中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (9): 656-660.

• 论著 • 上一篇    下一篇

棕榈酸对HaCaT细胞增殖及产生炎症因子的影响

张倩1,2,周炳荣3,骆丹4,方晓波5,6,尹慧斌7,郭泽8,吴唯2   

  1. 1. 南京市鼓楼医院病理科
    2. 南京医科大学第一临床医学院
    3. 南京医科大学第一附属医院
    4. 南京市南京医科大学附属第一医院皮肤科
    5.
    6. 南京医科大学附属第一医院
    7. 江苏省人民医院
    8. 南京医科大学第一附属医院江苏省人民医院
  • 收稿日期:2012-10-09 修回日期:2013-05-16 出版日期:2013-09-15 发布日期:2013-09-01
  • 通讯作者: 骆丹 E-mail:daniluo2005@163.com
  • 基金资助:
    国家自然科学基金;国家自然科学基金

Influence of palmitic acid on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT

  • Received:2012-10-09 Revised:2013-05-16 Online:2013-09-15 Published:2013-09-01
  • Supported by:
    National Natural Science Foundation of China; National Natural Science Foundation of China

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摘要： 【摘要】 目的 探讨棕榈酸对HaCaT细胞增殖及产生炎症因子的影响。方法 将不同浓度棕榈酸（0、25、50、75、100、125、150、175、200 μmol/L）刺激HaCaT细胞3 ~ 24 h，用细胞计数试剂盒CCK8检测HaCaT细胞增殖情况。选取一定浓度棕榈酸（0、75、100、125、150 μmol/L）刺激HaCaT细胞24 h后，分别用免疫组化荧光染色法在共聚焦显微镜下观察核因子（NF）-κB p65细胞核转位情况。酶联免疫吸附试验检测细胞培养上清液中白介素6（IL-6）分泌量，实时PCR法检测过氧化物酶增殖体激活受体α（PPARα）mRNA及IL-6 mRNA的表达，免疫印迹法检测PPARα、总蛋白NF-κB p65及核蛋白NF-κB p65表达量。用GraphPad Prism 5.0 软件进行单因素方差分析。结果 与空白对照组相比，50 ~ 175 μmol/L棕榈酸均可刺激HaCaT细胞增殖（均P < 0.05）。75、100、125、150 μmol/L棕榈酸可剂量依赖性增强HaCaT细胞NF-κB p65向细胞核内转位，各浓度组核蛋白的相对表达量分别为0.4536 ± 0.0173、0.5184 ± 0.0206、0.5333 ± 0.0231、0.6160 ± 0.0297，与空白对照组相比，差异有统计学意义（均P < 0.01）。PPARα mRNA及其蛋白产物、IL-6 mRNA及分泌量呈剂量依赖性增加，各浓度组IL-6分泌量分别为（31.5677 ± 0.2268）、（32.3773 ± 0.4156）、（32.9837 ± 0.0029）、（33.6890 ± 0.0936） ng/L，与空白对照组相比，差异有统计学意义（P < 0.05或0.01）。结论 棕榈酸可促进HaCaT细胞增殖，并在一定浓度内剂量依赖性增强NF-κB核转移、IL-6及PPARα表达量，在激活和促进相关炎症因子表达中起到一定作用。 【关键词】 棕榈酸； 角蛋白细胞； 细胞增殖； NF-κB； 白细胞介素6； 过氧化物酶增殖体激活受体α

关键词: 角蛋白细胞, 棕榈酸, 细胞增殖, 过氧化物酶增殖体激活受体α, 白细胞介素6, NF-κB

Abstract: ZHANG Qian, ZHOU Bing-rong, LUO Dan, FANG Xiao-bo, YIN Hui-bin, GUO Ze, WU Wei. Department of Dermatology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China Corresponding author: LUO Dan, Email: daniluo2013@njmu.edu.cn 【Abstract】 Objective To estimate the influence of palmitic acid （PA） on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT. Methods Cultured HaCaT cells were treated with PA of eight concentrations （0 － 200 μmol/L） for 3－24 hours followed by the evaluation of cell proliferation by using the cell counting kit-8. According to the proliferation assay, four concentrations （75, 100, 125, 150 μmol/L） of PA were selected and used to treat HaCaT cells for 24 hours, then, fluorescence-based immunohistochemical staining was performed to observe the nuclear translocation of nuclear factor （NF）-κB p65, enzyme linked immunosorbent assay （ELISA） to determine the level of interleukin （IL）-6 in the supernatant of culture medium, real-time PCR to detect the mRNA expressions of peroxisome proliferator-activated receptor α （PPARα） and IL-6, and Western blot to quantify the protein expressions of PPARα as well as total and nuclear NF-κB p65. Those HaCaT cells receiving no treatment served as the control group. Statistical analysis was carried out by one-factor analysis of variance using the GraphPad Prism 5.0 software. Results The HaCaT cells treated with PA of 50 － 175 μmol/L showed accelerated proliferation compared with the control HaCaT cells （all P < 0.05）. PA from 75 to 150 μmol/L enhanced the nuclear translocation of NF-κB p65, mRNA and protein expressions of PPARα, as well as the mRNA expression and supernatant level of IL-6 in a dose-dependent manner. The relative expression level of nuclear NF-κB p65 protein was 0.4536 ± 0.0173, 0.5184 ± 0.0206, 0.5333 ± 0.0231, 0.6160 ± 0.0297, and the supernatant level of IL-6 was （31.5677 ± 0.2268）, （32.3773 ± 0.4156）, （32.9837 ± 0.0029） and （33.6890 ± 0.0936） ng/L, in HaCaT cells treated with PA of 75, 100, 125 and 150 μmol/L, respectively, compared to 0.3237 ± 0.0114 （all P < 0.01） and （30.4577 ± 0.5131） ng/L （all P < 0.01） in the control HaCaT cells, respectively. Conclusions PA can accelerate the proliferation of HaCaT cells, enhance NF-κB nuclear transfer, PPARα expression and IL-6 secretion in a dose-dependent manner within a certain concentration range, and may exert a promoting role in the activation and expression of some inflammatory factors. 【Key words】 Palmitic acid; Keratinocytes; Cell proliferation; NF-kappa B; Interleukin-6; PPARα

Key words: Palmitic acid