{"title":"针对白色念珠菌细胞凋亡的人类抗菌肽的抗真菌活性。","authors":"Siham Shaban, Mrudula Patel, Aijaz Ahmad","doi":"10.1099/jmm.0.001835","DOIUrl":null,"url":null,"abstract":"<p><p><b>Introduction.</b> Innovative antifungal therapies are of crucial importance to combat the potentially life-threatening infections linked to the multidrug-resistant fungal pathogen <i>Candida auris</i>. Induction of regulated cell death, apoptosis, could provide an outline for future therapeutics. Human antimicrobial peptides (AMPs), well-known antifungal compounds, have shown the ability to induce apoptosis in pathogenic fungi.<b>Hypothesis/Gap Statement</b> <b>.</b> Although it is known that AMPs possess antifungal activity against <i>C. auris</i>, their ability to induce apoptosis requires further investigations.<b>Aim.</b> This study evaluated the effects of AMPs on the induction of apoptosis in <i>C. auris</i>.<b>Methods.</b> Human neutrophil peptide-1 (HNP-1), human β-Defensins-3 (hBD-3) and human salivary histatin 5 (His 5) were assessed against two clinical <i>C. auris</i> isolates. Apoptosis hallmarks were examined using FITC-Annexin V/PI double labelling assay and terminal deoxynucleotidyl transferase deoxynucleotidyl transferase nick-end labelling (TUNEL) to detect phosphatidylserine externalization and DNA fragmentation, respectively. Then, several intracellular triggers were studied using JC-10 staining, spectrophotometric assay and 2',7'-dichlorofluorescin diacetate staining to measure the mitochondrial membrane potential, cytochrome-c release and reactive oxygen species (ROS) production, respectively.<b>Results and conclusion.</b> FITC-Annexin V/PI staining and TUNEL analysis revealed that exposure of <i>C. auris</i> cells to HNP-1 and hBD-3 triggered both early and late apoptosis, while His 5 caused significant necrosis. Furthermore, HNP-1 and hBD-3 induced significant mitochondrial membrane depolarization, which resulted in substantial cytochrome c release. In contrast to His 5, which showed minimal mitochondrial depolarization and no cytochrome c release. At last, all peptides significantly increased ROS production, which is related to both types of cell death. Therefore, these peptides represent promising and effective antifungal agents for treating invasive infections caused by multidrug-resistant <i>C. auris.</i></p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 5","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Antifungal activity of human antimicrobial peptides targeting apoptosis in <i>Candida auris</i>.\",\"authors\":\"Siham Shaban, Mrudula Patel, Aijaz Ahmad\",\"doi\":\"10.1099/jmm.0.001835\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Introduction.</b> Innovative antifungal therapies are of crucial importance to combat the potentially life-threatening infections linked to the multidrug-resistant fungal pathogen <i>Candida auris</i>. Induction of regulated cell death, apoptosis, could provide an outline for future therapeutics. Human antimicrobial peptides (AMPs), well-known antifungal compounds, have shown the ability to induce apoptosis in pathogenic fungi.<b>Hypothesis/Gap Statement</b> <b>.</b> Although it is known that AMPs possess antifungal activity against <i>C. auris</i>, their ability to induce apoptosis requires further investigations.<b>Aim.</b> This study evaluated the effects of AMPs on the induction of apoptosis in <i>C. auris</i>.<b>Methods.</b> Human neutrophil peptide-1 (HNP-1), human β-Defensins-3 (hBD-3) and human salivary histatin 5 (His 5) were assessed against two clinical <i>C. auris</i> isolates. Apoptosis hallmarks were examined using FITC-Annexin V/PI double labelling assay and terminal deoxynucleotidyl transferase deoxynucleotidyl transferase nick-end labelling (TUNEL) to detect phosphatidylserine externalization and DNA fragmentation, respectively. Then, several intracellular triggers were studied using JC-10 staining, spectrophotometric assay and 2',7'-dichlorofluorescin diacetate staining to measure the mitochondrial membrane potential, cytochrome-c release and reactive oxygen species (ROS) production, respectively.