中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (11): 802-805.doi: 10.3760/cma.j.issn.0412-4030.2018.11.005

• 论著 • 上一篇    下一篇

甘草次酸对银屑病患者角质形成细胞增殖、凋亡及miR-21表达的影响

郑瑀心 闵敏 朱子羽 杨晓红 曹毅 郑敏   

  1. 浙江中医药大学(郑瑀心、朱子羽、杨晓红、曹毅);江苏省常州市第一人民医院皮肤科(闵敏);浙江大学医学院附属第二医院皮肤科(郑敏)
  • 收稿日期:2018-02-24 修回日期:2018-08-21 发布日期:2018-10-31
  • 通讯作者: 曹毅 E-mail:caoyi1965@163.com
  • 基金资助:
    2017年度浙江省大学生科技创新活动计划暨新苗人才计划项目(2017R410002)

Effects of glycyrrhetinic acid on proliferation and apoptosis of, as well as miR-21 xpression in keratinocytes from patients with psoriasis

Zheng Yuxin, Min Min, Zhu Ziyu, Yang Xiaohong, Cao Yi, Zheng Min   

  1. Zhejiang Chinese Medical University, Hangzhou 310053, China (Zheng YX, Zhu ZY, Yang XH, Cao Y); Department of Dermatology, The First People′s Hospital of Changzhou, Changzhou 213003, Jiangsu, China (Min M); Department of Dermatology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China (Zheng M)
  • Received:2018-02-24 Revised:2018-08-21 Published:2018-10-31
  • Contact: Cao Yi E-mail:caoyi1965@163.com
  • Supported by:
    Zhejiang Xin?miao Talents Program and National College Students Innovation and Entrepreneurship Program in 2017 (2017R410002)

摘要: 目的 初步探讨甘草次酸对银屑病患者角质形成细胞增殖、凋亡的影响以及可能的作用机制。方法 体外原代培养银屑病患者角质形成细胞,将传代2 ~ 3次后的角质形成细胞分为对照组和药物处理组,药物处理组加入甘草次酸,使其终浓度分别为1、2、4、8和10 mg/L,对照组不加甘草次酸。作用24 ~ 72 h后,MTS法检测甘草次酸对角质形成细胞增殖的影响;流式细胞仪检测不同浓度甘草次酸处理24 h后,角质形成细胞凋亡的变化;实时荧光定量PCR检测甘草次酸对银屑病患者角质形成细胞miR-21表达的影响。结果 1 ~ 10 mg/L甘草次酸作用角质形成细胞24 ~ 72 h后,随着甘草次酸作用时间延长和浓度增加,细胞增殖率逐渐下降。1、2、4、8和10 mg/L甘草次酸作用24 h后,角质形成细胞凋亡率分别上升至(9.64 ± 0.86)%、(25.24 ± 2.93)%、(27.68 ± 3.70)%、(35.55 ± 4.23)%、(38.89 ± 2.31)%;两两比较显示,除1 mg/L甘草次酸组外,其他各组与对照组(10.09 ± 0.69)%相比,差异均有统计学意义(均P < 0.01)。1、2、4 mg/L甘草次酸处理角质形成细胞24 h后,miR-21表达水平(2-ΔΔCt)分别为0.24 ± 0.04、0.22 ± 0.07、0.17 ± 0.05,与对照组0.92 ± 0.12相比,差异有统计学意义(F = 213.10,P < 0.05)。2 mg/L甘草次酸作用18、24、48 h后,miR-21基因表达水平分别为0.55 ± 0.02、0.22 ± 0.06、0.15 ± 0.06,总体呈下降趋势,与对照组(0.98 ± 0.02)相比,差异有统计学意义(F = 238.10,P < 0.05)。结论 甘草次酸可抑制银屑病患者角质形成细胞的增殖,促进其凋亡,其机制可能与下调miR-21表达有关。

