ALA-PDT通过HIPPO/YAP信号通路抑制人HSAS1细胞活力和细胞因子分泌

ALA-PDT inhibits human HSAS1 cell viability and cytokine secretion by HIPPO/YAP signaling pathway

目的探讨光动力疗法(ALA-PDT)对人HSAS1成纤维细胞活力和细胞因子分泌的作用及机制。方法ALA-PDT作用HASA1细胞,采用CCK-8法检测细胞活力;ELISA定量试剂盒和Western blot检测TGF-β和VEGF蛋白表达水平的变化;qRT-PCR和Western blot检测YAP、TAZ和LATS1的mRNA和蛋白表达水平的改变。同时,加入YAP抑制剂HY-111249或过表达YAP,观察对ALA-PDT处理HASA1细胞的影响。结果CCK-8法活力实验、ELISA、Western blot和qRT-PCR显示,与Blank组相比,5J ALA-PDT组、10J ALA-PDT组、20J ALA-PDT组细胞活力显著降低(P<0.05),TGF-β和VEGF蛋白表达水平显著降低(P<0.05),YAP和TAZ的mRNA和蛋白表达水平显著下降,LATS1的mRNA和蛋白表达水平显著升高。YAP抑制剂HY-111249可增强ALA-PDT对HSAS1成纤维细胞活力和细胞因子分泌的抑制作用,过表达YAP逆转了ALA-PDT对HSAS1成纤维细胞的抑制作用。结论ALA-PDT可显著抑制人成纤维细胞系HASA1的活力与细胞因子分泌能力,其机制可能与调控Hippo通路关键蛋白YAP和TAZ的表达相关。该研究为ALA联合PDT治疗痤疮疗效提升和改善痤疮的预后提供了一个新的思路和实验理论依据。

5-Aminolevulinic acid-mediated photodynamic therapy(ALA-PDT)is effective in the treatment of acne and however,but the mechanism of ALA-PDT in the treatment of acne remains unclear.The aim of this study was to investigate the role of ALA-PDT in HSAS1 cell proliferation and cytokine secretion and the mechanism.HASA1 cells were treated with ALA-PDT,then the viability of HASA1 cells was detected by CCK-8 assay,the expression levels of TGF-βand VEGF proteins were detected by ELISA quantitative kit and Western blotting,and the mRNA and protein expression levels of YAP,TAZ and LATS1 were detected by qRT-PCR and Western blotting.Meanwhile,YAP inhibitor HY-111249 addition andYAP overexpression were induced to observe their effect on ALA-PDT treatment of HASA1 cells.Data showed that compared with the blank group,ALA-PDT groups(5 J,10 J and 20 J groups)demonstrated lower levels of cell proliferation ability(P<0.05),TGF-βand VEGF protein expression(P<0.05),as well as YAP and TAZ mRNA and protein expression,but higher level of LATS1 mRNA and protein expression.The YAP inhibitor HY-111249 enhanced the inhibitory effect of ALA-PDT on HSAS1 fibroblast viability and cytokine secretion,while overexpression of YAP reversed the inhibitory effect of ALA-PDT on HSAS1 fibroblasts.In conclusion,ALA-PDT can significantly inhibit the viability and cytokine secretion of human fibroblast cell line HASA1,and the mechanism may related to the regulation of YAP and TAZ,which are key proteins of Hippo pathway.This study provides a new idea and experimental theoretical basis for ALA combined with PDT in the treatment of acne.