中华皮肤科杂志 ›› 2020, Vol. 53 ›› Issue (6): 452-458.doi: 10.35541/cjd.20190749

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【已撤稿】丹参素对银屑病样细胞Yes相关蛋白表达及细胞增殖、凋亡的影响

贾金靖1，2    莫秀梅2    刘俊峰2    王宁1    郑焱1    陈达灿2   

  1. 1西安交通大学第二附属医院皮肤科  710004；2广州中医药大学第二附属医院皮肤科  510120
  • 收稿日期:2019-07-16 修回日期:2019-11-22 发布日期:2020-06-01
  • 通讯作者: 陈达灿 E-mail:4910702@163.com
  • 基金资助:
    陕西省自然科学基础研究计划项目（2018JQ8067）；广东省中医药科技创新平台建设项目（粤中医药函［2017］340号）

【Retraction】Effect of danshensu on Yes-associated protein expression in as well as on proliferation and apoptosis of a psoriasis-like cell model

Jia Jinjing1,2, Mo Xiumei2, Liu Junfeng2, Wang Ning1, Zheng Yan1, Chen Dacan2   

  1. 1Department of Dermatology, The Second Affiliated Hospital of Xi′an Jiaotong University, Xi′an 710004, China; 2Department of Dermatology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China
  • Received:2019-07-16 Revised:2019-11-22 Published:2020-06-01
  • Contact: Chen Dacan E-mail:4910702@163.com
  • Supported by:
    Natural Science Basic Research Program of Shaanxi Province-General Project （Youth） （2018JQ8067）; Guangzhou Traditional Chinese Medicine Science and Technology Innovation Platform Construction Project （Guangdong Traditional Chinese Medicine Bureau ［2017］ 340）

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摘要：

此文已撤稿

【摘要】 目的 研究丹参素对M5刺激的HaCaT细胞（银屑病样细胞模型）增殖、凋亡的影响及与Yes相关蛋白（YAP）的关系。方法 采用M5（含10 μg/L的IL-1α、IL-17、IL-22、TNF-α、Oncostatin M）作用于HaCaT细胞48 h，制作银屑病样细胞模型，同时分别加入0（对照组）、0.125、0.25、0.5 mmol/L丹参素刺激细胞，以未做处理的HaCaT细胞为空白对照组。实时定量PCR及Western印迹检测各组细胞YAP的表达；MTT法检测丹参素刺激细胞24、48、72 h时各组细胞的增殖活性；流式细胞仪检测丹参素对各组细胞周期及凋亡的影响，Western印迹检测各组细胞周期相关蛋白（cyclin A、cyclin B1、cyclin D、cyclin E）及凋亡相关蛋白（cleaved caspase-3、Bcl-2、BAX、p53、p21）的表达。多组间均数比较采用单因素方差分析，组间两两比较用LSD-t检验。结果 空白对照组与0（对照组）、0.125、0.25、0.5 mmol/L丹参素组细胞YAP mRNA、蛋白的相对表达水平及培养24、48、72 h时的增殖活力差异均有统计学意义（P < 0.001），0.125、0.25、0.5 mmol/L丹参素组YAP mRNA（1.76 ± 0.04、1.54 ± 0.05、1.33 ± 0.05）、YAP蛋白的相对表达水平（1.78 ± 0.06、1.49 ± 0.32、1.27 ± 0.04）及培养48 h时的细胞增殖活力（1.66 ± 0.04、1.52 ± 0.02、1.34 ± 0.04）均低于对照组（mRNA：2.04 ± 0.04，蛋白：2.10 ± 0.06，活力：1.82 ± 0.03，均P < 0.05）。流式细胞仪检测显示，5组细胞G0/G1期、S期、G2/M期细胞比例及凋亡率差异均有统计学意义（P < 0.001），0.125、0.25、0.5 mmol/L丹参素组细胞G0/G1期、G2/M期细胞比例高于对照组，S期细胞比例低于对照组（均P < 0.05），0.25、0.5 mmol/L丹参素组细胞凋亡率高于对照组（均P < 0.05）。Western印迹显示，5组的细胞周期相关蛋白及凋亡相关蛋白相对表达差异均有统计学意义。结论 丹参素可能通过抑制YAP的表达而抑制银屑病样细胞的增殖与促进其凋亡。

关键词: 银屑病, 丹参素, 角蛋白细胞, 细胞增殖, 细胞凋亡, Yes相关蛋白

Abstract:

【Retraction】

【Abstract】 Objective To evaluate the effect of danshensu on proliferation and apoptosis of M5-stimulated HaCaT cells （a psoriasis-like cell model）, and to explore its relationship with Yes-associated protein （YAP） expression. Methods HaCaT cells were stimulated with M5, a mixture containing 10 μg/L interleukin（IL）-1α, IL-17, IL-22, tumor necrosis factor （TNF）-α and oncostatin M, for 48 hours to establish a psoriasis-like cell model. Then, the cell model was divided into several groups to be treated with 0 （control group）, 0.125, 0.25 and 0.5 mmol/L danshensu respectively, and HaCaT cells receiving no treatment served as the blank control group. Real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of YAP respectively in these groups; methyl thiazolyl tetrazolium （MTT） assay was conducted to estimate the cellular proliferative activity after 24-, 48- and 72-hour treatment with danshensu, flow cytometry to evaluate the effect of danshensu on cell cycle and apoptosis, and Western blot analysis to determine expression of cell cycle-related proteins （cyclin A, cyclin B1, cyclin D, cyclin E） and apoptosis-related proteins （cleaved caspase-3, Bcl-2, BAX, p53 and p21）. One-way analysis of variance was used for comparing means in several groups, and least significant difference （LSD）-t test for multiple comparisons. Results The mRNA and protein expression of YAP significantly differed among the blank control group, control group, 0.125-, 0.25- and 0.5-mmol/L danshensu groups （both P < 0.001）, so did the cellular proliferative activity at 24, 48 and 72 hours （all P < 0.001）. The 0.125-, 0.25- and 0.5-mmol/L danshensu groups all showed significantly decreased mRNA and protein expression of YAP （mRNA: 1.76 ± 0.04, 1.54 ± 0.05, 1.33 ± 0.05 respectively; protein: 1.78 ± 0.06, 1.49 ± 0.32, 1.27 ± 0.04 respectively）, and cellular proliferative activity at 48 hours （1.66 ± 0.04, 1.52 ± 0.02, 1.34 ± 0.04 respectively） compared with the control group （mRNA: 2.04 ± 0.04; protein: 2.10 ± 0.06; cellular proliferative activity: 1.82 ± 0.03; all P < 0.05）. Flow cytometry showed significant differences in the proportions of cells at G0/G1, S and G2/M phases as well as in the apoptosis rates among the above 5 groups （all P < 0.001）. Compared with the control group, the 0.125-, 0.25- and 0.5-mmol/L danshensu groups showed significantly higher proportions of cells at G0/G1 and G2/M phases, but lower proportions of cells at S phase （all P < 0.05）. Additionally, the apoptosis rates were significantly higher in the 0.25- and 0.5-mmol/L danshensu groups than in the control group （both P < 0.05）. Western blot analysis revealed significant differences in the expression of cell cycle-related proteins and apoptosis-related proteins among the above 5 groups （all P < 0.001）. Conclusion Danshensu can inhibit the proliferation of the psoriasis-like cell model and promote its apoptosis, likely by suppressing YAP expression.

Key words: Psoriasis, Danshensu, Keratinocytes, Cell proliferation, Apoptosis, Yes-associated protein

引用本文

贾金靖 莫秀梅 刘俊峰 王宁 郑焱 陈达灿. 【已撤稿】丹参素对银屑病样细胞Yes相关蛋白表达及细胞增殖、凋亡的影响[J]. 中华皮肤科杂志, 2020,53(6):452-458. doi:10.35541/cjd.20190749

Jia Jinjing, Mo Xiumei, Liu Junfeng, Wang Ning, Zheng Yan, Chen Dacan. 【Retraction】Effect of danshensu on Yes-associated protein expression in as well as on proliferation and apoptosis of a psoriasis-like cell model[J]. Chinese Journal of Dermatology, 2020, 53(6): 452-458.doi:10.35541/cjd.20190749