中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (2): 104-107.

• 论著 • 上一篇    下一篇

沉默miRNA-23a对UVB诱导成纤维细胞慢性光损伤的影响

张家安1,周炳荣2,胡燕燕3,4,吴维5,尹慧斌6,张倩7,8,郭娴菲2,骆丹9   

  1. 1. 南京医科大学第一附属医院皮肤性病科
    2. 南京医科大学第一附属医院
    3.
    4. 南京医科大学附属第一医院
    5. 南京医科大学附属第一人民医院
    6. 江苏省人民医院
    7. 南京市鼓楼医院病理科
    8. 南京医科大学第一临床医学院
    9. 南京市南京医科大学附属第一医院皮肤科
  • 收稿日期:2013-03-04 修回日期:2013-06-17 出版日期:2014-02-15 发布日期:2014-02-01
  • 通讯作者: 骆丹 E-mail:daniluo2005@163.com
  • 基金资助:
    江苏省自然科学基金;国家自然科学基金;国家自然科学基金

Effect of miRNA-23a silencing on ultraviolet B-induced chronic photodamage to fibroblasts

  • Received:2013-03-04 Revised:2013-06-17 Online:2014-02-15 Published:2014-02-01
  • Supported by:
    Natural Science Foundation of Jiangsu Province of China

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摘要： 【摘要】 目的 探讨沉默miRNA-23a对UVB照射所致成纤维细胞慢性光损伤的影响。 方法 将成纤维细胞分为空白对照组、UVB光损伤组、miRNA-23a 沉默组、UVB光损伤 + miRNA-23a沉默组。SA-β-半乳糖苷酶（SA-β-Gal）化学染色测定老化程度，流式细胞仪检测细胞周期，实时PCR法检测p53、p16、p21 mRNA的表达，免疫印迹法检测p53、p16、p21的蛋白表达量。 结果 UVB光损伤组与UVB光损伤 + miRNA-23a沉默组的SA-β-Gal细胞蓝染阳性率分别为（94.60 ± 2.58）%、（48.18 ± 3.70）%，两组间差异有统计学意义（P < 0.05）。G1期阻滞率分别为（85.06 ± 1.52）%、（57.48 ± 2.01）%，两组间差异有统计学意义（P < 0.05）。UVB光损伤 + miRNA-23a沉默组的p53、p16、p21 mRNA及蛋白表达量与UVB光损伤组相比，差异均有统计学意义（P < 0.05）。 结论 沉默miRNA-23a对UVB诱导成纤维细胞衰老有抑制作用，而miRNA-23a对下游衰老相关信号分子的抑制可能是其机制之一。

关键词: 紫外线, 成纤维细胞, 微RNAs, 细胞衰老

Abstract: Zhang Jiaan, Zhou Bingrong, Hu Yanyan, Wu Wei, Yin Huibin, Zhang Qian, Guo Xianfei, Luo Dan. Department of Dermatology and Venereology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China Corresponding author: Luo Dan, Email: daniluo2011@gmail.com 【Abstract】 Objective To evaluate the effect of miRNA-23a silencing on ultraviolet B （UVB）-induced chronic photodamage to fibroblasts. Methods Fibroblasts from human foreskin were divided into four groups: blank control group receiving untreated, UVB group receiving UVB irradiation only, miRNA-23a group transfected with a miRNA-23a antagomir, UVB + miRNA-23a group receiving transfection with miRNA-23a antagomir followed by UVB irradiation. UVB irradiation was carried out once a day for five consecutive days at a dose of 10 mJ/cm2. Three days after the last irradiation, SA-β-galactosidase staining was performed to detect senescent cells, flow cytometry to analyze cell cycle, and real-time PCR and Western blot to measure the mRNA and protein expressions of p53, p16 and p21, respectively. Results Both the percentage of β-galactosidase-positive cells and proportion of G1-phase cells were significantly higher in the UVB group than in the UVB + miRNA-23a group （（94.60 ± 2.58）% vs. （48.18 ± 3.70）%, （85.06 ± 1.52）% vs. （57.48 ± 2.01）%, both P < 0.05）. Significant differences were observed in the mRNA and protein expressions of p53, p16 and p21 between the UVB group and UVB + miRNA-23a group （all P < 0.05）. Conclusions The silencing of miRNA-23a may suppress UVB-induced chronic photodamage, which is likely to be associated with the inhibition of senescence-associated downstream signaling molecules.

Key words: Ultraviolet rays, Fibroblasts, MicroRNAs, Cell aging