miR-142-3p/Sema3A轴调节角质形成细胞增殖凋亡及银屑病皮肤炎症反应  

miR-142-3p/Sema3A Axis Modulates Proliferation and Apoptosis of Keratinocytes and Skin Inflammation in Psoriasis

作　　者：张鼎伟[1] 王媛[1] 霍佳[1] 汪炜[1] 彭斌 张燕飞[1] ZHANG Dingwei;WANG Yuan;HUO Jia;WANG Wei;PENG Bin;ZHANG Yanfei(Department of Dermatology,the Second Affiliated Hospital of Xi′an Jiaotong University Medical College,Xi'an 710004,China)

机构地区：[1]西安交通大学医学院第二附属医院皮肤科,陕西西安710004

出　　处：《中国皮肤性病学杂志》2021年第2期127-132,共6页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金(81703129,81903214)。

摘　　要：目的旨在探讨miR-142-3p在M5刺激人表皮角质形成细胞HaCaT中的作用及潜在机制。方法MTT法检测细胞增殖;ELISA凋亡检测试剂盒评估细胞凋亡。ELISA试剂盒测定炎症介质TNF-α、IL-22和IL-17A的分泌量。RT-PCR和Western blot分别测定目的基因mRNA和蛋白表达。荧光素酶报告实验评估荧光素酶活性。结果M5刺激的HaCaT细胞中miR-142-3p表达升高。下调miR-142-3p显著抑制M5诱导HaCaT细胞的增殖并促进凋亡,抗凋亡蛋白Bcl-2减少,伴随促凋亡蛋白Bax增加。此外,下调miR-142-3p通过降低多种炎症因子的表达改善M5诱导的炎症反应。重要的是,miR-142-3p负调控靶基因Sema3A的表达。从机制上讲,沉默Sema3A有效逆转miR-142-3p下调对HaCaT细胞的抗增殖、促凋亡及抗炎作用。结论下调miR-142-3p通过调控靶基因Sema3A保护HaCaT细胞免受M5诱导的过度增殖和炎症损伤,揭示miR-142-3p/Sema3A轴可能是防止角质形成细胞损伤的新靶点。Objective The aim of the present study was to explore the functions and precise mechanism of miR-142-3 p in human keratinocyte HaCaT cells in the presence of M5.Methods CCK-8 assay was conducted to measure cell proliferation.Apoptosis was assessed using Cell Death Detection ELISAPLUS Kit.The concentrations of TNF-α,IL-22 and IL-17 A were estimated using ELISA assays.RT-PCR and Western blot was applied to evaluate mRNA and protein expression,respectively.Luciferase activity was determined using a Dual-luciferase Reporter Assay.Results MiR-142-3 p expression was heightened in HaCaT cells induced by M5.In addition,inhibition of miR-142-3 p dramatically restricted cell proliferation and enhanced apoptosis in HaCaT cells exposed to M5,as exemplified by a decrease in the anti-apoptotic Bcl-2 protein,concomitant with an increase in the pro-apoptotic proteins Bax.Moreover,depleting miR-142-3 p effectively ameliorated M5-induced inflammation response,as reflected by the attenuation of multiple inflammatory factors.Importantly,Sema3 A was identified as a direct target of miR-142-3 p,and negatively regulated by miR-142-3 p.Mecha-nistically,silencing of Sema3 A effectively abolished the anti-proliferative,apoptosis-promoting,and anti-inflammatory effects of miR-142-3 p inhibition in keratinocytes.Conclusion Repression of miR-142-3 p protected HaCaT cells against M5-induced hyper-proliferation and inflammatory injury by suppressing its target Sema3 A,implying that the miR-142-3 p/Sema3 A axis may be a new target for preventing keratinocyte injury process.

关 键 词：MiR-142-3p 异常增殖 炎症反应 SEMA3A 银屑病 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]