关键词: 银屑病, 甘草次酸, 角蛋白细胞, 细胞增殖, 细胞凋亡, 微RNAs, miRNA?21

Abstract: Zheng Yuxin, Min Min, Zhu Ziyu, Yang Xiaohong, Cao Yi, Zheng Min Zhejiang Chinese Medical University, Hangzhou 310053, China (Zheng YX, Zhu ZY, Yang XH, Cao Y); Department of Dermatology, The First People′s Hospital of Changzhou, Changzhou 213003, Jiangsu, China (Min M); Department of Dermatology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China (Zheng M) Corresponding author: Cao Yi, Email: caoyi1965@163.com 【Abstract】 Objective To preliminarily evaluate the effect of glycyrrhetinic acid on the proliferation and apoptosis of keratinocytes in patients with psoriasis, and to explore its possible mechanisms. Methods Keratinocytes were isolated from patients with psoriasis, and subjected to a primary culture in vitro. After 2 - 3 passages, the keratinocytes were divided into several groups to be treated with glycyrrhetinic acid at final concentrations of 0 (control group), 1, 2, 4, 8 and 10 mg/L (glycyrrhetinic acid groups), respectively. After 24 - 72 hours of treatment, MTS assay was performed to evaluate the effect of glycyrrhetinic acid on the proliferation of keratinocytes, and flow cytometry was conducted to detect the apoptosis of keratinocytes after 24-hour treatment with glycyrrhetinic acid at different concentrations. Real-time fluorescence-based PCR was performed to determine the expression of miR-21 in keratinocytes. Results After 24 - 72 hours of treatment with 1 - 10 mg/L glycyrrhetinic acid, the proliferation activity of keratinocytes significantly decreased along with the increase in the treatment duration and concentrations of glycyrrhizinic acid. After 24-hour treatment with 1, 2, 4, 8, 10 mg/L glycyrrhetinic acid, the apoptosis rates of keratinocytes increased to (9.64 ± 0.86)%, (25.24 ± 2.93)%, (27.68 ± 3.70)%, (35.55 ± 4.23)% and (38.89 ± 2.31)% respectively. As LSD-t test showed, the apoptosis rates of keratinocytes were significantly higher in all the glycyrrhetinic acid groups than in the control group (10.09% ± 0.69%, all P < 0.01), except the 1 mg/L glycyrrhetinic acid group. After 24-hour treatment with 1, 2 and 4 mg/L glycyrrhetinic acid, the miR-21 expression (2-ΔΔCt) significantly decreased (0.24 ± 0.04, 0.22 ± 0.07, 0.17 ± 0.05, respectively) compared with the control group (0.92 ± 0.12, F = 213.10, P < 0.05). After 18-, 24- and 48-hour treatment with 2 mg/L glycyrrhetinic acid, the miR-21 expression significantly decreased (0.55 ± 0.02, 0.22 ± 0.06 and 0.15 ± 0.06 respectively) compared with the control group (0.98 ± 0.02, F = 238.10, P < 0.05). Conclusion Glycyrrhetinic acid can inhibit the proliferation of keratinocytes from psoriatic patients, but promote the apoptosis, likely by down-regulation of miR-21 expression.

Key words: Psoriasis, Glycyrrhetinic acid, Keratinocytes, Cell proliferation, Apoptosis, MicroRNAs, miR?21

引用本文

郑瑀心 闵敏 朱子羽 杨晓红 曹毅 郑敏. 甘草次酸对银屑病患者角质形成细胞增殖、凋亡及miR-21表达的影响[J]. 中华皮肤科杂志, 2018,51(11):802-805. doi:10.3760/cma.j.issn.0412-4030.2018.11.005

Zheng Yuxin, Min Min, Zhu Ziyu, Yang Xiaohong, Cao Yi, Zheng Min. Effects of glycyrrhetinic acid on proliferation and apoptosis of, as well as miR-21 xpression in keratinocytes from patients with psoriasis[J]. Chinese Journal of Dermatology, 2018, 51(11): 802-805.doi:10.3760/cma.j.issn.0412-4030.2018.11.005