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在幼年和成年蛇肝脏中具有潜在抗蛇毒作用的抑制剂的差异转录谱。

Differential transcript profile of inhibitors with potential anti-venom role in the liver of juvenile and adult snake.

作者信息

Gomes Cícera Maria, de Morais-Zani Karen, Lu Stephen, Buarque Diego de Souza, Braz Glória Regina Cardoso, Grego Kathleen Fernandes, Tanaka Aparecida Sadae, Tanaka-Azevedo Anita Mitico

机构信息

Laboratório de Herpetologia, Instituto Butantan, São Paulo, Brazil.

Interunidades em Biotecnologia, Universidade de São Paulo, São Paulo, Brazil.

出版信息

PeerJ. 2017 Apr 27;5:e3203. doi: 10.7717/peerj.3203. eCollection 2017.

Abstract

BACKGROUND

Snakes belonging to the genus are vastly distributed in Central and South America and are responsible for most cases of reported snake bites in Latin America. The clinical manifestations of the envenomation caused by this genus are due to three major activities-proteolytic, hemorrhagic and coagulant-mediated by metalloproteinases, serine proteinases, phospholipases A and other toxic compounds present in snake venom. Interestingly, it was observed that snakes are resistant to the toxic effects of its own and other snake's venoms. This natural immunity may occur due the absence of toxin target or the presence of molecules in the snake plasma able to neutralize such toxins.

METHODS

In order to identify anti-venom molecules, we construct a cDNA library from the liver of snakes. Moreover, we analyzed the expression profile of four molecules-the already known anti-hemorrhagic factor Bj46a, one gamma-phospholipase A inhibitor, one inter-alpha inhibitor and one C1 plasma protease inhibitor-in the liver of juvenile and adult snakes by qPCR.

RESULTS

The results revealed a 30-fold increase of gamma-phospholipase A inhibitor and a minor increase of the inter-alpha inhibitor (5-fold) and of the C1 inhibitor (3-fold) in adults. However, the Bj46a factor seems to be equally transcribed in adults and juveniles.

DISCUSSION

The results suggest the up-regulation of different inhibitors observed in the adult snakes might be a physiological adaptation to the recurrent contact with their own and even other snake's venoms throughout its lifespan. This is the first comparative analysis of ontogenetic variation of expression profiles of plasmatic proteins with potential anti-venom activities of the venomous snake . Furthermore, the present data contributes to the understanding of the natural resistance described in these snakes.

摘要

背景

该属蛇类广泛分布于中美洲和南美洲,是拉丁美洲大多数报告的蛇咬伤病例的元凶。该属蛇毒所致中毒的临床表现归因于三种主要活性——由金属蛋白酶、丝氨酸蛋白酶、磷脂酶A及蛇毒中存在的其他有毒化合物介导的蛋白水解、出血和凝血活性。有趣的是,人们观察到蛇对自身及其他蛇类毒液的毒性作用具有抗性。这种天然免疫力可能是由于缺乏毒素靶点,或者蛇血浆中存在能够中和此类毒素的分子。

方法

为了鉴定抗毒分子,我们构建了该属蛇肝脏的cDNA文库。此外,我们通过定量PCR分析了四种分子(已知的抗出血因子Bj46a、一种γ-磷脂酶A抑制剂、一种α-抑制剂和一种C1血浆蛋白酶抑制剂)在幼年和成年蛇肝脏中的表达谱。

结果

结果显示,成年蛇体内γ-磷脂酶A抑制剂增加了30倍,α-抑制剂(增加了5倍)和C1抑制剂(增加了3倍)略有增加。然而,Bj46a因子在成年蛇和幼年蛇中的转录情况似乎相同。

讨论

结果表明,成年蛇体内观察到的不同抑制剂上调可能是一种生理适应,以应对其一生中反复接触自身甚至其他蛇类的毒液。这是首次对具有潜在抗蛇毒活性的血浆蛋白表达谱的个体发育变异进行比较分析。此外,目前的数据有助于理解这些蛇类所具有的天然抗性。

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