Efficacy of 5-aminolevulinic acid-based photodynamic therapy against keloid compromised by downregulation of SIRT1-SIRT3-SOD2-mROS dependent autophagy pathway.
Efficacy of 5-aminolevulinic acid-based photodynamic therapy against keloid compromised by downregulation of SIRT1-SIRT3-SOD2-mROS dependent autophagy pathway.
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DOI：
10.1016/j.redox.2018.10.011
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发表时间：
2019-01
期刊：
影响因子：
11.4
通讯作者：
Xu M
中科院分区：
文献类型：
--
作者：
Liu T;Ma X;Ouyang T;Chen H;Xiao Y;Huang Y;Liu J;Xu M
瘢痕疙瘩表现出癌症样的特征，没有自发消退，通常在切除后复发。虽然光动力疗法（PDT）是一种很有前途的治疗方法，但其机制的细节仍有待阐明。在这项研究中，我们研究了基于5-氨基乙酰丙酸（5-ALA）的PDT治疗瘢痕疙瘩的机制。发现5-ALA-PDT诱导超氧阴离子依赖性自噬细胞死亡。应用自噬抑制剂3-甲基腺嘌呤（3-MA）可显著阻止5-ALA-PDT诱导瘢痕疙瘩源性成纤维细胞死亡的作用，但Z-VAK-FMK（凋亡抑制剂）则无此作用。有趣的是，5-ALA-PDT促进了SIRT3蛋白的表达和线粒体超氧化物歧化酶2 （SOD2）的活性，但SIRT1蛋白的表达水平降低。SOD2是降低线粒体ROS （mROS）水平的关键酶，SIRT3介导的SOD2去乙酰化可调节SOD2酶活性。然后我们用免疫沉淀法检测SOD2乙酰化水平，发现5-ALA-PDT显著提高了SOD2乙酰化水平。为了证实SIRT3调控SOD2去乙酰化，我们使用了3-TYP （SIRT3抑制剂）。发现与对照组或5-ALA-PDT组相比，3-TYP对SIRT3的抑制显著提高了SOD2乙酰化水平。为了探索SIRT1和SIRT3之间的联系，我们用SIRT1抑制剂EX527或SIRT1激活剂SRT1720处理细胞，EX527增加了SIRT3蛋白水平，而SRT1720在5-ALA-PDT存在或不存在时表现出相反的作用。此外，sirt1抑制的细胞对5-ALA-PDT的抗性更强，ROS积累减少。这些结果可能表明，5-ALA-PDT诱导SIRT1蛋白水平降低，从而促进SIRT3提高SOD2活性的作用，从而降低mROS水平，进而损害5-ALA-PDT诱导的自噬细胞死亡。
Keloids exhibit cancer-like properties without spontaneous regression and usually recur post excision. Although photodynamic therapy (PDT) is a promising treatment, details of the mechanisms remain to be elucidated. In this study, we investigated mechanisms involved in 5-Aminolevulinic Acid (5-ALA)–based PDT against keloid. Found that 5-ALA-PDT induced superoxide anion-dependent autophagic cell death. Application of autophagy inhibitor 3-Methyladenine (3-MA) significantly prevented the effect that 5-ALA-PDT induced keloid–derived fibroblasts death, but Z-VAK-FMK (apoptotic inhibitor) did not. Interestingly, 5-ALA-PDT promoted the SIRT3 protein expression and the activity of mitochondrial superoxide dismutase 2 (SOD2), but SIRT1 protein expression level was decreased. SOD2 as a key enzyme can decrease mitochondrial ROS (mROS) level, Deacetylation of SOD2 by SIRT3 regulates SOD2 enzymatic activity has been identified. Then we explored SOD2 acetylation level with immunoprecipitation, found that 5-ALA-PDT significantly increased the acetylation levels of SOD2. In order to confirm deacetylation of SOD2 regulated by SIRT3, 3-TYP (SIRT3 inhibitor) was used. Found that inhibition of SIRT3 by 3-TYP significantly increased the level of SOD2 acetylation level compared with control group or 5-ALA-PDT group. To explore the connection of SIRT1 and SIRT3, cells were treated with EX527(SIRT1 inhibitor) or SRT1720 (SIRT1 activator), and EX527 increased SIRT3 protein level, however, SRT1720 displayed the opposite effect in the present or absence of 5-ALA-PDT. Moreover SIRT1-inhibited cells are more resistant to 5-ALA-PDT and showing decreased ROS accumulation. These results may demonstrate that 5-ALA-PDT induced SIRT1 protein level decreased, which promoted the effect of SIRT3 increased activity of SOD2 that can reduce mROS level, and then compromised 5-ALA-PDT induced autophagic cell death.
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影响因子：
16.2
作者：
Herskovits AZ;Guarente L
通讯作者：
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影响因子：
29
作者：
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通讯作者：
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影响因子：
10.3
作者：
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影响因子：
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作者：
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Zhou Z
影响因子：
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作者：
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