吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (4): 965-970.doi: 10.13481/j.1671-587X.20210420

• 基础研究 • 上一篇    下一篇

薄芝糖肽对人角质细胞增殖和黏附功能的影响及其治疗银屑病的机制

马振卉1,2,吕淑莲3,田亚萍1()   

  1. 1.吉林大学第一医院皮肤科,吉林 长春 130021
    2.山东省临沂市人民医院皮肤科,山东 临沂 276000
    3.中国人民解放军联勤保障部队第九六四医院南湖院区皮肤科,吉林 长春 130021
  • 收稿日期:2021-02-02 出版日期:2021-07-28 发布日期:2021-07-22
  • 通讯作者: 田亚萍 E-mail:typ@jlu.edu.cn
  • 作者简介:马振卉 (1991-),女,山东省滨州市人,住院医师,医学硕士,主要从事皮肤性病学方面的研究。
  • 基金资助:
    吉林省科技厅科技发展计划项目(20190201049JC);吉林省卫健委卫生与健康技术创新项目(2018J056)

Effects of Bozhi glycopeptide on proliferation and adhesion function of human keratinocytes and its mechanism in treatment of psoriasis

Zhenhui MA1,2,shulian LYU3,Yaping TIAN1()   

  1. 1.Department of Dermatology and Venerology,First Hospital,Jilin University,Changchun 130021,China
    2.Department of Dermatology and Venerology,Linyi People’s Hospital,Shandong Province,Linyi 276000,China
    3.Department of Dermatology and Venerology,No. 964 Hospital,PLA Joint Logistics Support Force,Changchun 130021,China
  • Received:2021-02-02 Online:2021-07-28 Published:2021-07-22
  • Contact: Yaping TIAN E-mail:typ@jlu.edu.cn

摘要: 目的

探讨薄芝糖肽对人角质细胞(KC)增殖和黏附因子β-连环蛋白(β-catenin)表达的影响,阐明薄芝糖肽治疗银屑病的可能机制。

方法

增殖功能实验中,对数生长期的人永生化角质形成细胞(HaCaT细胞)分为对照组和不同浓度(0.011 719、0.023 438、0.046 875、0.093 750、0.187 500、0.375 000、0.750 000和1.500 000 g·L-1)薄芝糖肽组,分别处理24 h后,采用CCK-8法检测各组细胞增殖率,确定薄芝糖肽作用的有效浓度;对数生长期HaCaT细胞分为对照组和不同浓度(0.625、1.250、2.500、5.000、10.000、20.000、40.000和80.000 mg·L-1)薄芝糖肽组,分别处理24和48 h后,采用CCK-8法检测各组细胞增殖率。黏附功能实验中,对数生长期HaCaT细胞分为对照组和不同浓度(10和20 mg·L-1)薄芝糖肽组,分别处理24和48 h后,实时荧光定量PCR(RT-qPCR)法检测各组细胞中β-catenin mRNA表达水平,Western blotting法检测各组细胞中β-catenin蛋白表达水平。

结果

增殖功能实验,与对照组比较,作用24 h后,0.011 719~0.093 750 g·L-1薄芝糖肽组细胞增殖率均明显降低(P<0.01),该浓度区间为薄芝糖肽有效浓度区间。与对照组比较,作用24和48 h后, 1.250、2.500、5.000、10.000、20.000、40.000和80.000 mg·L-1薄芝糖肽组细胞增殖率均明显降低(P<0.05或P<0.01),且20 mg·L-1薄芝糖肽组作用48 h时细胞增殖率降低最明显(P<0.01)。黏附功能实验,与对照组比较,作用24 h时10和20 mg·L-1薄芝糖肽组细胞中β-catenin mRNA表达水平明显降低(P<0.01),作用48 h时10和20 mg?L-1薄芝糖肽组细胞中β-catenin mRNA表达水平降低更明显(P<0.01);与对照组比较,作用24 h时10 mg?L-1薄芝糖肽组细胞中β-catenin蛋白表达水平明显降低(P<0.01),作用48 h时10和20 mg?L-1薄芝糖肽组细胞中β-catenin蛋白表达水平有增高趋势,但差异无统计学意义(P>0.05)。

结论

薄芝糖肽在一定的浓度和作用时间内可以抑制KC增殖,并可通过降低β-catenin表达抑制KC黏附功能,可能是薄芝糖肽治疗银屑病的作用机制之一。

关键词: 薄芝糖肽, 银屑病, 人角质细胞, β-连环蛋白, 细胞增殖, 细胞黏附

Abstract: Objective

To investigate the effects of Bozhi glycopeptide on the proliferation of the human keratinocytes (KC) and the expression of β-catenin, and to elucidate the possible mechanism of Bozhi glycopeptide in the treatment of psoriasis.

Methods

In the proliferation function experiment, the HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (0.011 719, 0.023 438, 0.046 875, 0.093 750, 0.187 500, 0.375 000, 0.750 000 and 1.500 000 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24h, the CCK-8 method was used to detect the proliferation rates of the cells in various groups to determine the effective concentration of Bozhi glycopeptide. The HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (0.625, 1.250, 2.500, 5.000, 10.000, 20.000, 40.0000 and 80.000 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24 and 48 h, the cell proliferation rates in various groups were detected by CCK-8 method. In the adhesion function experiment, the HaCaT cells in the logarithmic growth phase were divided into control group and different concentrations (10 and 20 mg·L-1) of Bozhi glycopeptide groups. After the cells were treated for 24 and 48 h, the RT-qPCR method was used to detect the expression levels of β-catenin mRNA in the cells in various groups; Western blotting method was used to detect the expression levels of β-catenin protein in the cells in various groups.

Results

In the proliferation function experiment, compared with control group, the cell proliferation rates in 0.011 719-0.093 750 g·L-1 Bozhi glycopeptide groups were significantly reduced after treated for 24 h(P<0.05), which was the effective concentration range of Bozhi glycopeptide. Compared with control group,the proliferation rates of cells in 1.250, 2.500,5.000,10.000,20.000,40.000,and 80.000 mg·L-1 Bozhi glycopeptide groups were significantly reduced after treated for 24 and 48 h(P<0.05 or P<0.01); the proliferation rate was decreased significantly (P<0.01) when the cells were treated with 20 mg·L-1 Bozhi glycopeptide for 48 h. In the adhesion function experiment, compared with control group, the expression levels of β-catenin mRNA in the cells in 10 and 20 mg·L-1 Bozhi glycopeptide groups were decreased significantly after treated for 24 h (P<0.01), and the expression levels of β-catenin mRNA were decreased significantly after treated for 48 h (P<0.01). Compared with control group, after 24 h of treatment, the expression level of β-catenin protein in 10 mg·L-1 Bozhi glycopeptide group was decreased significantly; after 48 h of treatment, the expression levels of β-catenin protein in 10 and 20 mg·L-1 Bozhi glycopeptide groups tended to increase,but there were no significant differences(P>0.05).

Conclusion

Bozhi glycopeptide can inhibit the proliferation of KC at a certain concentration and action time, and can inhibit KC adhesion function by reducing the expression of β-catenin. This may be one of the possible mechanisms of Bozhi glycopeptide in the treatment of psoriasis.

Key words: Bozhi glycopeptide, psoriasis, human keratinocytes, β-catenin, cell proliferation, cell adhesion

中图分类号: 

  • R285.5