异欧前胡素对体外人表皮黑素细胞酪氨酸酶和β-肌动蛋白表达的调节作用

doi:10.3969/j.issn.1004-616x.2017.05.008

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (5): 364-369.doi: 10.3969/j.issn.1004-616x.2017.05.008

异欧前胡素对体外人表皮黑素细胞酪氨酸酶和β-肌动蛋白表达的调节作用

詹标¹, 孙贝蕾², 罗思聪¹, 庄鸿凯¹, 黄鹏洲¹, 曾懿¹, 陈玮芳³, 陈玮莹²

1. 汕头大学医学院2013级7年制临床医学专业, 广东汕头 515041;
2. 汕头大学医学院基础医学实验教学中心, 广东汕头 515041;
3. 汕头大学医学院第二附属医院, 广东汕头 515041

收稿日期:2016-09-25 修回日期:2017-01-06 出版日期:2017-09-30 发布日期:2017-09-30
通讯作者: 陈玮芳,E-mail:cwflxk2@139.com;陈玮莹,E-mail:wychen@stu.edu.cn E-mail:cwflxk2@139.com;wychen@stu.edu.cn
作者简介:詹标,E-mail:13bzhan@stu.edu.cn;孙贝蕾,E-mail:923719157@qq.com。
基金资助:
教育部大学生创新训练项目（201510560032）；广东省中医药局项目（20142082）

Regulatory effects of isoimperatorin on expression of tyrosinase and β-actin in cultured human epidermal melanocytes

ZHAN Biao¹, SUN Beilei², LUO Sicong¹, ZHUANG Hongkai¹, HUANG Pengzhou¹, ZENG Yi¹, CHEN Weifang³, CHEN Weiying²

1. 2013 Students of Seven-year Clinical Medicine Program, Shantou University Medical College, Shantou 515041;
2. Experimental Teaching Center for Preclinical Medicine, Shantou University Medical College, Shantou 515041;
3. The Second Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2016-09-25 Revised:2017-01-06 Online:2017-09-30 Published:2017-09-30

摘要/Abstract

摘要： 目的：研究异欧前胡素对体外培养的人表皮黑素细胞酪氨酸酶和β-actin蛋白表达的影响。方法：体外分离、纯化培养人表皮黑素细胞，随机分为对照组和异欧前胡素实验组。对照组用M-254培养基培养，实验组加入含25 μmol/L异欧前胡素的M-254 培养基，培养48 h。倒置显微镜下观察人表皮黑素细胞形态，NaOH裂解法检测黑素含量的变化，采用实时荧光定量PCR方法、Western blot方法、免疫荧光法检测酪氨酸酶和β-actin在黑素细胞中的表达及其变化。结果：异欧前胡素作用48 h后，与对照组黑素细胞比较，实验组黑素细胞的树状突起变长，黑素含量减少（P < 0.01）；实时荧光定量PCR、Western blot和免疫荧光法检测结果表明，与对照组相比，实验组黑素细胞酪氨酸酶mRNA及其蛋白的表达降低（P均 < 0.05），β-actin mRNA及其蛋白的表达增加（P均 < 0.05）。结论：25 μmol/L异欧前胡素抑制体外培养人表皮黑素细胞酪氨酸酶mRNA及其蛋白的表达，促进β-actin mRNA及其蛋白的表达，从而降低体外人表皮黑素细胞黑素的含量。

关键词: 黑素细胞, 异欧前胡素, 酪氨酸酶, β-肌动蛋白

Abstract: OBJECTIVE: To investigate the effects of isoimperatorin on expression of tyrosinase and β-actin in cultured human epidermal melanocytes. METHODS: Human epidermal melanocytes were separated,purified and cultured in vitro. These cultures were randomly divided into control and isoimperatorin groups. Cells were cultured with M-254 medium. For the isoimperatorin group,25 μmol/L isoimperatorin in M-254 was added to the cultures. After 48 h of incubation,the morphology of melanocytes was observed under an inverted microscope. Melanin content was assayed by using the NaOH method. The methods of qPCR,Western blot and immunofluorescent were used to analyze the expression of tyrosinase and β-actin. RESULTS: After treatment with 25 μmol/L isoimperatorin for 48 h,dendrites of melanocytes were slightly elongated compared with the untreated cells. In addition,the treatment reduced melanin content (P < 0.01). The results of qPCR,Western blot and immunofluorescence assay consistently indicated that isoimperatorin reduced tyrosinase mRNA and protein,and increased β-actin mRNA and protein. CONCLUSION: Cultured human epidermal melanocytes which were treated with 25 μmol/L isoimperatorin had reduced content of melanin. The observed effect was associated with inhibition of tyrosinase mRNA and protein,and with increased β-actin mRNA and protein.

Key words: melanocyte, isoimperatorin, tyrosinase, β-actin

中图分类号:

R329.2

詹标, 孙贝蕾, 罗思聪, 庄鸿凯, 黄鹏洲, 曾懿, 陈玮芳, 陈玮莹. 异欧前胡素对体外人表皮黑素细胞酪氨酸酶和β-肌动蛋白表达的调节作用[J]. 癌变·畸变·突变, 2017, 29(5): 364-369.

ZHAN Biao, SUN Beilei, LUO Sicong, ZHUANG Hongkai, HUANG Pengzhou, ZENG Yi, CHEN Weifang, CHEN Weiying. Regulatory effects of isoimperatorin on expression of tyrosinase and β-actin in cultured human epidermal melanocytes[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 364-369.

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异欧前胡素对体外人表皮黑素细胞酪氨酸酶和β-肌动蛋白表达的调节作用

Regulatory effects of isoimperatorin on expression of tyrosinase and β-actin in cultured human epidermal melanocytes

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