Cell-free extracts of Propionibacterium acnes stimulate cytokine production through activation of p38 MAPK and Toll-like receptor in SZ95 sebocytes
Cell-free extracts of Propionibacterium acnes stimulate cytokine production through activation of p38 MAPK and Toll-like receptor in SZ95 sebocytes
复制标题
DOI：
10.1016/j.lfs.2015.07.028
复制
发表时间：
2015-10-15
期刊：
影响因子：
6.1
通讯作者：
Hsu, Han-Chi
中科院分区：
文献类型：
--
作者：
Huang, Yu-Chun;Yang, Chao-Hsun;Hsu, Han-Chi
目的：痤疮丙酸杆菌被认为是影响痤疮病变的重要因素。本研究旨在阐明痤疮丙酸杆菌在人皮脂腺细胞中与促炎细胞因子产生相关的潜在信号通路。主要方法：将处于稳定生长期的痤疮丙酸杆菌的无细胞提取物与人永生化的SZ 95皮脂腺细胞共孵育。然后，通过使用定量RT-PCR和ELISA测量mRNA和蛋白质水平来评估无细胞痤疮丙酸杆菌提取物诱导的细胞因子表达。Western blot和Milliplex法检测磷酸化细胞信号蛋白和转录因子的变化。本研究通过shRNA和信号转导通路特异性抑制剂研究了痤疮丙酸杆菌与皮脂腺细胞相互作用的分子机制。主要发现：痤疮丙酸杆菌无细胞提取物显著刺激了SZ 95皮脂腺细胞分泌白细胞介素（IL）-8和IL-6。证明了I κ B-α的降解和I κ B-α、p38丝裂原活化蛋白激酶（MAPK）、CREB和STAT 3的磷酸化增加。定量RT-PCR测量显示，IL-8和Toll样受体2（TLR 2）的基因表达增强痤疮丙酸杆菌的无细胞提取物。此外，NF-κ B抑制剂BMS 345541、p38 MAPK抑制剂SB 203580或抗TLR 2中和抗体阻止无细胞痤疮丙酸杆菌提取物诱导的IL-8分泌。使用shRNA敲低TLR 2对IL-8表达产生类似的抑制作用。STA-21对STAT 3活性的抑制增强了痤疮丙酸杆菌介导的IL-8分泌。意义：痤疮丙酸杆菌无细胞提取物能够激活人SZ 95皮脂腺细胞NF-κ B和p38 MAPK通路，并通过TLR 2依赖性信号上调IL-8的分泌。(C)2015 Elsevier Inc. All rights reserved.
Aims: Propionibacterium acnes has been considered to influence the acne lesions. The present study intended to elucidate the underlying signaling pathways of P. acnes in human sebaceous gland cells relative to the generation of proinflammatory cytokines.Main methods: Cell-free extracts of P. acnes under stationary growth phase were co-incubated with human immortalized SZ95 sebocytes. Then, cell-free P. acnes extracts-induced cytokine expression was evaluated by measuring mRNA and protein levels using quantitative RT-PCR and ELISA. Changes of phosphorylated cell signaling proteins and transcription factors were measured by Western blots and Milliplex assay. The interactive molecular mechanisms of P. acnes and sebocytes were examined through use of shRNA and the specific inhibitors of signaling pathways.Key findings: Cell-free extracts of P. acnes significantly stimulated secretion of interleukin (IL)-8 and IL-6 in SZ95 sebocytes. The degradation of I kappa B-alpha and increased phosphorylation of I kappa B-alpha, p38 mitogen activated protein kinase (MAPK), CREB, and STAT3 were demonstrated. Quantitative RT-PCR measurements revealed that gene expression of IL-8 and Toll-like receptor 2 (TLR2) was enhanced by cell-free extracts of P. acnes. In addition, the NF-kappa B inhibitor BMS345541, p38 MAPK inhibitor SB203580, or anti-TLR2 neutralizing antibody prevented cell-free P. acnes extracts-induced secretion of IL-8. Knockdown of TLR2 using shRNA exerted similar inhibitory effects on IL-8 expression. Moreover, inhibition of STAT3 activity by STA-21 enhanced P. acnes-mediated secretion of IL-8.Significance: Cell-free extracts of P. acties are capable to activate NF-kappa B and p38 MAPK pathways and up-regulate secretion of IL-8 through TLR2-dependent signaling in human SZ95 sebocytes. (C) 2015 Elsevier Inc. All rights reserved.