中华皮肤科杂志 ›› 2017, Vol. 50 ›› Issue (6): 431-435.

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葫芦素I对HaCaT细胞体外增殖及对角蛋白17、STAT3、VEGF表达的影响

姚秋楠1,魏志平2,刘彦群3   

  1. 1. 徐州医科大学附属医院
    2. 江苏省徐州市徐州医学院附属医院皮肤科
    3. 徐州医学院附属医院皮肤科
  • 收稿日期:2016-09-06 修回日期:2017-02-27 发布日期:2017-05-31
  • 通讯作者: 魏志平 E-mail:xzwzp1961@aliyun.com

Effects of cucurbitacin I on in vitro proliferation of HaCaT cells and the of keratin 17, signal transducer and activator of transcription 3 and vascular endothelial growth factor

  • Received:2016-09-06 Revised:2017-02-27 Published:2017-05-31

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摘要： 目的 探讨葫芦素I对HaCaT细胞体外增殖及角蛋白17（K17）、信号转导及转录激活子3（STAT3）、血管内皮生长因子（VEGF）表达的影响。方法 采用不同浓度葫芦素I（0.0125、0.025、0.05、0.1 μmol/L）、含与0.1 μmol/L葫芦素I等体积DMSO的DMEM培养液（溶媒组）、DMEM培养液（阴性对照组）、10 nmol/L卡泊三醇（阳性对照组）分别作用于HaCaT细胞。采用CCK8法检测葫芦素I作用12、24、36 h后对HaCaT细胞体外增殖的影响，RT?PCR法检测葫芦素I作用24 h对HaCaT细胞K17和VEGF mRNA表达的影响，Western印迹法检测葫芦素I作用24 h对HaCaT细胞K17、STAT3、磷酸化STAT3（P?STAT3）和VEGF蛋白表达的影响。结果 0.0125 μmol/L葫芦素I作用12 h即开始表现出对HaCaT细胞体外增殖的抑制作用，当葫芦素I浓度增加到0.1 μmol/L时，24 h和36 h的抑制率分别为43.00% ± 2.11%和48.98% ± 2.27%。与阴性对照组相比，各浓度葫芦素I组对HaCaT细胞体外增殖均有抑制作用（均P < 0.05），且该抑制作用随葫芦素I浓度的增加及作用时间的延长逐渐增强。不同浓度葫芦素I作用于HaCaT细胞24 h后，K17 mRNA及其蛋白、P?STAT3蛋白、VEGF mRNA及其蛋白表达量均随葫芦素I浓度的增加而降低，差异有统计学意义（均P < 0.05）。结论 葫芦素I可抑制HaCaT细胞的体外增殖，并可在mRNA水平下调K17、VEGF的表达，在蛋白水平下调K17、 P?STAT3、VEGF的表达。

Abstract: Yao Qiunan,Wei Zhiping, Liu Yanqun Department of Dermatology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu, China （The current affiliation of the third author was Jiangsu Medical Association） Corresponding author: Wei Zhiping, Email: xzwzp1961@aliyun.com 【Abstract】 Objective To evaluate effects of cucurbitacin I on in vitro proliferation of HaCaT cells and the of keratin 17 （K17）, signal transducer and activator of transcription 3 （STAT3） and vascular endothelial growth factor （VEGF） in cultured HaCaT cells. Methods In vitro cultured HaCaT cells were divided into 6 groups to be treated with cucurbitacin I at different concentrations of 0.0125, 0.025, 0.05 and 0.1 μmol/L （0.0125, 0.025, 0.05 and 0.1 μmol/L cucurbitacin I groups）, DMEM containing the same volume of DMSO as 0.1 μmol/L cucurbitacin I （DMSO group）, DMEM （negative control group） and 10 nmol/L calcipotriol （positive control group）, respectively. Cell counting kit?8 （CCK8） assay was performed to assess in vitro cellular proliferative activity after 12?, 24?, 36?hour treatment, reverse transcription （RT）?PCR to measure the mRNA of K17 and VEGF in HaCaT cells after 24?hour treatment, and Western blot analysis to determine the protein of K17, STAT3, phosphorylated?STAT3 （p?STAT3） and VEGF after 24?hour treatment. Statistical analysis was carried out by one?way analysis of variance （ANOVA）, repeated measures ANOVA, Student?Newman?Keuls （SNK）?q test and Pearson correlation analysis. Results The proliferative activity of HaCaT cells started to decrease after 12?hour treatment with cucurbitacin I at the concentration of 0.0125 μmol/L. When the concentration of cucurbitacin I increased up to 0.1 μmol/L, the cell proliferation rates were inhibited by 43.00% ± 2.11% and 48.98% ± 2.27% after 24? and 36?hour treatment respectively. Compared with the negative control group, cucurbitacin I at different concentrations all could inhibit in vitro proliferation of HaCaT cells （all P < 0.05）, and the inhibitory effects increased gradually with the increase of cucurbitacin I concentration and treatment duration. After 24?hour treatment, the mRNA of K17 and VEGF and the protein of K17, VEGF and P?STAT3 were significantly decreased along with the increase of concentration of cucurbitacin I （all P < 0.05）. Conclusion Cucurbitacin I can inhibit in vitro proliferation of HaCaT cells, and down?regulate the mRNA of K17 and VEGF and protein of K17, VEGF and P?STAT3.

中图分类号: 

  • R75

引用本文

姚秋楠 魏志平 刘彦群. 葫芦素I对HaCaT细胞体外增殖及对角蛋白17、STAT3、VEGF表达的影响[J]. 中华皮肤科杂志, 2017,50(6):431-435. doi: