硫酸镍对HaCaT细胞增殖、凋亡及炎症表达的影响  

Effects of Nickel Sulfate on the Proliferation, Apoptosisand Inflammatory Expression of HaCaT Cells

作　　者：朱长华 林立航[1] 苏惠春 程波[2] ZHU Changhua;LIN Lihang;SU Huichun;CHENG Bo(Department of Dermatology,Fujian Medical University Union Hospital,Fuzhou 350001,China;Department of Dermatology,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属协和医院皮肤科,福州350001 [2]福建医科大学附属第一医院皮肤科,福州350005

出　　处：《福建医科大学学报》2023年第4期258-264,共7页Journal of Fujian Medical University

基　　金：北京白求恩基金(B19004CT);福建医科大学协和医院院内经费(2021XH026)。

摘　　要：目的探讨硫酸镍刺激对角质形成细胞增殖、凋亡及炎症表达的影响。方法采用CCK-8法检测不同质量浓度的硫酸镍刺激HaCaT细胞12、24、36、48和72 h后细胞的增殖活力。采用Annexin V-Alexa Fluor 647/PI试剂盒检测不同质量浓度的硫酸镍刺激HaCaT细胞24和48 h后的细胞凋亡率。采用实时荧光定量PCR技术检测不同质量浓度的硫酸镍刺激HaCaT细胞24和48 h后,炎症因子白细胞介素(IL)-1β、IL-6、胸腺间质淋巴生成素(TSLP)和肿瘤坏死因子-α(TNF-α)的mRNA的表达情况。结果与未处理的对照组相比,硫酸镍(100、200和300μg/mL)处理HaCaT细胞12、24、36、48和72 h后,细胞增殖能力受抑制且凋亡细胞比例明显升高。低质量浓度的硫酸镍(50μg/mL)处理时,随着时间的延长,细胞存活率呈现先下降后升高的趋势;高质量浓度的硫酸镍(100、200和300μg/mL)处理时,随着时间的延长,细胞存活率呈现下降的趋势。硫酸镍(50、100、200和300μg/mL)刺激HaCaT细胞24和48 h后,IL-1βmRNA表达降低(P<0.01),IL-6和TSLP的mRNA表达升高(P<0.01),TNF-αmRNA变化不明显(P>0.05)。结论高质量浓度的硫酸镍可抑制HaCaT细胞增殖,促进凋亡。硫酸镍刺激HaCaT细胞,可降低IL-1β的表达,增加TSLP和IL-6的表达。Objective This study investigates the impact of nickel sulfate stimulation on proliferation,apoptosis,and inflammatory expression in keratinocytes.Methods HaCaT cell proliferation was assessed using the Cell Counting Kit-8(CCK-8)assay at 12,24,36,48,and 72 h after exposure to different concentrations of nickel sulfate.Apoptosis was measured with the Annexin V-Alexa Fluor 647/PI kit at 24 and 48 h after exposure to nickel sulfate.Real-time quantitative PCR was utilized to analyze interleukin(IL)-1β,IL-6,thymic stromal lymphopoietin(TSLP),and tumor necrosis factor-alpha(TNF-α)mRNA expression in HaCaT cells at 24 and 48 h after exposure to nickel sulfate.Results Compared to the control group,nickel sulfate stimulation(100,200,and 300μg/mL)suppressed HaCaT cell proliferation and significantly increased apoptosis at 12,24,36,48,and 72 h.At a low concentration of nickel sulfate(50μg/mL),cell viability initially decreased and then increased with prolonged exposure.However,at high concentrations(100,200,and 300μg/mL),cell viability decreased over time.Following 24 and 48 h of nickel sulfate stimulation,IL-1βmRNA decreased significantly(P<0.01),while IL-6 and TSLP mRNA increased significantly(P<0.01).The change in TNF-αmRNA was not significant(P>0.05).Conclusions High concentrations of nickel sulfate suppress HaCaT cell proliferation and induce apoptosis.Nickel sulfate stimulation reduces IL-1βexpression and increases TSLP and IL-6 expression in HaCaT cells.

关 键 词：HACAT细胞 接触性皮炎 硫酸镍 增殖 炎症 

分 类 号：R758.2[医药卫生—皮肤病学与性病学]