重组人色素上皮衍生因子对 HaCaT 细胞体外增殖和白细胞介素6、8及血管内皮生长因子表达的影响  

Effects of recombinant human pigment epithelium derived factor on in vitro proliferation of and expressions of interleukin-6,-8 and vascular endothelial growth factor in cultured human HaCaT keratinocytes

作　　者：李小琼[1] 魏志平[1] 刘彦群[1] 

机构地区：[1]徐州医学院附属医院皮肤科,江苏221002

出　　处：《中华皮肤科杂志》2015年第8期578-580,共3页Chinese Journal of Dermatology

摘　　要：目的：探讨重组人色素上皮衍生因子（rhPEDF）对 HaCaT 细胞体外增殖和白细胞介素6（IL-6）、IL-8及血管内皮生长因子（VEGF）表达的影响。方法 HaCaT 细胞分4组进行不同处理，分别为100μg/L rhPEDF 组、50μg/L rhPEDF 组、25μg/L rhPEDF 组及对照组（只加 RPMI 1640培养液）。采用 CCK8法检测不同浓度 rhPEDF 作用于 HaCaT 细胞24、48、72 h 后对细胞体外增殖的影响。RT-PCR 和 Western 印迹法检测rhPEDF 对 HaCaT 细胞 IL-6、IL-8、VEGF mRNA 及其蛋白表达的影响。统计分析方法采用两因素方差分析、单因素方差分析、SNK-q 检验以及 Pearson 相关分析。结果25、50、100μg/L 的 rhPEDF 作用24、48、72 h 后对HaCaT 细胞体外增殖均有不同程度的抑制作用。随时间延长和 rhPEDF 浓度的增加，rhPEDF 对 HaCaT 细胞体外增殖的抑制作用均逐渐增强（F =1115、329.9，均 P 〈0.001）。与对照组相比，不同浓度（100、50、25μg/L） rhPEDF 作用于 HaCaT 细胞48 h 后，VEGF、IL-6、IL-8 mRNA 及蛋白的表达均下降，差异均有统计学意义（P 〈0.05）。随 rhPEDF 浓度的增加，各浓度 rhPEDF 组 VEGF mRNA、IL-6和 IL-8蛋白表达均出现下调，与对照组比较，差异均有统计学意义（P 〈0.05）；100μg/L rhPEDF 组的 IL-6、IL-8 mRNA 表达明显低于25μg/L rhPEDF 组（P 〈0.05）；100μg/L rhPEDF 组 VEGF 蛋白表达分别低于50、25μg/L rhPEDF 组（P 〈0.05），而50与25μg/L rhPEDF 组间差异无统计学意义（P 〉0.05）。结论 rhPEDF 可抑制 HaCaT 细胞体外增殖，并可在mRNA 及蛋白水平下调 IL-6、IL-8、VEGF 的表达。Objective To investigate the effects of recombinant human pigment epithelium derived factor （rhPEDF）on in vitro proliferation of and expressions of interleukin 6（IL-6）, IL-8 and vascular endothelial growth factor （VEGF）in cultured human HaCaT keratinocytes. Methods Some cultured HaCaT cells were treated with rhPEDF at various concentrations（25, 50, 100 μg/L）for different durations, and some treated with RPMI 1640 medium only served as the control group. Cell counting kit-8（CCK8）assay was performed to evaluate cell proliferation after 24-, 48- and 72-hour treatment, reverse transcription （RT）-PCR to measure the mRNA expressions of IL-6, IL-8 and VEGF in HaCaT cells after 24-hour treatment, and Western blot to detect the protein expressions of IL-6, IL-8 and VEGF in HaCaT cells after 48-hour treatment. Statistical analysis was carried out by two- and one-way analysis of variance, Student-Newman-Keuls（SNK）-q test and Pearson correlation analysis. Results After treatment with rhPEDF of 25-100 μg/L for 24 - 72 hours, the proliferation of HaCaT cells was significantly inhibited to different extents compared with the control group（all P 〈 0.05）, and the inhibition rate significantly increased with the increase in treatment duration and concentrations of rhPEDF（F = 1115, 329.9, respectively, both P 〈 0.001）. Moreover, there was a significant decrease in the expressions of IL-6, IL-8 and VEGF mRNAs（at 24 hours）and proteins（at 48 hours）in HaCaT cells after treatment with rhPEDF of 25 - 100 μg/L compared with the control group（all P 〈 0.05）. The expression levels of VEGF mRNA as well as IL-6 and IL-8 proteins all significantly decreased with the increase of rhPEDF concentrations （all P 〈 0.05）. The mRNA expressions of IL-6 and IL-8 were significantly lower in the 100-μg/L rhPEDF group than in the 25-μg/L rhPEDF group （both P 〈 0.05）, and the protein expression of VEGF was significantly weaker in the 100-μg/L rhPEDF group than in 25-and 50-μg/L rhPEDF g

关 键 词：角蛋白细胞 白细胞介素8 白细胞介素6 血管内皮生长因子类 银屑病 色素上皮衍生因 子 HaCaT细胞 

分 类 号：R751[医药卫生—皮肤病学与性病学]