Application of bacteriophage φPaP11-13 attenuates rat Cutibacterium acnes infection lesions by promoting keratinocytes apoptosis via inhibiting PI3K/Akt pathway
Application of bacteriophage φPaP11-13 attenuates rat Cutibacterium acnes infection lesions by promoting keratinocytes apoptosis via inhibiting PI3K/Akt pathway
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DOI：
10.1128/spectrum.02838-23
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发表时间：
2024-01
影响因子：
3.7
通讯作者：
Yuanyuan Liu;Ni Zhen;Danxi Liao;Jiahui Niu;Ruolan Liu;Zijiao Li;Zeyuan Lei;Zichen Yang
中科院分区：
文献类型：
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作者：
Yuanyuan Liu;Ni Zhen;Danxi Liao;Jiahui Niu;Ruolan Liu;Zijiao Li;Zeyuan Lei;Zichen Yang
摘要寻常痤疮是由耐药痤疮皮肤杆菌（Cutibacterium acnes，C。痤疮）感染难以常规治疗。噬菌体已被认为是一种潜在的解决方案，但对噬菌体治疗机制的研究还不够。本研究探讨了噬菌体φ PaP11 - 13对C.痤疮诱导的炎症大鼠模型。我们发现感染C.痤疮具有较高的平均耳厚度，苏木精-伊红（HE）染色显示的炎性细胞更丰富，IF染色显示的TUNEL（TdT介导的dUTP缺口末端标记）阳性角质形成细胞更少。免疫组化、Western blot和实时荧光定量PCR（qRT-PCR）检测感染C.痤疮，这是减少后应用噬菌体φ PaP11 - 13。通过应用IGF-1抗体，证明C.痤疮引起的炎症反应与IGF-1的表达有关。WB和qRT-PCR结果显示，C.痤疮感染。随后，显示了在应用噬菌体φ PaP11 - 13后，PI3K/Akt途径对BAD介导的凋亡途径的激活被减轻。IGF-1r抑制剂、Pan-PI3K抑制剂和Akt抑制剂可逆转C.痤疮和噬菌体φ PaP 11 - 13。本研究表明，噬菌体φ PaP11 - 13对寻常痤疮的减毒作用的关键机制之一是裂解C.通过PI3K/Akt信号通路调节角质形成细胞凋亡。痤疮皮肤杆菌感染诱发的寻常痤疮可引起严重的生理和心理预后。然而，抗生素的过度使用会产生耐药性，给治疗痤疮皮肤杆菌带来挑战。噬菌体目前已被证明对MDR（多重耐药）痤疮皮肤杆菌有效，但对噬菌体治疗缺乏了解。本研究通过裂解痤疮皮肤杆菌促进痤疮皮损中过量角质形成细胞的细胞死亡，证明了一种新的治疗寻常痤疮的方法。然而，这种细胞周期的调节尚未被证明是直接介导的。噬菌体-细菌-宿主三元关系的提示激发了未来噬菌体治疗研究的巨大兴趣。痤疮皮肤杆菌感染引起的寻常痤疮可引起严重的生理和心理预后。然而，抗生素的过度使用会产生耐药性，给治疗痤疮皮肤杆菌带来挑战。噬菌体目前已被证明对MDR（多重耐药）痤疮皮肤杆菌有效，但对噬菌体治疗缺乏了解。本研究通过裂解痤疮皮肤杆菌促进痤疮皮损中过量角质形成细胞的细胞死亡，证明了一种新的治疗寻常痤疮的方法。然而，这种细胞周期的调节尚未被证明是直接介导的。噬菌体-细菌-宿主三元关系的提示激发了未来噬菌体治疗研究的巨大兴趣。
ABSTRACT Acne vulgaris caused by antibiotic-resistant Cutibacterium acnes (C. acnes) infection is difficult to treat conventionally. Phages have been suggested as a potential solution, but research on the mechanism of phage treatment is inadequate. This research investigates the underlying molecular mechanisms of phage φPaP11-13 attenuating C. acnes-induced inflammation in rat models. We found that rats infected with C. acnes had higher average ear thickness, greater enrichment of inflammatory cells as shown by hematoxylin–eosin (HE) staining, and fewer TUNEL (TdT-mediated dUTP Nick-End Labeling)-positive keratinocytes visualized by IF staining. Moreover, an increase of IGF-1 and IGF-1 receptor (IGF-1r) was detected using the immunohistochemical (IHC) staining method, Western blot (WB), and quantitative real-time PCR (qRT-PCR) when infected with C. acnes, which was decreased after the application of phage φPaP11-13. By applying the IGF-1 antibody, it was demonstrated that the severity of C. acnes-induced inflammation was relevant to the expression of IGF-1. Through WB and qRT-PCR, activation of the PI3K/Akt pathway and a down-regulation of the BAD-mediated apoptosis pathway were discovered after C. acnes infection. Subsequently, it was shown that the activation of the PI3K/Akt pathway against BAD-mediated apoptosis pathway was alleviated after applying phage φPaP11-13. Furthermore, applying the IGF-1r inhibitor, Pan-PI3K inhibitor, and Akt inhibitor reversed the changing trends of BAD induced by C. acnes and phage φPaP11-13. This study demonstrates that one of the critical mechanisms underlying the attenuation of acne vulgaris by phage φPaP11-13 is lysing C. acnes and regulating keratinocyte apoptosis via the PI3K/Akt signaling pathway. IMPORTANCE Cutibacterium acnes infection-induced acne vulgaris may cause severe physical and psychological prognosis. However, the overuse of antibiotics develops drug resistance, bringing challenges in treating Cutibacterium acnes. Bacteriophages are currently proven effective in MDR (multiple drug-resistant) Cutibacterium acnes, but there is a significant lack of understanding of phage therapy. This study demonstrated a novel way of curing acne vulgaris by using phages through promoting cell death of excessive keratinocytes in acne lesions by lysing Cutibacterium acnes. However, the regulation of this cell cycle has not been proven to be directly mediated by phages. The hint of ternary relation among "phage–bacteria–host" inspires huge interest in future phage therapy studies. Cutibacterium acnes infection-induced acne vulgaris may cause severe physical and psychological prognosis. However, the overuse of antibiotics develops drug resistance, bringing challenges in treating Cutibacterium acnes. Bacteriophages are currently proven effective in MDR (multiple drug-resistant) Cutibacterium acnes, but there is a significant lack of understanding of phage therapy. This study demonstrated a novel way of curing acne vulgaris by using phages through promoting cell death of excessive keratinocytes in acne lesions by lysing Cutibacterium acnes. However, the regulation of this cell cycle has not been proven to be directly mediated by phages. The hint of ternary relation among "phage–bacteria–host" inspires huge interest in future phage therapy studies.