microRNA-125a在寻常型银屑病皮损中的表达及其对HaCaT细胞增殖的影响  

Expression of microRNA-125a in psoriasis vulgaris lesions and its effect on the proliferation of HaCaT cells

作　　者：苏芳[1] 晋亮 刘玮 蔡宏 孙晓冬[1] 孙晓杰[1] 徐桂娟 王强[1] Su Fang;Jin Liang;Liu Wei;Cai Hong;Sun Xiaodong;Sun Xiaojie;Xu Guijuan;Wang Qiang(Department of Dermatology,The Seventh People′s Hospital of Shenyang,Shenyang 110001,China;Department of Dermatology,Air Force Medical Center,Beijing 100142,China)

机构地区：[1]沈阳市第七人民医院皮肤科,110003 [2]解放军空军特色医学中心皮肤科,北京100142

出　　处：《中华皮肤科杂志》2020年第11期886-890,共5页Chinese Journal of Dermatology

基　　金：辽宁省科学技术计划项目(2019-ZD-0975)。

摘　　要：目的检测寻常型银屑病皮损中microRNA-125a(miR-125a)的表达与皮损处炎症因子水平的相关性及其对人永生化角质形成细胞(HaCaT)增殖的影响。方法收集2017—2018年沈阳市第七人民医院40例寻常型银屑病患者皮损及相邻非皮损组织,采用反转录实时荧光定量PCR检测组织中miR-125a的表达及皮损组织中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6、IL-17 mRNA的表达。将miR-125a过表达质粒、过表达对照质粒、miR-125a干扰质粒、干扰对照质粒转染HaCaT细胞,在转染后0、24、48、72 h采用CCK8法检测各组HaCaT细胞增殖能力,采用双抗体夹心酶联免疫吸附法(ELISA)检测质粒转染后HaCaT细胞上清液中TNF-α、IL-1β、IL-6、IL-17水平。相关性分析采用Spearman等级相关检验分析,两组间均数比较采用t检验。结果寻常型银屑病皮损区miR-125a相对表达水平(2-ΔΔCt,0.389±0.354)低于非皮损区(1.106±0.396,t=7.717,P<0.001)。银屑病皮损组织中miR-125a表达与TNF-α、IL-1β、IL-17 mRNA的表达呈负相关(r=-0.447、-0.424、-0.436,均P<0.01)。转染相应质粒后0、24 h时,细胞增殖能力在过表达miR-125a组与过表达对照组(t=0.282、1.445,均P>0.05)、干扰miR-125a组与干扰对照组(t=0.120、1.543,均P>0.05)间差异无统计学意义;转染后48、72 h时,过表达miR-125a组细胞的增殖能力低于过表达对照组(t=3.222、4.563,均P<0.05),干扰miR-125a组高于干扰对照组(t=3.036、3.269,均P<0.05)。MiR-125a过表达组TNF-α、IL-1β的表达水平低于过表达对照组,差异有统计学意义(t=4.318、3.813,均P<0.05)。结论寻常型银屑病患者皮损中miR-125a低表达,miR-125a抑制角质形成细胞增殖,可能在银屑病发生发展中发挥保护作用。Objective To investigate the correlation between microRNA-125a(miR-125a)expression and inflammatory cytokine levels in skin lesions of patients with psoriasis vulgaris,and to evaluate the effect of miR-125a on the proliferation of a human immortalized keratinocyte cell line HaCaT.Methods Totally,lesional and adjacent non-lesional skin tissues were collected from 40 patients with psoriasis vulgaris in the Seventh People′s Hospital of Shenyang from 2017 to 2018,and real-time fluorescence-based quantitative reverse transcription PCR was performed to determine the expression of miR-125a in the skin tissues,as well as the mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and IL-17 in the lesional skin tissues.HaCaT cells were divided into 4 groups to be transfected with a miR-125a overexpression plasmid(miR-125a overexpression group),an overexpression control plasmid(overexpression control group),a miR-125a interference plasmid(miR-125a interference group)and an interference control plasmid(interference control group),respectively.Cell counting kit-8(CCK8)assay was performed to assess the proliferative ability of HaCaT cells in the groups at 0,24,48,72 hours after transfection,and double-antibody sandwich enzyme-linked immunosorbent assay(ELISA)to detect the levels of TNF-α,IL-1β,IL-6 and IL-17 in the culture supernatant of HaCaT cells.Spearman rank correlation test was used for correlation analysis,and t test for the comparison of means between two groups.Results The relative expression of miR-125a was significantly lower in the lesional skin tissues(expressed as 2-ΔΔCt,0.389±0.354)than in the non-lesional skin tissues(1.106±0.396,t=7.717,P<0.001)in patients with psoriasis vulgaris.The expression of miR-125a was negatively correlated with the mRNA expression of TNF-α,IL-1βand IL-17 in psoriatic lesions(r=-0.447,-0.424,-0.436,all P<0.01).Immediately and 24 hours after transfection with the plasmids,there was no significant difference in the cell proliferative ability between th

关 键 词：银屑病 微RNAs 角蛋白细胞 细胞增殖 肿瘤坏死因子α 白细胞介素1β 白细胞介素6 白细胞介素17 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]