棕榈酸对HaCaT细胞增殖及产生炎症因子的影响  

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

作　　者：张倩[1] 周炳荣[1] 骆丹[1] 方晓波[1] 尹慧彬[1] 郭泽[1] 吴唯[1] 

机构地区：[1]南京医科大学第一附属医院皮肤科,210029

出　　处：《中华皮肤科杂志》2013年第9期656-660,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81171518、81000700）

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、200μmol／L）刺激HaCaT细胞3～24h，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、150μmol／L）刺激HaCaT细胞24h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）-κBp65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自介素6（IL-6）分泌量，实时PCR法检测过氧化物酶增殖体激活受体（PPARa）mRNA及IL-6mRNA的表达，免疫印迹法检测PPARa、总蛋白NF—κBp65及核蛋白NF—κBp65表达量。用GraphPadPrism5,0软件进行单因素方差分析。结果与空白对照组相比，50～175μmoVL棕榈酸均可刺激HaCaT细胞增殖（均P〈0．05）。75、100、125、150μmol／L棕榈酸可剂量依赖性增强HaCaT细胞NF—κBp65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）。PPARamRNA及其蛋白产物、IL～6mRNA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．（）1）。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PPARa表达量，在激活和促进相关炎症因子表达中起到一定作用。Objective To estimate the influence of palmitic acid （PA） on the proliferation of and production of inflammatory mediators by a human keratinoeyte 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, 150μmol/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 transloeation 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-aetivated 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 150μmol/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 HaCaY 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, respective

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 

分 类 号：R758.733[医药卫生—皮肤病学与性病学]