<b>Results and conclusion.</b> FITC-Annexin V/PI staining and TUNEL analysis revealed that exposure of <i>C. auris</i> cells to HNP-1 and hBD-3 triggered both early and late apoptosis, while His 5 caused significant necrosis. Furthermore, HNP-1 and hBD-3 induced significant mitochondrial membrane depolarization, which resulted in substantial cytochrome c release. In contrast to His 5, which showed minimal mitochondrial depolarization and no cytochrome c release. At last, all peptides significantly increased ROS production, which is related to both types of cell death. Therefore, these peptides represent promising and effective antifungal agents for treating invasive infections caused by multidrug-resistant <i>C. auris.</i></p>\",\"PeriodicalId\":94093,\"journal\":{\"name\":\"Journal of medical microbiology\",\"volume\":\"73 5\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of medical microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1099/jmm.0.001835\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of medical microbiology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1099/jmm.0.001835","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
导言。创新的抗真菌疗法对于抗击由具有多重耐药性的真菌病原体白色念珠菌引起的可能危及生命的感染至关重要。诱导调节性细胞死亡(细胞凋亡)可以为未来的疗法提供一个轮廓。人类抗菌肽(AMPs)是著名的抗真菌化合物,已显示出诱导病原真菌细胞凋亡的能力。尽管已知 AMPs 具有抗真菌活性,但其诱导细胞凋亡的能力还需要进一步研究。本研究评估了 AMPs 对诱导 C. auris 细胞凋亡的影响。评估了人中性粒细胞肽-1(HNP-1)、人β-防御素-3(hBD-3)和人唾液组蛋白5(His 5)对两种临床阴道杆菌分离物的作用。使用 FITC-Annexin V/PI 双标记检测法和末端脱氧核苷酸转移酶缺刻端标记法(TUNEL)分别检测磷脂酰丝氨酸外化和 DNA 断裂,以检查细胞凋亡标志。然后,使用JC-10染色法、分光光度法和2',7'-二氯荧光素二乙酸酯染色法分别研究了线粒体膜电位、细胞色素-c释放和活性氧(ROS)产生的几种细胞内触发因素。FITC-Annexin V/PI染色和TUNEL分析表明,C. auris细胞暴露于HNP-1和hBD-3会引发早期和晚期细胞凋亡,而His 5则会导致细胞大量坏死。此外,HNP-1 和 hBD-3 还诱导线粒体膜显著去极化,导致大量细胞色素 c 释放。相比之下,His 5 的线粒体去极化程度很小,也没有细胞色素 c 释放。最后,所有肽都能显著增加 ROS 的产生,而 ROS 的产生与两种细胞死亡类型都有关系。因此,这些多肽是治疗耐多药球孢子菌引起的侵袭性感染的有效抗真菌剂。
Antifungal activity of human antimicrobial peptides targeting apoptosis in Candida auris.
Introduction. Innovative antifungal therapies are of crucial importance to combat the potentially life-threatening infections linked to the multidrug-resistant fungal pathogen Candida auris. Induction of regulated cell death, apoptosis, could provide an outline for future therapeutics. Human antimicrobial peptides (AMPs), well-known antifungal compounds, have shown the ability to induce apoptosis in pathogenic fungi.Hypothesis/Gap Statement. Although it is known that AMPs possess antifungal activity against C. auris, their ability to induce apoptosis requires further investigations.Aim. This study evaluated the effects of AMPs on the induction of apoptosis in C. auris.Methods. Human neutrophil peptide-1 (HNP-1), human β-Defensins-3 (hBD-3) and human salivary histatin 5 (His 5) were assessed against two clinical C. auris isolates. Apoptosis hallmarks were examined using FITC-Annexin V/PI double labelling assay and terminal deoxynucleotidyl transferase deoxynucleotidyl transferase nick-end labelling (TUNEL) to detect phosphatidylserine externalization and DNA fragmentation, respectively. Then, several intracellular triggers were studied using JC-10 staining, spectrophotometric assay and 2',7'-dichlorofluorescin diacetate staining to measure the mitochondrial membrane potential, cytochrome-c release and reactive oxygen species (ROS) production, respectively.Results and conclusion. FITC-Annexin V/PI staining and TUNEL analysis revealed that exposure of C. auris cells to HNP-1 and hBD-3 triggered both early and late apoptosis, while His 5 caused significant necrosis. Furthermore, HNP-1 and hBD-3 induced significant mitochondrial membrane depolarization, which resulted in substantial cytochrome c release. In contrast to His 5, which showed minimal mitochondrial depolarization and no cytochrome c release. At last, all peptides significantly increased ROS production, which is related to both types of cell death. Therefore, these peptides represent promising and effective antifungal agents for treating invasive infections caused by multidrug-resistant C. auris.