新型维A酸ECPIRM及全反式维A酸对白细胞介素17刺激HaCaT细胞白细胞介素1β表达的影响  

Effects of a novel retinoic acid ECPIRM and all-trans.retinoic acid on interleukin-1β expression in interleukin- 17-stimulated HaCaT cells

作　　者：杨林芳[1] 曹玉萍[1] 张孟丽[1] 马鹏程[1] 李玲珺[1] 魏峻[1] 陶蕾 刘维达[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所药物研究室,南京210042

出　　处：《中华皮肤科杂志》2015年第11期797-800,共4页Chinese Journal of Dermatology

摘　　要：目的 探讨新型维A酸ECPIRM及全反式维A酸（ATRA）对白细胞介素17（IL-17）刺激人表皮角质形成细胞株（HaCaT细胞）IL-1β表达的影响。 方法 MTT法检测不同浓度ECPIRM及ATRA与IL-17共同作用于HaCaT细胞后对其细胞增殖活力的影响；IL-17（20 μg/L）刺激HaCaT细胞或与ECPIRM（80 μmol/L）、ATRA（5 μmol/L）共培养24 h，收集细胞培养上清液并提取总RNA，分别进行酶联免疫吸附实验（ELISA）和荧光定量PCR，检测IL-1β蛋白及IL-1β mRNA水平的变化。数据处理采用单因素方差分析，组间比较采用LSD法。 结果 低浓度ECPIRM可促进HaCaT细胞增殖，随着药物浓度和时间的增加，对HaCaT细胞生长出现抑制现象，并呈浓度及时间依赖性。ATRA抑制HaCaT细胞生长呈浓度及时间依赖性。IL-17刺激HaCaT细胞后细胞培养上清液中IL-1β蛋白及IL-1β mRNA水平（20.312 ± 2.053 ng/L、4.05 ± 0.47）与对照组（11.427 ± 0.929 ng/L、1.00 ± 0.03）相比明显升高，差异有统计学意义（P 〈 0.05）；ECPIRM作用后IL-1β蛋白及IL-1β mRNA水平（12.470 ± 1.897 ng/L、0.82 ± 0.12）与IL-17刺激组（20.312 ± 2.053 ng/L、4.05 ± 0.47）相比降低，差异有统计学意义（P 〈 0.05）；ATRA作用后IL-1β蛋白及IL-1β mRNA水平（12.694 ± 1.478 ng/L、0.87 ± 0.16）与IL-17刺激组（20.312 ± 2.053 ng/L、4.05 ± 0.47）相比明显降低，差异有统计学意义（P 〈 0.05），同时ECPIRM处理组、ATRA处理组与溶媒对照组比较差异无统计学意义。 结论 IL-17可促进HaCaT细胞分泌IL-1β，ECPIRM及ATRA对其诱导的HaCaT细胞IL-1β的表达具有明显的抑制作用。Objective To explore the effects of a novel retinoic acid ECPIRM and all-trans-retinoic acid （ATRA） on interleukin （IL）-1β expression in IL-17-stimulated human HaCaT keratinocytes. Methods Cultured HaCaT cells were classified into several groups： ECPIRM groups treated with 20 - 320 μmol/L ECPIRM for 24-72 hours, ATRA groups treated with 1 - 100 μmol/L ATRA for 24 - 72 hours, IL-17 group treated with 20 μg/L IL-17 for 24 hours, IL-17 ＋ ECPIRM group cotreated with 20 μg/L IL-17 and 80 μmol/L ECPIRM for 24 hours, IL-17 ＋ ATRA group cotreated with 20 μg/L IL-17 and 5 μmol/L ATRA for 24 hours, control group treated with the solvent solution of IL-17. Then, methyl thiazolyl tetrazolium （MTT） assay was performed to evaluate cellular proliferative activity in these groups, enzyme-linked immunosorbent assay （ELISA） to measure the level of IL-1β protein in the culture supernatant of HaCaT cells, quantitative fluorescent PCR to detect the mRNA expression of IL-1β in HaCaT cells. Data were statistically analyzed by using one-way analysis of variance （ANOVA） followed by the least significant difference （LSD） test. Results The treatments with ECPIRM of 20 and 40 μmol/L for 24 hours could promote the proliferation of HaCaT cells. However, with the increase in treatment duration and concentrations, ECPIRM inhibited the growth of HaCaT cells in a concentration- and time-dependent manner. ATRA also decelerated the proliferation of HaCaT cells in a concentration- and time-dependent manner. The levels of supernatant IL-1β protein and intracellular IL-1β mRNA were both significantly higher in the IL-17 group than in the control group （IL-1β protein： 20.312 ± 2.053 ng/L vs. 11.427 ± 0.929 ng/L, P 〈 0.05; IL-1β mRNA： 4.05 ± 0.47 vs. 1 ± 0.03, P 〈 0.05）, but significantly lower in the IL-17 ＋ ECPIRM group （12.470 ± 1.897 ng/L and 0.82 ± 0.12 respectively, both P 〈 0.05） and IL-17 ＋ ATRA group （12.694 ± 1.478 ng/L and 0.87 ± 0.16, respectively, both P 〈 0.05�

关 键 词：角蛋白细胞 维生素A酸类 白细胞介素17 白细胞介素1Β HACAT细胞 ECPIRM ATRA 

分 类 号：R751[医药卫生—皮肤病学与性病学]