力 张君 曹金明 仇福成 刘惠苗 李冬 顾平 |
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作者单位: | 华北理工大学附属医院,河北省唐山市 063000; 河北医科大学第一医院神经内二科,脑老化与认知神经科学实验室,河北省石家庄市 050031;河北医科大学第一医院细胞治疗实验室,河北省石家庄市 050031 |
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基金项目: | 河北省2017年度医学科学研究重点课题计划项目(20170062),项目负责人:顾平;河北医科大学第一医院“星火”青年科研项目(XH201702),项目负责人:张君~~ |
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摘 要: |
文题释义:
神经干细胞:广泛存在于胚胎及成年中枢神经系统内,具有自我复制和分化为神经元、星形胶质细胞和少突胶质细胞的潜能,对脑损伤有较大的修复作用,为临床治疗神经系统疾病带来了新的希望。
中性粒细胞趋化因子3:是含ELR的CXC类趋化因子,主要由激活的单核细胞、内皮细胞、成纤维细胞产生,其在炎症反应中可以诱导白细胞的趋化作用,与受体CXCR2有高度的亲和力,中性粒细胞趋化因子3与其受体CXCR2结合可能产生抗凋亡、促进少突胶质细胞增殖和抑制其迁移的功能。
背景:课题组前期研究首次发现骨髓基质细胞可以分泌中性粒细胞趋化因子3,且在骨髓基质细胞调控神经干细胞分化为神经元的过程中中性粒细胞趋化因子3的表达上调1.5倍,提示中性粒细胞趋化因子3可能具有促进神经干细胞增殖与分化的作用。
目的:观察中性粒细胞趋化因子3对新生大鼠海马来源神经干细胞存活和增殖的影响。
方法:体外分离培养新生SD大鼠海马神经干细胞,将培养3代的神经干细胞分为对照组和中性粒细胞趋化因子3组,通过MTS法检测细胞存活率,细胞生长曲线及活/死细胞染色观察细胞存活及增殖情况,通过免疫荧光染色法及Real-time PCR检测神经干细胞中Nestin的表达来观察神经干细胞增殖情况。
结果与结论:①随着中性粒细胞趋化因子3质量浓度从1
μg/L增加至20 μg/L,神经干细胞存活率逐渐增加,当质量浓度为10 μg/L时,神经干细胞存活率最高且细胞活力最好,再增至20 μg/L时,神经干细胞存活率无显著增加;②中性粒细胞趋化因子3组Nestin染色阳性率明显高于对照组(P <
0.05);③中性粒细胞趋化因子3组的Nestin
mRNA表达量明显高于对照组(P < 0.05);④结果表明,中性粒细胞趋化因子3对海马来源神经干细胞具有促进存活和增殖的作用。
ORCID: 0000-0002-4733-3668(顾平)
中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程
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关 键 词: | 中性粒细胞趋化因子3 海马 神经干细胞 细胞存活 细胞增殖 巢蛋白 |
收稿时间: | 2019-03-13 |
Effects of cytokine-induced neutrophil chemoattractant 3 on the survival and proliferation of neural stem cells |
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Authors: | Sun Nai Wang Dali Zhang Jun Cao Jinming Qiu Fucheng Liu Huimiao Li Dong Gu Ping |
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Institution: | Affiliated
Hospital of North China University of Science and Technology, Tangshan 063000,
Hebei Province, China; Department of Neurology, First Hospital of
Hebei Medical University, Neuroscience Laboratory for Brain Aging and
Cognition, Shijiazhuang 050031, Hebei Province, China; Laboratory
of Cell Therapy, First Hospital of Hebei Medical University, Shijiazhuang
050031, Hebei Province, China |
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Abstract: | BACKGROUND:Our previous study was the first to find that bone marrow stromal cells could secrete neutrophil chemokine-3.Moreover,the expression of cytokine-induced neutrophil chemoattractant 3(CINC-3)could be up-regulated by 1.5 times during the differentiation of neural stem cells into neurons regulated by bone marrow stromal cells.These results suggest that CINC-3 may promote the proliferation and differentiation of neural stem cells.OBJECTIVE:To observe the effect of CINC-3 on survival and proliferation of hippocampal neural stem cells in neonatal rats.METHODS:Hippocampal neural stem cells from neonatal Sprague-Dawley rats were isolated and cultured in vitro,and passage 3 neural stem cells were divided into control and CINC-3 groups.The survival rate of the cells was measured by MTS method,cell survival and proliferation were observed using cell growth curve and live/dead cell staining,and the expression of Nestin in neural stem cells was detected by real-time PCR and immunofluorescence staining for observation of neural stem cell proliferation.RESULTS AND CONCLUSION:(1)When the concentration of CINC-3 increased from 1 to 20μg/L,the survival rate of neural stem cells increased gradually.When the concentration of CINC-3 was 10μg/L,the survival rate of neural stem cells was the highest and the cell viability was the best(P<0.05).When the concentration of CINC-3 further increased to 20μg/L,there was no significant difference in the survival rate of neural stem cells.(2)The positive rate of Nestin in the CINC-3 group was significantly higher than that in the control group(P<0.05).(3)The expression of Nestin mRNA in the CINC-3 group was significantly higher than that in the control group(P<0.05).These findings indicate CINC-3 can promote the survival and proliferation of hippocampal neural stem cells. |
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Keywords: | neutrophil chemoattractant 3 hippocampus neural stem cells survival proliferation nestin |
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