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首页> 外文期刊>Photodiagnosis and Photodynamic Therapy >Pyropheophorbide-a methyl ester-mediated photodynamic therapy induces apoptosis and inhibits LPS-induced inflammation in RAW264.7 macrophages
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Pyropheophorbide-a methyl ester-mediated photodynamic therapy induces apoptosis and inhibits LPS-induced inflammation in RAW264.7 macrophages

机译：焦脱镁叶绿酸-甲酯介导的光动力疗法诱导RAW264.7巨噬细胞凋亡并抑制LPS诱导的炎症

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Background: This study aimed to determine the effect of pyropheophorbide-a methyl ester (MPPa)-mediated photodynamic therapy (MPPa-PDT) on the apoptosis and inflammation of murine macrophage RAW264.7 cells.Methods: Uptake and subcellular localization of MPPa was detected by flow cytometry and confocal fluorescence microscope. Cell viability was assessed by CCK-8; ROS levels were assessed by DCFH-DA. Cell apoptosis was measured by flow cytometry and Hoechst 33342 staining, whereas mitochondrial membrane potential was detected by JC-1 staining. Secretion of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-6 (IL-6) was determined using ELISA kits. Caspase-3, cleaved caspase-3, procaspase-9, cleaved caspase-9, PARP, cleaved PARP, Bcl-2, Bax, NF-kappa B p-p65, p-IKK alpha/beta, and p-ltcBa were measured by western blotting. Nuclear factor kappa B (NF-kappa B)-p65 nuclear translocation was observed by immunofluorescence.Results: MPPa -PDT influenced cell viability in a light dose-dependent manner. It induced ROS formation and RAW264.7 cell apoptosis. It also increased the expression of cleaved caspase-3, cleaved caspase-9, cleaved PARP and Bax, decreased the expression of Bcl-2. While TNF-alpha, IL-1 beta, and lL-6 increased in LPS group (model of inflammation), it deceased in LPS-MPPa-PDT group. NF-kappa B p-p65, p-IKK alpha/beta, and p-I kappa B alpha had higher expression in LPS group while that reduced in LPS-MPPa-PDT group. Simultaneously, MPPa-PDT inhibited nuclear translocation of NF-kappa B-p65 caused by LPS.Conclusions: MPPa-PDT can induce apoptosis and attenuate inflammation in mouse RAW264.7 macrophages, thereby suggesting a promising therapy for atherosclerosis.
机译：背景：本研究旨在确定焦脱镁叶绿酸一甲酯（MPPa）介导的光动力疗法（MPPa-PDT）对鼠巨噬细胞RAW264.7细胞凋亡和炎症的影响。方法：检测MPPa的摄取和亚细胞定位通过流式细胞仪和共聚焦荧光显微镜。通过CCK-8评估细胞活力； ROS水平通过DCFH-DA评估。通过流式细胞术和Hoechst 33342染色测量细胞凋亡，而通过JC-1染色检测线粒体膜电位。使用ELISA试剂盒确定了肿瘤坏死因子α（TNF-alpha），白介素1 beta（IL-1 beta）和白介素6（IL-6）的分泌。测量了caspase-3，裂解的caspase-3，procaspase-9，裂解的caspase-9，PARP，裂解的PARP，Bcl-2，Bax，NF-κB p-p65，p-IKK alpha / beta和p-ltcBa通过蛋白质印迹。免疫荧光观察到核因子κB（NF-κB）-p65核易位。结果：MPPa -PDT以光剂量依赖性方式影响细胞活力。它诱导ROS形成和RAW264.7细胞凋亡。它也增加了裂解的caspase-3，裂解的caspase-9，PARP和Bax的表达，降低了Bcl-2的表达。 LPS组（炎症模型）的TNF-α，IL-1β和1L-6升高，而LPS-MPPa-PDT组则降低。 NF-κB p-p65，p-IKK alpha / beta和p-I kappa B alpha在LPS组中较高表达，而在LPS-MPPa-PDT组中较低。同时，MPPa-PDT抑制LPS引起的NF-κB-p65的核易位。结论：MPPa-PDT可以诱导小鼠RAW264.7巨噬细胞凋亡并减轻炎症，从而为动脉粥样硬化的治疗提供了有希望的方法。

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