沉默miRNA-23a对UVB诱导成纤维细胞慢性光损伤的影响  

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

作　　者：张家安[1] 周炳荣[1] 胡燕燕[1] 吴维[1] 尹慧彬[1] 张倩[1] 郭娴菲[1] 骆丹[1] 

机构地区：[1]南京医科大学第一附属医院皮肤性病科,210029

出　　处：《中华皮肤科杂志》2014年第2期104-107,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81000700、81171518）;江苏省自然科学基金（BK2012877）

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。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.

关 键 词：紫外线 成纤维细胞 微RNAS 细胞衰老 

分 类 号：R751[医药卫生—皮肤病学与性病学]