探讨沉默NOD样受体蛋白3(NLRP3)对银屑病样HaCaT细胞增殖及细胞因子表达的影响。以人角质细胞生长因子(KGF)处理HaCaT细胞,分为KGF组,阴性对照组,NLRP3-RNAi组和对照组。CCK8检测细胞增殖,克隆形成实验检测细胞克隆形成率,流式细胞术检测细胞周期分布,细胞周期蛋白B1(细胞周期蛋白B1)的表达,细胞周期蛋白依赖性激酶2(CDK2),Westernblot检测Ki67和增殖细胞核抗原(PCNA)蛋白,酶联免疫吸附法检测白细胞介素(IL)-17、IL-23、IL-6和肿瘤坏死因子α(TNF-α)水平。与对照组相比,NLRP3mRNA和蛋白的表达,KGF组增殖率和克隆形成率增加,G0/G1期细胞百分比降低,S期细胞百分比增加,CyclinB1、CDK2、Ki67和PCNA蛋白表达增加,IL-17、IL-23、IL-6和TNF-α水平升高。与阴性对照组相比,NLRP3mRNA和蛋白的表达,NLRP3-RNAi组增殖率和克隆形成率降低,G0/G1期细胞百分比增加,S期细胞百分比降低,CyclinB1、CDK2、Ki67和PCNA蛋白的表达均降低,IL-17、IL-23、IL-6和TNF-α水平降低。沉默NLRP3基因可抑制银屑病样HaCaT细胞增殖,阻滞细胞周期,抑制细胞增殖相关蛋白的表达,降低促炎因子的水平。
We aimed to explore the effects of silencing NOD-like receptor protein 3 (NLRP3) on proliferation of psoriasis-like HaCaT cells and expressions of cytokines. HaCaT cells were treated with human keratinocyte growth factor (KGF) and were divided into KGF group, negative control group, NLRP3-RNAi group and control group. Cells proliferation was detected by CCK8, cell clone formation rate was detected by clone formation assay, distribution of cells cycle was detected by flow cytometry, expressions of cyclin B1 (Cyclin B1), cyclin-dependent kinase 2 (CDK2), Ki67 and proliferating cell nuclear antigen (PCNA) proteins were detected by Western blot, and levels of interleukin (IL)-17, IL-23, IL-6 and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay. Compared with control group, expressions of NLRP3 mRNA and protein, proliferation rate and clonal formation rate were increased in KGF group, percentage of cells in G0/G1 phase was decreased, percentage of cells in S phase was increased, expressions of Cyclin B1, CDK2, Ki67 and PCNA proteins were increased, and levels of IL-17, IL-23, IL-6 and TNF-α were increased. Compared with negative control group, expressions of NLRP3 mRNA and protein, proliferation rate and clonal formation rate were decreased in NLRP3-RNAi group, percentage of cells in G0/G1 phase was increased, percentage of cells in S phase was decreased, expressions of Cyclin B1, CDK2, Ki67 and PCNA proteins were decreased, and levels of IL-17, IL-23, IL-6 and TNF-α were decreased. Silencing NLRP3 gene can inhibit the proliferation of psoriasis-like HaCaT cells, arrest cell cycle, inhibit the expressions of cell proliferation-related proteins and reduce levels of pro-inflammatory factors.