非洲鲶鱼细菌包膜裂解及多肽的血液毒性研究

Hedmon Okella, C. O. Ajayi, Hilda Ikiriza, Andrew G. Mtewa, Bruhan Kaggwa, J. Aber, C. Kato, P. O. Engeu
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引用次数: 1

摘要

背景:鱼类的皮肤黏液层被赋予了生物制剂,包括抗菌肽(amp),它提供了抵抗病原体的第一道防线。这种多肽既可以抑制细菌生长,也可以完全杀死细菌,因此被认为是传统抗生素的可行替代品,可以解决日益增加的抗菌素耐药性。然而,amp使用的主要障碍之一是其较差的溶血特性。因此,在药物发现的早期阶段,对潜在amp的细菌细胞膜破坏和溶血潜能进行全面评估是至关重要的。目前的研究展示了先前从非洲鲶鱼(Clarias gariepinus)皮肤粘液中分离的抗菌肽的细胞膜破坏和血液相容性。方法:采用15%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对非洲鲶鱼(C. gariepinus)皮肤黏液中分离的抗菌肽进行分析。电导率和碱性磷酸酶测定被用来测量细菌包膜裂解活性作为抗菌肽的经典作用模式。然后用新鲜的兔血细胞进行体外溶血试验。结果:该肽分子量约为5kda,具有破坏细菌胞膜的能力,可引起胞质周围碱性磷酸酶和胞质电解质的显著渗漏。即使在肽提取物最高浓度为100 μg/mL时,新鲜兔血细胞也未见明显溶血现象[3.63%;P>]。[05],这表明它们在正常哺乳动物细胞上是安全的。结论:本研究结果提示加里滨棘猴皮肤黏液中的抗菌肽是潜在安全的抗菌药物先导物来源;然而,仍需要进一步的研究来寻找对宿主细胞安全但对感染细菌有效的最大剂量。
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Bacterial Cell Envelope Lysis and Hemotoxicity of Peptides Previously isolated From African Catfish, Clarias gariepinus
Background: The skin mucus layer of fish is endowed with biologics including, Antimicrobial peptides (AMPs) that offer a first line of defence against pathogens. Such peptides can either inhibit bacterial growth or completely kill the bacteria and hence are regarded as a viable alternative to traditional antibiotics, in addressing the ever-increasing incidences of antimicrobial resistance. However, one of the major hurdles to AMPs use is their poor haemolytic profile. As a result, a thorough evaluation of prospective AMPs’ bacterial cell membrane disruption and hemolytic potentials in the early phases of drug discovery is critical. The current study presented cell membrane destruction as well as hemo-compatibility of antimicrobial peptides previously isolated from skin mucus of African catfish, Clarias gariepinus. Methods: A previously isolated antimicrobial peptide in the skin mucus of African catfish, C. gariepinus were profiled using 15% Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The electrical conductivity and alkaline phosphatase assays were utilised to measure bacterial cell envelope lysis activity as a classical mode of action of the antimicrobial peptides. Afterwards, fresh Rabbit blood cells were then utilised for in vitro hemolytic assay. Results: The peptides were found to be about 5 kDa molecular weight with, ability to damage the bacterial cell envelope causing significant leakage in periplasmic alkaline phosphatase enzyme and cytoplasmic electrolytes. Even at the highest peptide extract concentration of 100 μg/mL, no significant hemolysis was observed on the fresh rabbit blood cells [3.63%;P>.05], signifying their safety on normal mammalian cells. Conclusion: The findings of this study pointed out that antimicrobial peptides in skin mucus of C. gariepinus are potentially safe source of antimicrobial drug leads; however, further studies are still required to search for possibly maximum dose that is safe to host cells but still effective against infecting bacteria.
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