You Jianhua, Wang Zhongyun, Jia Xiaoxiao
Department of Dermatology, Zhejiang Provincial Hospital of Dermatology, Huzhou, 313200, China.
Beauty Physiotherapy Department, Zhejiang Provincial Hospital of Dermatology, Huzhou, 313200, China.
Arch Dermatol Res. 2025 Jan 4;317(1):165. doi: 10.1007/s00403-024-03669-8.
Psoriasis is a long-lasting inflammatory skin condition characterized by excessive keratinocyte growth. Recent studies have confirmed abnormal regulation of microRNAs (miRNAs/miRs) in individuals with psoriasis. This study aimed to investigate the function and specific mechanism of action of miR-128a-3p in interleukin-22 (IL-22)-stimulated HaCaT cells. The expression level of miR-128-3p and sirtuin 1 (SIRT1)/hypoxia inducible factor (HIF-1α) was detected using qRT-PCR on patients with psoriasis and IL-22-treated HaCaT cell model. Western blotting was used to detect apoptosis-associated proteins and SIRT1/HIFα pathway protein expression levels. The cell viability was determined using the CCK-8 method. Flow cytometry was performed to detect apoptosis following IL-22 stimulation or transfection. Enzyme-linked immunosorbent assay (ELISA) was used to detect cellular inflammatory factor secretion. The relationship between miR-128-3p and SIRT1 was predicted using the Starbase database and verified using a dual-luciferase reporter gene assay. In patients with psoriasis and IL-22-stimulated HaCaT cells, miR-128-3p and HIF-1α expression levels were elevated and SIRT1 expression was decreased. miR-128-3p directly targeted SIRT1. IL-22 stimulation significantly elevated cell viability, inhibited apoptosis levels and cleaved-caspase3 protein expression, and promoted an inflammatory response in HaCaT cells, which was further promoted by the miR-128-3p mimic. The miR-128-3p inhibitor reduced cell viability, promoted cell apoptosis and cleaved-caspase3 protein expression, and inhibited the inflammatory response in IL-22-induced HaCaT cells; these effects were at least partly reversed by SIRT1-siRNA. miR-128-3p expression is elevated in psoriasis and promotes psoriasis progression by inhibiting SIRT1 expression.
银屑病是一种以角质形成细胞过度生长为特征的慢性炎症性皮肤病。最近的研究证实,银屑病患者体内的微小RNA(miRNA/miR)存在异常调控。本研究旨在探讨miR-128a-3p在白细胞介素-22(IL-22)刺激的HaCaT细胞中的功能及具体作用机制。采用qRT-PCR检测银屑病患者及IL-22处理的HaCaT细胞模型中miR-128-3p和沉默调节蛋白1(SIRT1)/缺氧诱导因子(HIF-1α)的表达水平。采用蛋白质免疫印迹法检测凋亡相关蛋白及SIRT1/HIFα信号通路蛋白表达水平。采用CCK-8法检测细胞活力。采用流式细胞术检测IL-22刺激或转染后的细胞凋亡情况。采用酶联免疫吸附测定(ELISA)检测细胞炎性因子分泌情况。利用Starbase数据库预测miR-128-3p与SIRT1的关系,并通过双荧光素酶报告基因检测进行验证。在银屑病患者及IL-22刺激的HaCaT细胞中,miR-128-3p和HIF-1α表达水平升高,SIRTl表达水平降低。miR-128-3p直接靶向SIRT1。IL-22刺激显著提高了HaCaT细胞的活力,抑制了细胞凋亡水平及caspase3蛋白的表达,并促进了炎性反应,miR-128-3p模拟物进一步增强了这些作用。miR-128-3p抑制剂降低了细胞活力,促进了细胞凋亡及caspase3蛋白的表达,并抑制了IL-22诱导的HaCaT细胞的炎性反应;SIRT1-siRNA至少部分逆转了这些作用。银屑病中miR-128-3p表达升高,并通过抑制SIRT1表达促进银屑病进展。