北京大学学报(医学版) ›› 2017, Vol. 49 ›› Issue (3): 371-375. doi: 10.3969/j.issn.1671-167X.2017.03.001

• 论著 •    下一篇

纳米银对皮肤细胞半通道活性的影响及其在细胞增殖抑制中的作用

杨迪,许珺辉,邓芙蓉△,郭新彪   

  1. (北京大学公共卫生学院劳动卫生与环境卫生学系, 北京100191)
  • 出版日期:2017-06-18 发布日期:2017-06-18
  • 通讯作者: 邓芙蓉 E-mail: lotus321321@126.com
  • 基金资助:
    国家自然科学基金(21077006,21477004)和国家高技术研究发展计划(863计划,2012AA062804)资助

Effects of silver nanoparticle on hemichannel activation and anti-proliferation in HaCaT cells

YANG Di, XU Jun-hui, DENG Fu-rong△, GUO Xin-biao   

  1. (Department of Occupational and Environmental Health, Peking University School of Public Health, Beijing 100191, China)
  • Online:2017-06-18 Published:2017-06-18
  • Contact: DENG Fu-rong E-mail: lotus321321@126.com
  • Supported by:
    Supported by the National Natural Science Foundation of China (21077006,21477004) and the National High Technology Research and Development Program of China (863 program, 2012AA062804)

摘要: 目的:探讨纳米银(silver nanoparticles, AgNPs)对皮肤细胞半通道活性的影响,并进一步研究半通道活性的改变在AgNPs诱导的细胞增殖抑制中的作用。方法:不同浓度(0、0.1、0.2、1.0、5.0、10.0、15.0和20.0 μg/cm2)的AgNPs处理人永生化表皮细胞系(HaCaT)24 h,采用碘化丙啶(propidium iodide, PI)染料排除法测定细胞活性,采用溴化乙锭(ethidium bromide, EB)摄取法测定细胞半通道活性。通过以常规密度(4×104 个/cm2)或低密度(1×104 个/cm2)接种细胞及采用半通道抑制剂甘珀酸(carbenoxolone, CBX)预处理的方法调节细胞半通道活性,采用细胞增殖-毒性检测试剂盒(cell counting kit-8, CCK-8)检测半通道活性受抑制、不同浓度AgNPs作用后的HaCaT细胞的增殖水平。结果:当AgNPs浓度低于10 μg/cm2时,HaCaT细胞的细胞活性未受明显影响。10 μg/cm2的AgNPs作用2、12和24 h可导致细胞半通道活性升高至对照组的116.67%、124.85%和139.53%,与对照组相比差异均具有统计学意义(P<0.01), 且25、50和100 μmol/L CBX预处理可明显抑制10 μg/cm2AgNPs作用24 h 后的细胞的半通道活性(P<0.01)。此外,100 μmol/L CBX预处理可升高低密度和常规密度下10 μg/cm2AgNPs 作用24 h后细胞的增殖水平,其中,低密度下,CBX预处理组与AgNPs单独处理组相比,细胞增殖水平显著升高(P<0.01)。结论:一定浓度的AgNPs可激活HaCaT细胞的半通道,半通道活性升高在AgNPs引发的细胞增殖抑制效应中具有重要作用。

关键词: 纳米银, 半通道, 细胞增殖

Abstract: Objective: To investigate the effect of silver nanoparticles (AgNPs) on hemichannel activity in human skin keratinocytes (HaCaT) and to explore the role of hemichannel in AgNP-induced anti-proliferative effect on HaCaT cells. Methods: HaCaT cells were exposed to 0, 0.1, 0.2, 1.0, 5.0, 10.0, 15.0 and 20.0 μg/cm2 of AgNPs for 24 h and cell viability was assessed by propidium iodide (PI) staining with flow cytometry. Hemichannel activity was examined by ethidium bromide (EB) uptake experiments in cells exposed to AgNPs with and without hemichannel inhibitor carbenoxolone (CBX). Afterward, HaCaT cells were seeded at a low density of 1×104cell /cm2 or a normal density of 4×104cell /cm2 and cultured for 24 h. Cell proliferation was measured by cell counting kit-8 (cck-8) in low-or normal-density cultured cells exposed to AgNPs for 24 h with and without 100 μmol/L CBX. Results: Cell viability showed no significant differences between the control and AgNP-exposed groups with the concentration less than 10 μg/cm2. Exposure to AgNPs increased EB uptake in a time- and dose-dependent manner in HaCaT cells and EB fluoresce density was increased to 116.67%, 124.85% and 139.53% of the control after exposure to 10 μg/cm2 AgNPs for 2 h, 12 h and 24 h. After being treated with 25, 50 and 100 μmol/L CBX, 10 μg/cm2 AgNP-induced increase in EB uptake was significantly reduced in HaCaT cells (P<0.01). The cellular proliferation rate was increased inlow-and normal-density cultured cells after AgNPs exposure with 100 μmol/L CBX. After being treated with 100 μmol/L CBX, 10 μg/cm2 AgNP-induced anti-proliferation in low-density cultured cells was significantly restrained (P<0.01). Conclusion: AgNPs could enhance hemichannel activity of HaCaT cells. Hemichannel activation was involved in AgNP-induced anti-proliferative effect.

