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植物乳杆菌LR/14产生的非溶血性、非免疫原性、无毒抗菌肽(AMPs LR14)的抗疟活性评估。

Assessment of anti-plasmodial activity of non-hemolytic, non-immunogenic, non-toxic antimicrobial peptides (AMPs LR14) produced by Lactobacillus plantarum LR/14.

作者信息

Gupta Ruchi, Rajendran Vinoth, Ghosh Prahlad C, Srivastava Sheela

机构信息

Department of Genetics, University of Delhi South Campus, New Delhi, 110021, India.

出版信息

Drugs R D. 2014 Jun;14(2):95-103. doi: 10.1007/s40268-014-0043-y.

Abstract

BACKGROUND AND OBJECTIVES

Lactobacillus plantarum strains are known to exhibit an antimicrobial property against bacteria and fungi. In the present investigation, AMPs LR14, antimicrobial peptides produced by L. plantarum strain LR/14, were tested against a protozoan system, Plasmodium falciparum and its non-toxic nature was envisaged on a mammalian system.

METHODS

Human erythrocytes infected with chloroquine-sensitive and -resistant strains of P. falciparum were treated with purified AMPs LR14. The loss in cell viability was assessed by monitoring the incorporation of [(3)H]-hypoxanthine in the nucleic acid of the parasite. The hemolytic activity of AMPs LR14 was monitored at different concentrations and the investigations into the in vivo toxicity of AMPs LR14 were carried out on a mammalian system (Wistar rat). The level of toxicity in the tissues was visualized by histopathological studies conducted on the liver and kidney of the test and control rats. A study was also undertaken to see the production of antibodies in an animal (rabbit) after it was immunized with AMPs LR14.

RESULTS

A loss in cell viability was observed in both test strains of P. falciparum. However, the dose required for inhibition of the chloroquine-resistant strain was ~2 times the dose required for the chloroquine-sensitive strain. At these concentrations, no hemolysis of human erythrocytes was observed. The studies conducted on in vivo toxicity of AMPs LR14 suggest that the lethal dose (LD50) is beyond 1,000 mg/kg body weight, suggesting its safe use against microbes and protozoans. Antibodies were also not detected against these peptides, indicating a non-immunogenic nature.

CONCLUSION

The data indicate that AMPs LR14 are non-toxic, potent anti-plasmodial peptides causing growth inhibition of P. falciparum without causing hemolysis. These results pave the way for the development of bioactive peptides as therapeutics.

摘要

背景与目的

已知植物乳杆菌菌株对细菌和真菌具有抗菌特性。在本研究中,对植物乳杆菌菌株LR/14产生的抗菌肽AMPs LR14针对疟原虫系统恶性疟原虫进行了测试,并设想了其在哺乳动物系统中的无毒性质。

方法

用纯化的AMPs LR14处理感染了氯喹敏感和耐药恶性疟原虫菌株的人红细胞。通过监测[(3)H] - 次黄嘌呤掺入寄生虫核酸中的情况来评估细胞活力的丧失。在不同浓度下监测AMPs LR14的溶血活性,并在哺乳动物系统(Wistar大鼠)上对AMPs LR14的体内毒性进行研究。通过对试验组和对照组大鼠的肝脏和肾脏进行组织病理学研究来观察组织中的毒性水平。还进行了一项研究,观察用AMPs LR14免疫动物(兔子)后抗体的产生情况。

结果

在两种恶性疟原虫测试菌株中均观察到细胞活力丧失。然而,抑制氯喹耐药菌株所需的剂量约为抑制氯喹敏感菌株所需剂量的2倍。在这些浓度下,未观察到对人红细胞的溶血现象。对AMPs LR14体内毒性的研究表明,致死剂量(LD50)超过1000mg/kg体重,表明其对微生物和原生动物的安全使用。也未检测到针对这些肽的抗体,表明其无免疫原性。

结论

数据表明AMPs LR14是无毒、有效的抗疟原虫肽,可抑制恶性疟原虫生长而不引起溶血。这些结果为生物活性肽作为治疗药物的开发铺平了道路。

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