Department of Biotechnology and Bioinformatics, JSS Academy of Higher Education and Research, Mysuru 570 015, India.
Department of Microbiology, JSS Academy of Higher Education and Research, Mysuru 570 015, India.
Molecules. 2023 Jan 22;28(3):1106. doi: 10.3390/molecules28031106.
Bacterial infections are one of the leading causes of morbidity, mortality, and healthcare complications in patients. Leptospirosis is found to be the most prevalent, re-emergent, and neglected tropical zoonotic disease worldwide. The adaptation to various environmental conditions has made acquire a large genome (~4.6 Mb) and a complex outer membrane, making it unique among bacteria that mimic the symptoms of jaundice and hemorrhage. Sph2 is another important virulence factor that enhances hemolytic sphingomyelinase-capable of moving inside mitochondria-which increases the ROS level and decreases the mitochondrial membrane potential, thereby leading to cell apoptosis. In the present study, 25 suspected bovine serum samples were subjected to the Microscopic Agglutination Test (MAT) across the Mysuru region. Different samples, such as urine, serum, and aborted materials from the confirmed MAT-positive animals, were used for isolation and genomic detection by conventional PCR targeting virulence gene, Lipl32, using specific primers. Further, in vitro and in silico studies were performed on isolated cultures to assess the anti-leptospiral, anti-hemolytic, and sphingomyelinase enzyme inhibition using novel pseudopeptides. The microdilution technique (MDT) and dark field microscope (DFM) assays revealed that at a concentration of 62.5 μg/mL, the pseudopeptide inhibited 100% of the growth of spp., suggesting its efficiency in the treatment of leptospirosis. The flow cytometry analyses show the potency of the pseudopeptide against sphingomyelinase enzymes using human umbilical vein endothelial cells (HUVECs). Thus, the present study demonstrated the efficacy of the pseudopeptide in the inhibition of the growth of , and therefore, this can be used as an alternative drug for the treatment of leptospirosis.
细菌感染是导致患者发病、死亡和医疗并发症的主要原因之一。钩端螺旋体病是全球最普遍、重新出现和被忽视的热带动物源性传染病。它适应各种环境条件的能力使其获得了一个庞大的基因组（~4.6 Mb）和一个复杂的外膜，使其在模仿黄疸和出血症状的细菌中独一无二。Sph2 是另一个重要的毒力因子，它增强了溶血神经鞘磷脂酶的能力，使其能够进入线粒体——这会增加 ROS 水平并降低线粒体膜电位，从而导致细胞凋亡。在本研究中，对迈索尔地区的 25 份疑似牛血清样本进行了显微镜凝集试验（MAT）。使用针对毒力基因 Lipl32 的常规 PCR 从确诊的 MAT 阳性动物的不同样本（如尿液、血清和流产材料）进行分离和基因组检测，使用的是特定引物。此外，对分离培养物进行了体外和计算机模拟研究，使用新型拟肽评估其抗钩端螺旋体、抗溶血和神经鞘磷脂酶抑制作用。微量稀释技术（MDT）和暗场显微镜（DFM）检测结果表明，在 62.5 μg/mL 的浓度下，拟肽抑制了 spp. 的 100%生长，表明其在治疗钩端螺旋体病方面的效率。流式细胞术分析表明，拟肽对人脐静脉内皮细胞（HUVECs）中的神经鞘磷脂酶具有强大的抑制作用。因此，本研究证明了拟肽抑制 生长的功效，因此可作为治疗钩端螺旋体病的替代药物。
