甘草苷对中波紫外线诱导人皮肤角质形成细胞凋亡的影响

张丽宏; 傅云; 廖建; 张琼; 周立禄; 周丽; 王业秋; 李建民

doi:10.13422/j.cnki.syfjx.20180696

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甘草苷对中波紫外线诱导人皮肤角质形成细胞凋亡的影响

药理 | 更新时间：2024-01-04

- 甘草苷对中波紫外线诱导人皮肤角质形成细胞凋亡的影响
- Effect of Liquiritin on UVB-induced Apoptosis of Human Keratinocytes
- 中国实验方剂学杂志 2018年24卷第6期页码：96-101
- 作者机构：
- 作者简介：
- 基金信息：
  
  黑龙江省应用技术研究与开发计划项目（PC13S15）
- DOI：10.13422/j.cnki.syfjx.20180696
  中图分类号： R285
- 纸质出版日期：2018
- 稿件说明：
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张丽宏, 傅云, 廖建, 等. 甘草苷对中波紫外线诱导人皮肤角质形成细胞凋亡的影响[J]. 中国实验方剂学杂志, 2018,24(6):96-101.

ZHANG Li-hong, FU Yun, LIAO Jian, et al. Effect of Liquiritin on UVB-induced Apoptosis of Human Keratinocytes[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(6): 96-101.
张丽宏, 傅云, 廖建, 等. 甘草苷对中波紫外线诱导人皮肤角质形成细胞凋亡的影响[J]. 中国实验方剂学杂志, 2018,24(6):96-101. DOI： 10.13422/j.cnki.syfjx.20180696.

ZHANG Li-hong, FU Yun, LIAO Jian, et al. Effect of Liquiritin on UVB-induced Apoptosis of Human Keratinocytes[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(6): 96-101. DOI： 10.13422/j.cnki.syfjx.20180696.

摘要

目的：研究甘草苷对中波紫外线（UVB）诱导人皮肤角质形成细胞（HaCaT）凋亡的影响。方法：以不同浓度的甘草苷作用于HaCaT细胞，噻唑蓝（MTT）比色法筛选甘草苷的有效安全浓度；用30 mJ · cm-2的UVB作用于HaCaT细胞，建立光老化模型，MTT比色法检测光老化HaCaT细胞增殖率；流式细胞术检测各组细胞中活性氧自由基（ROS）含量和总凋亡率；实时荧光定量PCR （Real-time PCR）检测光老化HaCaT细胞中半胱氨酸天冬氨酸-3（Caspase-3），半胱氨酸天冬氨酸-9（Caspase-9） mRNA表达水平；蛋白免疫印记法（Western blot）检测光老化HaCaT细胞中B淋巴细胞瘤-2（Bcl-2），Caspase-3，Caspase-9蛋白表达量。结果：筛选出甘草苷的最佳有效安全浓度为1×10-7，1×10-6，1×10-5mol · L-1。与空白组比较，UVB组细胞增殖率显著降低（P<0.01），细胞中ROS含量和总凋亡率显著升高（P<0.01），Caspase-3，Caspase-9 mRNA表达水平显著升高（P<0.01）；Bcl-2蛋白表达量显著降低（P<0.01），Caspase-3，Caspase-9蛋白表达量显著升高（P<0.01）。与UVB组比较，1×10-7，1×10-6，1×10-5mol · L-1甘草苷+UVB组细胞增殖率显著升高（P<0.01）；细胞中ROS含量和总凋亡率显著降低（P<0.05，P<0.01），Caspase-3，Caspase-9 mRNA表达水平显著降低（P<0.05，P<0.01）；Bcl-2蛋白表达量显著升高（P<0.05，P<0.01），Caspase-3，Caspase-9蛋白表达量显著降低（P<0.05，P<0.01）。结论：甘草苷能通过促进抗凋亡因子Bcl-2蛋白的表达，减少促凋亡相关细胞因子Caspase-3，Caspase-9 mRNA和蛋白的表达，从而抑制UVB诱导的HaCaT细胞凋亡。

Abstract

Objective: To investigate the effects of liquiritin on ultraviolet B (UVB)-induced apoptosis of human keratinocytes (HaCaT cells).Method: Different concentrations of liquiritinwere used to treat HaCaT cells

and the methyl thiazolyltetrazolium(MTT) assay was used to screen the effective and safe concentration of liquiritin;light aging model was set up by the irradiation of ultraviolet radiation B (UVB) at irradiation dose of 30 mJ · cm-2. Then the activity of cells was determined by MTT method;the content of reactive oxygen species (ROS) and the apoptosis of HaCaT cells were respectively measured by flow cytometry;the expression levels of cysteinyl aspartate-3(Caspase-3) and cysteinyl aspartate-9(Caspase-9) mRNA were determined by Real-time PCR method; and Western blot method was used to detect the effects of liquiritin on B-cell lymphoma 2(Bcl-2)

Caspase-3 and Caspase-9 protein expression.Result: The optimal safe and effective concentrations of liquiritin were 1×10-7

1×10-6

and 1×10-5 mol · L-1. As compared with the blank group

the cell proliferation rate was significantly decreased in the UVB group (P<0.01);the content of ROS and the total apoptosis rate of cells were significantly increased(P<0.01); the expression levels of Caspase-3 and Caspase-9 mRNA were significantly increased (P<0.01);the expression level of Bcl-2 protein was significantly decreased (P<0.01)

and the expression levels of Caspase-3 and Caspase-9 protein were significantly increased (P<0.01). As compared with the UVB group

the cell proliferation rate of the (1×10-7

1×10-6

and 1×10-5) mol · L-1liquiritin+UVB groups were significantly increased (P<0.01);the content of ROS and the total apoptosis rate of cells were significantly decreased (P<0.05

P<0.01); the expression levels of Caspase-3 and Caspase-9 mRNA were significantly decreased(P<0.05

P<0.01);the expression level of Bcl-2 protein was significantly increased(P<0.05

P<0.01)

and the expression levels of Caspase-3 and Caspase-9 protein were significantly decreased (P<0.05

P<0.01).Conclusion: Liquiritin can promote the expression of anti-apoptotic factor Bcl-2 protein

and decrease the expression of the apoptosis-related factors Caspase-3 and Caspase-9 mRNA and protein

thus inhibiting the apoptosis of HaCaT cells induced by UVB.

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