伪槐蛇毒部分抗癌和抗菌性能的体外评价

J. Zargan, Majid Mirzaei Nodushan, H. Sobati, Ashkan Haji Noormohammadi, H. Goodarzi, F. Ebrahimi
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引用次数: 1

摘要

背景与目的近年来,从蛇毒中分离出有效分子作为药物治疗一些疑难杂症受到许多生物学研究中心的重视。本研究的目的是测定葡萄假蜡毒的电泳图谱和色谱图谱,并研究其各组分在人肝癌细胞株(HepG2)中的杀菌和抗癌作用。对象与方法采用制备型反相高效液相色谱法对蛇毒进行分离和收集。采用MTT法和MIC法研究了20μg/ml蛋白浓度下各组分对革兰氏阴性菌和革兰氏阳性菌的杀菌活性。然后,在HepG2癌细胞暴露于20和40 μg/ml的蛋白质浓度后,通过MTT还原、中性摄取和彗星试验来测量具有较高抗菌性能的组分的细胞毒作用。结果该毒的SDS-PAGE图谱显示出分子量在13.2 ~ 99.25 kDa范围内的10个主要条带,HPLC分离得到了11个不同的组分。本研究结果表明,与革兰氏阴性菌相比,两个组分(4和8)对革兰氏阳性菌具有显著的抗菌活性。对这两个馏分的抗癌作用的研究也表明,馏分8对HepG2细胞的毒性更大。结论首次报道了波斯角蝰蛇毒液分离部位的杀菌和抗癌作用。这些特性使这些组分成为分离具有抗菌和抗肿瘤活性的分子的有利来源。
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In-Vitro Evaluation of Anticancer and Antibacterial Properties of Pseudocerastes Persicus Snake Venom Fractions
Background and Objectives In recent years the isolation of effective molecules from snake venom as drug to treatment of some incurable diseases is considered by many biological research centers. The aim of this investigation was to determine electrophorotic and chromatographic patterns of Pseudocerastes persicus venom and in-vitro study of bactericidal and anti-cancer properties of its fractions in human hepatocellular liver carcinoma cell line (HepG2). Subjects and Methods The fractions of venom were isolated and collected using preparative RP-HPLC. The Bactericidal activity of fractions in 20μg/ml of protein concentration was investigated toward Gramnegative and Gram-positive bacteria using MTT and MIC assays. Then, the cytotoxic effects of fractions with higher antibacterial properties were measured by MTT reduction, neutral uptake and comet assays following exposure of HepG2 cancer cells to 20 and 40 μg/ml of protein concentration. Results SDS-PAGE pattern of the crude venom revealed 10 major bands with molecular weight ranging from 13.2 to 99.25 kDa and the separation of fractions from the venom by HPLC resulted in collection of 11 different fractions. The results of this study showed that two fractions (4 & 8) have a significant antibacterial activity against Gram-positive as compared to Gram- negative bacteria. The study of the anti-cancer effect of these two fractions also showed that fraction 8 was more toxic in HepG2 cells. Conclusion We for the first time reported bactericidal and anti cancer effects of isolated fractions of Persian horned viper venom. These properties make components of these fractions a favorable source for isolation of molecule (s) with antibacterial and anti tumor activities.
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