丹皮酚通过STAT3通路抑制IL-17A诱导的角质形成细胞活性和细胞因子分泌

Inhibitory effects of paeonol on keratinocyte viability,cytokines secre⁃tion stimulated by IL-17A via STAT3 signaling pathway

目的:观察丹皮酚对白细胞介素17A(IL-17A)诱导的人角质形成细胞活性、细胞因子分泌及相关信号转导通路的影响。方法:采用IL-17A(200μg/L)刺激体外培养的HaCaT细胞,同时加入不同浓度(200 mg/L和100 mg/L)丹皮酚共同培养24 h。采用CCK-8法测定细胞的活力;CBA法测定Th1/Th2/Th17类炎症细胞因子的变化;ELISA法测定炎症细胞因子IL-23的水平;real-time PCR法测定炎症因子IL-23、IL-6、CXCL2、CXCL8、CCL20和STAT3的mRNA表达;Western blot法测定信号转导通路中STAT3和ERK1/2的蛋白水平。结果:丹皮酚能够显著抑制IL-17A诱导的HaCaT细胞的活力(P<0.05);丹皮酚能够减少IL-6的分泌,同时,能够降低IL-23、CXCL2、CX⁃CL8和CCL20的mRNA表达,并对细胞内STAT3的mRNA和蛋白表达有一定的抑制作用,而对ERK1/2的蛋白水平无显著影响。结论:丹皮酚能够抑制IL-17A诱导的HaCaT细胞的活性和细胞因子的分泌,其作用机制可能与STAT3通路有关。

AIM:To observe the effect of paeonol on interleukin-17A(IL-17A)-induced human keratinocyte viability,cytokine secretion,and related signal transduction pathways.METHODS:In vitro HaCaT cells stimulated by IL-17A(200μg/L)were co-cultured with paeonol(200 mg/L and 100 mg/L)for 24 h.The cell viability was measured by CCK-8 assay.The Th1/Th2/Th17 cytokine(including IL-6,etc.)levels were measured by cytometric bead array assay.The IL-23 level was measured by ELISA.The mRNA expression of IL-23,IL-6,CXCL2,CXCL8,CCL20 and STAT3 was detected by real-time PCR,and Western blot was used to determine the protein levels of STAT3 and ERK1/2.RE⁃SULTS:Paeonol significantly inhibited IL-17A-induced HaCaT cell viability(P<0.05),as well as reduced IL-6 level.Meanwhile,paeonol decreased mRNA levels of IL-23,CXCL2,CXCL8,and CCL20.Paeonol also inhibited the expres⁃sion of STAT3 at mRNA and protein levels.However,no significant effect of paeonol on ERK1/2 protein expression was observed.CONCLUSION:Paeonol inhibits HaCaT cell viability and cytokine secretion induced by IL-17A,and its mechanism might be related to STAT3 singaling pathway.