Wang Liqun, Zhang Wentao, Cen Ruiyan, Yue Chenda, Xiao Tianzhen, Deng Yumeng, Li Lingfei, Sun Kedai, Lei Xia
Department of Dermatology, Daping Hospital, The Army Medical University, Chongqing, China.
Lasers Surg Med. 2022 Dec;54(10):1309-1320. doi: 10.1002/lsm.23618. Epub 2022 Nov 20.
Photodynamic therapy (PDT) is a promising new approach to promote wound healing and its effectiveness has been demonstrated in both clinical and animal studies. Macrophages are the key cells in wound healing and inflammatory response. However, the mechanism of action of PDT on macrophages in promoting wound healing is still unclear.
In this study, RAW264.7 cells were used. We analyzed the expression levels of macrophage markers arginase 1 (Arg-1), CD206, iNOS, CD86, and inflammatory factors IL-6, TNF-α, and IL-1β by reverse transcription-polymerase chain reaction and Western blot, Milliplex microtubule-associated protein multiplex assay was performed to analyze the expression of inflammatory factors in the supernatant. Live cell Imaging System to observe the dynamic process of macrophage phagocytosis. Western blot was performed to observe the activation of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) and NOD-like receptor protein 3 (NLRP3) inflammasome.
5-Aminolevulinic acid (ALA)-PDT increased the expression of M1 marker iNOS/CD86 and decreased the expression of Arg-1/CD206 in RAW264.7 cells, while, proinflammatory factors IL-6, TNF-α, and IL-1β expression was enhanced and macrophage phagocytosis was increased. We also found that these phenomena were associated with activation of the ERK/MAPK-NLRP3 pathway.
ALA-PDT promotes early inflammatory responses by regulating macrophage M1 polarization through the ERK/MAPK-NLRP3 pathway. It also promotes macrophage phagocytosis.
光动力疗法（PDT）是一种有前景的促进伤口愈合的新方法，其有效性已在临床和动物研究中得到证实。巨噬细胞是伤口愈合和炎症反应中的关键细胞。然而，PDT在促进伤口愈合过程中对巨噬细胞的作用机制仍不清楚。
在本研究中，使用RAW264.7细胞。通过逆转录-聚合酶链反应和蛋白质免疫印迹法分析巨噬细胞标志物精氨酸酶1（Arg-1）、CD206、诱导型一氧化氮合酶（iNOS）、CD86以及炎症因子白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的表达水平，采用Milliplex微管相关蛋白多重检测法分析上清液中炎症因子的表达。利用活细胞成像系统观察巨噬细胞吞噬的动态过程。通过蛋白质免疫印迹法观察细胞外信号调节激酶/丝裂原活化蛋白激酶（ERK/MAPK）和NOD样受体蛋白3（NLRP3）炎性小体的激活情况。
5-氨基酮戊酸（ALA）-PDT增加了RAW264.7细胞中M1标志物iNOS/CD86的表达，降低了Arg-1/CD206的表达，同时促炎因子IL-6、TNF-α和IL-1β的表达增强，巨噬细胞吞噬作用增强。我们还发现这些现象与ERK/MAPK-NLRP3通路的激活有关。
ALA-PDT通过ERK/MAPK-NLRP3通路调节巨噬细胞M1极化，促进早期炎症反应，还促进巨噬细胞吞噬作用。
