窄谱中波紫外线通过下调miRNA-25表达促进黑素细胞增殖和黑素生成  

NB-UVB promotes melanogenesis and cell proliferation through downregulating the expression of miRNA-25 in human melanocytes

作　　者：董妍[1] 李文彬 郑雯[1] 王娟[1] 马慧群[1] 曾维惠[1] 

机构地区：[1]西安交通大学第二附属医院,陕西西安710004 [2]陕西省中医医院皮肤科,陕西西安710003

出　　处：《西安交通大学学报（医学版）》2016年第3期399-402,407,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的研究窄谱中波紫外线(NB-UVB)对黑素细胞增殖、黑素生成、酪氨酸酶活性及miRNA-25表达的影响,探讨NB-UVB与miRNA-25间的关系以及NB-UVB治疗白癜风的可能作用机制。方法以40mJ/cm2 NB-UVB作用于体外培养的黑素细胞72h,观察NB-UVB对细胞增殖、酪氨酸酶活性、黑素生成的影响;NB-UVB作用12h后,检测miRNA-25表达的变化。分别转染miRNA-25模拟物、miRNA-25抑制剂和miRNA-25突变体,观察细胞增殖、酪氨酸酶活性、黑素生成的变化。使用MTT法检测黑素细胞活性;左旋多巴底物法测定酪氨酸酶活性;NaOH法检测黑素生成,实时荧光定量聚合酶链反应(qRT-PCR)检测miRNA-25表达量。结果 40mJ/cm2剂量的NB-UVB作用细胞72h后,黑素细胞增殖、酪氨酸酶活性增大,黑素合成显著增加(P<0.05);NB-UVB作用12h后,miRNA-25表达量显著低于对照组(P<0.05)。下调miRNA-25表达后,黑素细胞增殖、酪氨酸酶活性、黑素合成增加;过表达miRNA-25后,黑素细胞增殖、酪氨酸酶活性降低,黑素生成减少;过表达miRNA-25后部分抑制了NB-UVB对黑素细胞的作用。结论 NB-UVB可能部分通过下调黑素细胞miRNA-25的表达,促进黑素细胞增殖、增加酪氨酸酶活性及黑素生成。Objective To investigate the effect of narrow-band UVB （NB-UVB） on melanocytes proliferation, melanin production, tyrosinase activities and miRNA-25 expression so as to explore the relationship between NB-UVB and miRNA-25 and the possible mechanism of NB-UVB for treatment of vitiligo. Methods Melanocytes cultured in vitro were treated with 40 mJ/cm2 dose of NB-UVB for 72 h, the effects of NB-UVB on cell proliferation, tyrosinase activity and melanin content were investigated. After NB-UVB stimulation for 12 h, the effect of NB-UVB on miRNA-25 expression in melanocytes was detected. Melanocytes were transfected with miRNA-25 mimics, miRNA-25 inhibitor and miRNA-25 mutant, respectively, the changes of cell proliferation, tyrosinase activitiy and melanin content were observed. Cell viability was detected using MTT method. Tyrosinase activities were measured with levodopa as the substrate. NaOH assay was used for the detection of melanin content. Real-time quantitative polymerase chain reaction （qRT-PCR） was used to detect miRNA-25 expression. Results After NB-UVB of 40 mJ/cmz dose was used for the cells cultured for 72 h, the viability of melanocytes, tyrosinase activities, and melanin content were significantly increased （P〈0.05）. After NB-UVB stimulation for 12 h, miRNA-25 expression was significantly lower than that in the control group （P^0. 05）. After knockdown of miRNA-25, the cell proliferation of melanocytes, tyrosinase activities and melanin content were increased, whereas overexpression of miRNA-25 decreased the cell proliferation, tyrosinase activities and melanin production.Overexpression of miRNA-25 partially inhibited the effect of NB-UVB on the treatment of melanocytes.Conclusion NB-UVB may promote cell proliferation, increase tyrosinase activities and melanin formation through partially downregulating the expression of miRNA-25 in melanocytes.

关 键 词：窄谱中波紫外线 miRNA-25 黑素细胞 黑素生成 

分 类 号：R758.4[医药卫生—皮肤病学与性病学]