Hyun Jae-Eun, Hwang Cheol-Yong
Department of Veterinary Internal Medicine, College of Veterinary Medicine, Konkuk University, Seoul 05029, Republic of Korea.
Laboratory of Veterinary Dermatology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea.
Vet Sci. 2024 May 23;11(6):235. doi: 10.3390/vetsci11060235.
The direct effects and antimicrobial activity of synthetic antimicrobial peptides (AMPs) obtained from dogs, including cBD, cBD103, and cCath, against wild-type strain PAO1 and canine keratinocytes were analyzed. Antibacterial effects on planktonic bacteria were assessed by determining the minimum bactericidal concentrations (MBCs) of AMPs and by a time-kill assay. Antibiofilm effects were assessed using the microtiter plate assay. We also evaluated the effects of AMPs on cell cytotoxicity and host immune response induced by stimulating canine epidermal keratinocyte progenitor (CPEK) cells with PAO1 and its LPS. cBD, cBD103, and cCath all exhibited dose-dependent antimicrobial and antibiofilm effects. In particular, 25 μg/mL cBD103 showed rapid bactericidal activity within 60 min and inhibited biofilm formation. In addition, pretreatment with cBD103 (25 µg/mL) and cCath (50 µg/mL) 1 h before stimulation significantly reduced the cytotoxicity of the CPEK cells by PAO1 and LPS-induced IL-6 and TNF-a expressions. cBD had little effect on the response to PAO1 and LPS in the cells. These results indicate the therapeutic potential of AMPs in skin infections. However, further studies on the mechanism of action of AMPs in keratinocytes and clinical trials are needed.
分析了从狗身上获得的合成抗菌肽（AMPs），包括cBD、cBD103和cCath，对野生型菌株PAO1和犬角质形成细胞的直接作用及抗菌活性。通过测定AMPs的最低杀菌浓度（MBCs）和进行时间杀菌试验来评估对浮游细菌的抗菌作用。使用微量滴定板试验评估抗生物膜作用。我们还评估了AMPs对用PAO1及其脂多糖刺激犬表皮角质形成细胞祖细胞（CPEK）所诱导的细胞毒性和宿主免疫反应的影响。cBD、cBD103和cCath均表现出剂量依赖性的抗菌和抗生物膜作用。特别是，25μg/mL的cBD103在60分钟内显示出快速杀菌活性并抑制生物膜形成。此外，在刺激前1小时用cBD103（25μg/mL）和cCath（50μg/mL）预处理可显著降低PAO1和脂多糖诱导的CPEK细胞的细胞毒性，以及IL-6和TNF-α的表达。cBD对细胞中PAO1和脂多糖的反应影响很小。这些结果表明AMPs在皮肤感染中的治疗潜力。然而，需要进一步研究AMPs在角质形成细胞中的作用机制并进行临床试验。
