Nihan Ünübol, İbrahim Çavuş, Tuba Polat, Özgür Kurt, Ahmet Özbilgin, Tanıl Kocagöz
{"title":"抗菌肽及其对滋养利什曼原虫的体外抗利什曼作用","authors":"Nihan Ünübol, İbrahim Çavuş, Tuba Polat, Özgür Kurt, Ahmet Özbilgin, Tanıl Kocagöz","doi":"10.4274/tpd.galenos.2024.48658","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>Antimicrobial resistance is a real threat to humanity. Pentavalent antimonials are reported non-effective in leishmaniasis treatment today, in countries like India. New treatment options have been assessed worldwide lately. Antimicrobial peptides (AMP) are the leading antibiotic candidates due to their large spectrum, fast efficacy, and low resistance risks. Cathelicidins are the AMP with well-documented antimicrobial activities against bacteria, fungi, and protozoa, over their positively charged membranes. Here, we aim to design cathelicidine-like helical peptides (CLHP), and compare their anti-<i>Leishmanial</i> efficacies <i>in vitro</i>, with meglumine antimoniate (MA) on <i>Leishmania tropica</i>.</p><p><strong>Methods: </strong>A total of five study [TN-1-5] and two control (MA and non-drug) groups were formed. Cryopreserved <i>L. tropica</i> isolate was thawed and cultivated in Novy-MacNeal-Nicolle medium and then in RPMI. Five different CLHPs (TN1-5) were diluted in dimethyl sulphoxide. A total of 150 uL of CLHPs and MA were added into the first wells of the test plaques, followed by serial dilutions that revealed doses within 4 and 512 ug/mL. Then, 100 uL of cultures including 1x10<sup>8</sup>/mL of <i>L. tropica</i> promastigotes were added into each well. Viability of promastigotes was checked with XTT, while the parasite count was assessed at 24<sup>th</sup> and 48<sup>th</sup> hours.</p><p><strong>Results: </strong>TN3 was effective at 32 ug/mL. All tested CLHPs exhibited varying degrees of anti-<i>Leishmanial</i> activities, except TN5, even at its highest dose.</p><p><strong>Conclusion: </strong>TN3 showed a particular efficacy against <i>L. tropica</i> <i>in vitro</i>. Further studies including <i>in vivo</i> testing of the candidate's both efficacy and toxicity are essential.</p>","PeriodicalId":34974,"journal":{"name":"Turkiye parazitolojii dergisi","volume":"48 3","pages":"135-141"},"PeriodicalIF":0.0000,"publicationDate":"2024-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Antimicrobial Peptides and Their Anti-<i>Leishmanial</i> Efficacies on <i>Leishmania</i> tropica Promastigotes <i>In vitro</i>.\",\"authors\":\"Nihan Ünübol, İbrahim Çavuş, Tuba Polat, Özgür Kurt, Ahmet Özbilgin, Tanıl Kocagöz\",\"doi\":\"10.4274/tpd.galenos.2024.48658\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>Antimicrobial resistance is a real threat to humanity. Pentavalent antimonials are reported non-effective in leishmaniasis treatment today, in countries like India. New treatment options have been assessed worldwide lately. 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Then, 100 uL of cultures including 1x10<sup>8</sup>/mL of <i>L. tropica</i> promastigotes were added into each well. Viability of promastigotes was checked with XTT, while the parasite count was assessed at 24<sup>th</sup> and 48<sup>th</sup> hours.</p><p><strong>Results: </strong>TN3 was effective at 32 ug/mL. All tested CLHPs exhibited varying degrees of anti-<i>Leishmanial</i> activities, except TN5, even at its highest dose.</p><p><strong>Conclusion: </strong>TN3 showed a particular efficacy against <i>L. tropica</i> <i>in vitro</i>. 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引用次数: 0
摘要
目的:抗菌药耐药性是对人类的真正威胁。据报道,目前在印度等国家,五价抗锑剂对利什曼病治疗无效。最近,全世界都在评估新的治疗方案。抗菌肽(AMP)因其谱广、疗效快、抗药性风险低而成为主要的候选抗生素。柔毛鞘氨醇是一种带正电荷的抗菌肽,对细菌、真菌和原生动物具有良好的抗菌活性。在此,我们旨在设计类猫头鹰螺旋肽(CLHP),并比较它们与巨鲁明抗疟原虫药(MA)在体外对利什曼原虫的抗利什曼病菌效果:方法:共分为五个研究组[TN-1-5]和两个对照组(MA 和非药物)。解冻冷冻的滋养层利什曼原虫分离株,在 Novy-MacNeal-Nicolle 培养基和 RPMI 培养基中培养。用二甲基亚砜稀释五种不同的 CLHP(TN1-5)。将总共 150 uL 的 CLHPs 和 MA 加入测试斑块的第一个孔中,然后进行序列稀释,结果显示剂量在 4 和 512 ug/mL 之间。然后,在每个孔中加入 100 uL 培养物,其中包括 1x108/mL 的 L. tropica 原虫。用 XTT 检验原生体的活力,并在第 24 和 48 小时评估寄生虫数量:结果:TN3 在 32 微克/毫升时有效。除 TN5 外,所有测试的 CLHPs 都表现出不同程度的抗利什曼病菌活性,即使在最高剂量下也是如此:结论:TN3在体外对L.结论:TN3 在体外对 L. tropica 具有特殊疗效,有必要对候选药物进行进一步研究,包括体内疗效和毒性测试。
Antimicrobial Peptides and Their Anti-Leishmanial Efficacies on Leishmania tropica Promastigotes In vitro.
Objective: Antimicrobial resistance is a real threat to humanity. Pentavalent antimonials are reported non-effective in leishmaniasis treatment today, in countries like India. New treatment options have been assessed worldwide lately. Antimicrobial peptides (AMP) are the leading antibiotic candidates due to their large spectrum, fast efficacy, and low resistance risks. Cathelicidins are the AMP with well-documented antimicrobial activities against bacteria, fungi, and protozoa, over their positively charged membranes. Here, we aim to design cathelicidine-like helical peptides (CLHP), and compare their anti-Leishmanial efficacies in vitro, with meglumine antimoniate (MA) on Leishmania tropica.
Methods: A total of five study [TN-1-5] and two control (MA and non-drug) groups were formed. Cryopreserved L. tropica isolate was thawed and cultivated in Novy-MacNeal-Nicolle medium and then in RPMI. Five different CLHPs (TN1-5) were diluted in dimethyl sulphoxide. A total of 150 uL of CLHPs and MA were added into the first wells of the test plaques, followed by serial dilutions that revealed doses within 4 and 512 ug/mL. Then, 100 uL of cultures including 1x108/mL of L. tropica promastigotes were added into each well. Viability of promastigotes was checked with XTT, while the parasite count was assessed at 24th and 48th hours.
Results: TN3 was effective at 32 ug/mL. All tested CLHPs exhibited varying degrees of anti-Leishmanial activities, except TN5, even at its highest dose.
Conclusion: TN3 showed a particular efficacy against L. tropicain vitro. Further studies including in vivo testing of the candidate's both efficacy and toxicity are essential.