摘要
目的 通过阿维A作用于缺氧培养的HaCaT细胞，初步探讨阿维A治疗银屑病的可能机制。方法 将浓度为10-5、10-6、10-7、10-8 mol/L的阿维A分别作用于HaCaT细胞，用CCK-8法检测缺氧培养12、24、36 h对HaCaT细胞体外增殖的影响。反转录PCR和 Western印迹检测不同浓度阿维A对缺氧培养24 h的HaCaT细胞HIF-1α、血管内皮生长因子（VEGF）蛋白及mRNA表达的影响。结果 阿维A 10-8、10-7、10-6、10-5 mol/L作用24 h，HaCaT细胞体外增殖抑制率分别为（13.31 ± 1.15）%、（21.86 ± 5.31）%、（32.05 ± 2.99）%、（37.28 ± 3.21）%，且随着缺氧培养时间延长和阿维A浓度增加，对HaCaT细胞体外增殖的抑制作用逐渐增强。当阿维A浓度为10-5 mol/L时，HIF-1α蛋白的表达由1.196 ± 0.088降至0.319 ± 0.180（P＜0.05），而HIF-1α mRNA表达水平无明显下降；VEGF mRNA的表达由1.108 ± 0.073 降至0.442 ± 0.090（P＜0.05）， VEGF蛋白的表达则由1.174 ± 0.186降至 0.216 ± 0.066（P＜0.05）。结论 阿维A可抑制缺氧培养的HaCaT细胞体外增殖，并在蛋白水平下调HIF-1α及VEGF的表达，在mRNA水平下调VEGF的表达。
Objective To evaluate effects of acitretin on HaCaT cells cultured in hypoxic condition, and to preliminarily explore the possible therapeutic mechanisms of acitretin in psoriasis. Methods HaCaT cells were divided into several groups to be cultured in hypoxic condition with the presence of acitretin at concentrations of 10-5, 10-6, 10-7 and 10-8 mol/L respectively, with cells treated with dimethyl sulfoxide （DMSO） as DMSO control group and those receiving no treatment as blank control group. Cellular proliferative activity was evaluated by CCK-8 assay after 12-, 24- and 36-hour hypoxic culture in vitro. The mRNA and protein s of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF） were determined by reverse transcription （RT）-PCR and Western-blot analysis, respectively, after 24-hour hypoxic culture. Results After 24-hour hypoxic culture, the cellular proliferation rate was inhibited by 13.31% ± 1.15%, 21.86% ± 5.31%, 32.05% ± 2.99% and 37.28% ± 3.21% in the 10-8-, 10-7-, 10-6- and 10-5-mol/L acitretin groups respectively. With the increase of culture duration and acitretin concentrations, the degree of inhibition on cellular proliferation increased gradually. Compared with the blank control group, the 10-5-mol/L acitretin group showed significantly decreased protein of HIF-1α （0.319 ± 0.180 vs. 1.196 ± 0.088, P 〈 0.05）, as well as decreased mRNA and protein s of VEGF （mRNA： 0.442 ± 0.090 vs. 1.108 ± 0.073; protein： 0.216 ± 0.066 vs. 1.174 ± 0.186; both P 〈 0.05）. However, no significant difference was found in the mRNA of HIF-1α between the 10-5-mol/L acitretin group and blank control group. Conclusion Acitretin can suppress the in vitro proliferation of HaCaT cells cultured in hypoxic condition, and down-regulate the s of HIF-1α and VEGF proteins as well as VEGF mRNA.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2016年第9期616-620,共5页
Chinese Journal of Dermatology
