Abstract:

Li Xiaoqiong, Wei Zhiping, Liu Yanqun. Department of Dermatology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu, China Corresponding author: Wei Zhiping, Email: xzwzp1961@aliyun.com 【Abstract】 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 groups （both P < 0.05）, but similar between the 25- and 50-μg/L rhPEDF groups （P > 0.05）. Conclusions rhPEDF can inhibit the proliferation of HaCaT cells, and down-regulate the mRNA and protein expressions of IL-6, IL-8 and VEGF.