Key words: Silver nanoparticles, Hemichannel, Cell proliferation

中图分类号: 

  •  
[1] 张瑶,郭金鑫,战世佳,洪恩宇,杨慧,贾安娜,常艳,郭永丽,张璇. 富含半胱氨酸和甘氨酸蛋白2在神经母细胞瘤恶性进展中的功能和机制[J]. 北京大学学报(医学版), 2024, 56(3): 495-504.
[2] 刘媛,原婉琼,李婷,王平章,吕平,吴利新,阮国瑞,韩文玲,莫晓宁. 敲减CMTM3增加急性B淋巴细胞白血病细胞对伊马替尼敏感性[J]. 北京大学学报(医学版), 2022, 54(6): 1238-1243.
[3] 谢静,赵玉鸣,饶南荃,汪晓彤,方滕姣子,李晓霞,翟越,李静芝,葛立宏,王媛媛. 3种口腔颌面部来源的间充质干细胞成血管内皮分化潜能的比较研究[J]. 北京大学学报(医学版), 2019, 51(5): 900-906.
[4] 王子成,程立,吕同德,苏黎,林健,周利群. 炎症因子预处理的脂肪干细胞可明显抑制外周血单个核细胞增殖[J]. 北京大学学报(医学版), 2018, 50(4): 590-594.
[5] 唐旭,赵卫红,宋琴琴,殷华奇,杜依青,盛正祚,王强,张晓威,李清,刘士军,徐涛. SOX10对前列腺癌细胞增殖及侵袭的影响[J]. 北京大学学报(医学版), 2018, 50(4): 602-606.
[6] 陈玮, 胡凡磊, 刘洪江, 徐丽玲, 李英妮, 栗占国. 类风湿关节炎患者髓系来源的抑制细胞促进自身B细胞增殖[J]. 北京大学学报(医学版), 2017, 49(5): 819-823.
[7] 蔡燚,郭浩,李汉忠,王文达,张玉石. 结节性硬化症细胞株TSC2-/- MEFs和正常细胞株TSC2+/+ MEFs微小RNA表达谱的差异分析[J]. 北京大学学报(医学版), 2017, 49(4): 580-584.
[8] 高翔,陈香梅,张婷,张静,陈茉,郭正阳,石岩岩,鲁凤民,丁士刚. 巨噬细胞加帽蛋白与胃癌细胞增殖及迁移能力的关系[J]. 北京大学学报(医学版), 2017, 49(3): 489-494.
[9] 司马梓涵,洪瑛瑛,李铁军. PTCH1基因突变对牙源性角化囊性瘤上皮细胞增殖的影响[J]. 北京大学学报(医学版), 2017, 49(3): 522-526.
[10] 隋华欣, 吕培军, 王宇光, 王勇, 孙玉春. 低能量激光照射对人脂肪基质细胞增殖分化的影响[J]. 北京大学学报(医学版), 2017, 49(2): 337-343.
[11] 李静文,殷晓晖,栾庆先. 不同代次诱导多能干细胞增殖及牙周定向分化的能力[J]. 北京大学学报(医学版), 2017, 49(1): 16-024.
[12] 胡佳,邹晓英,庄姮,高学军. 根管封闭剂对牙周膜细胞生物相容性的影响[J]. 北京大学学报(医学版), 2016, 48(5): 871-877.
[13] 凌龙,赵玉鸣,葛立宏. 不同炎症状态下犬年轻恒牙牙髓干细胞增殖及成骨分化能力的改变[J]. 北京大学学报(医学版), 2016, 48(5): 878-883.
[14] 莫珩,高承志,王少杰,李玫,董建强,郁卫东. 口腔鳞癌中CD24的表达、作用和治疗靶点价值[J]. 北京大学学报(医学版), 2016, 48(1): 16-22.
[15] 杨颖婷, 卫彦, 夏斌, 王珍高. 不同纤维直径聚吡咯通电后对细胞和细菌黏附增殖的影响[J]. 北京大学学报(医学版), 2016, 48(1): 63-70.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!