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蜜蜂毒液(意大利蜜蜂)含有抗凝因子并延长凝血时间。

Honey Bee Venom (Apis mellifera) Contains Anticoagulation Factors and Increases the Blood-clotting Time.

作者信息

Zolfagharian Hossein, Mohajeri Mohammad, Babaie Mahdi

机构信息

Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, Iran.

Department of Toxicology, School of Pharmacy, Shahreza Branch, Islamic Azad University, Shahreza, Iran.

出版信息

J Pharmacopuncture. 2015 Dec;18(4):7-11. doi: 10.3831/KPI.2015.18.031.

Abstract

OBJECTIVES

Bee venom (BV) is a complex mixture of proteins and contains proteins such as phospholipase and melittin, which have an effect on blood clotting and blood clots. The mechanism of action of honey bee venom (HBV, Apis mellifera) on human plasma proteins and its anti-thrombotic effect were studied. The purpose of this study was to investigate the anti-coagulation effect of BV and its effects on blood coagulation and purification.

METHODS

Crude venom obtained from Apis mellifera was selected. The anti-coagulation factor of the crude venom from this species was purified by using gel filtration chromatography (sephadex G-50), and the molecular weights of the anti-coagulants in this venom estimated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Blood samples were obtained from 10 rabbits, and the prothrombin time (PT) and the partial thromboplastin time (PTT) tests were conducted. The approximate lethal dose (LD) values of BV were determined.

RESULTS

Crude BV increased the blood clotting time. For BV concentrations from 1 to 4 mg/mL, clotting was not observed even at more than 300 seconds, standard deviations (SDs) = ± 0.71; however, clotting was observed in the control group 13.8 s, SDs = ± 0.52. Thus, BV can be considered as containing anti-coagulation factors. Crude BV is composed 4 protein bands with molecular weights of 3, 15, 20 and 41 kilodalton (kDa), respectively. The LD50 of the crude BV was found to be 177.8 μg/mouse.

CONCLUSION

BV contains anti-coagulation factors. The fraction extracted from the Iranian bees contains proteins that are similar to anti-coagulation proteins, such as phospholipase A2 (PLA2) and melittin, and that can increase the blood clotting times in vitro.

摘要

目的

蜂毒(BV)是一种蛋白质的复杂混合物,含有磷脂酶和蜂毒肽等对血液凝固和血栓有影响的蛋白质。研究了蜜蜂毒液(HBV,意大利蜜蜂)对人血浆蛋白的作用机制及其抗血栓形成作用。本研究的目的是探讨BV的抗凝作用及其对血液凝固和纯化的影响。

方法

选用从意大利蜜蜂获得的粗毒液。使用凝胶过滤色谱法(葡聚糖G - 50)纯化该物种粗毒液的抗凝因子,并使用十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)估计该毒液中抗凝剂的分子量。从10只兔子采集血样,并进行凝血酶原时间(PT)和部分凝血活酶时间(PTT)测试。测定BV的近似致死剂量(LD)值。

结果

粗BV延长了血液凝固时间。对于浓度为1至4mg/mL的BV,即使超过300秒也未观察到凝血,标准差(SDs)=±0.71;然而,对照组在13.8秒观察到凝血,SDs =±0.52。因此,BV可被认为含有抗凝因子。粗BV由4条分别具有3、15、20和41千道尔顿(kDa)分子量的蛋白带组成。发现粗BV的LD50为177.8μg/小鼠。

结论

BV含有抗凝因子。从伊朗蜜蜂中提取的组分含有与抗凝蛋白如磷脂酶A2(PLA2)和蜂毒肽相似的蛋白质,并且在体外可延长血液凝固时间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2d9/4797586/dbd6c983f431/2093-6966-v18-n04-007-g001.jpg

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