微小RNA-27a-3p对人表皮角质形成细胞损伤的影响  

Effects of microRNA-27a-3p on the damage of human epidermal keratinocytes

作　　者：郑树茂 王华 白帆 田赞 张莉 ZHENG Shu-mao;WANG Hua;BAI Fan;TIAN Zan;ZHANG Li(Department of Dermatology,Hebei Academy of Traditional Chinese Medicine,Shijiazhuang 050000,Hebei Province,China;First Department of Dermatology,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,Hebei Province,China;Department of Dermatology,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,Hebei Province,China;Department of Spleen and Stomach,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,Hebei Province,China)

机构地区：[1]河北省中医药科学院皮肤科,河北石家庄050000 [2]河北省中医院皮肤一科,河北石家庄050011 [3]河北省中医院皮肤科,河北石家庄050011 [4]河北省中医院脾胃科,河北石家庄050011

出　　处：《中国临床药理学杂志》2021年第19期2676-2679,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究微小RNA-27a-3p(miR-27a-3p)对白细胞介素-22(IL-22)诱导的人表皮角质形成细胞损伤的影响。方法取人表皮角质形成细胞NHEK细胞并随机分为对照组,模型组,miR-NC组和miR-27a-3p mimic组。对照组为正常条件培养的NHEK细胞;模型组用100 ng·mL^(-1)IL-22处理NHEK细胞24 h;miR-NC组和miR-27a-3p mimic组分别转染阴性对照寡核苷酸和miR-27a-3p模拟物,随后用IL-22处理NHEK细胞24 h。用细胞计数试剂盒-8(CCK-8)检测各组光密度并计算细胞存活率,以流式细胞术检测各组细胞凋亡率,以双荧光素酶报告实验验证miR-27a-3p与YAP1靶向关系,以蛋白质印迹法检测白细胞介素-8(Interleukin-8,IL-8),白细胞介素-1β(IL-1β)与Yes相关蛋白1 (YAP1)蛋白表达量。结果对照组、模型组、miR-NC组和miR-27a-3p mimic组48 h时细胞存活率分别为(100.00±0.00)%,(65.96±0.87)%,(67.32±1.26)%,(74.95±0.94)%;72h时细胞存活率分别(100.00±0.00)%,(62.95±0.75)%,(64.30±1.06)%,(69.90±0.89)%;细胞凋亡率分别为(7.03±0.74)%,(14.13±1.55)%,(13.16±1.65)%,(10.06±1.76)%;以上所有数据,模型组与对照组相比,miR-27a-3p mimic组与miR-NC组相比,差异均有统计学意义(均P <0.05)。此外,双荧光素酶报告实验表明miR-27a-5p可靶向YAP1。结论 miR-27a-3p可抑制IL-22诱导人表皮角质形成细胞NHEK细胞中炎症因子的表达和细胞凋亡,其机制可能与抑制YAP1蛋白表达有关。Objective To investigate the effects of miR-27a-3p on the injury of human epidermal keratinocytes(NHEKs) induced by interleukin-22(IL-22).Methods NHEK cells were randomly divided into control group,model group,miR-NC group and miR-27a-3p mimic group.Control group was cultured under normal conditions;model group was treated with 100 ng·mL^(-1) IL-22 for 24 h;miR-NC group and miR-27a-3p mimic group were transfected with negative control oligonucleotides and miR-27a-3p mimics,respectively,followed by IL-22 treatment for 24 h.Cell counting kit-8(CCK-8)assay was performed to detect the optical dersity of each group and calculate cell survival rate.The apoptosis rate of each group was detected by flow cytometry.The relationship between miR-27a-3p and YAP1 was verified by dual-luciferase reporter assay.Western blot was performed to detect the protein expression levels of interleukin-8(IL-8),interleukin-1β(IL-1β) and Yes-associated protein 1(YAP1).Results The cell survival rates at 48 h of control,model,miR-NC,miR-27a-3p mimic groups were(100.00±0.00) %,(65.96±0.87) %,(67.32±1.26) % and(74.95±0.94) %;the cell survival rates at 72 h were(100.00±0.00) %,(62.95±0.75) %,(64.30±1.06) % and(69.90±0.89) %;the cell apoptosis rates were(7.03±0.74) %,(14.13±1.55) %,(13.16±1.65) % and(10.06±1.76) %.In terms of above indexes,the differences were statistically significant between model group and control group,as well as between miR-27a-3p mimic group and miR-NC group(P <0.05).In addition,dual-luciferase reporter assay showed that miR-27 a-5 p could target YAP1.Conclusion MiR-27a-3p can inhibit the expressions of inflammatory factors in NHEK cells induced by IL-22 and the cell apoptosis,and its mechanism may be related to the inhibition of YAP1 protein expression.

关 键 词：微小RNA-27a-3p Yes相关蛋白1 角质形成细胞 炎症 凋亡 

分 类 号：R97[医药卫生—药品]