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首页> 外文期刊>British Biotechnology Journal >Apoptotic Gene Expression in Sheep Hepatocytes during Fasciola hepatica Infection (Fascioliasis)
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Apoptotic Gene Expression in Sheep Hepatocytes during Fasciola hepatica Infection (Fascioliasis)

机译：肝片Fasciola感染过程中绵羊肝细胞中凋亡基因的表达（Fascioliasis）

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Aims: The main objective of this work was to investigate the apoptotic genes of sheep liver hepatocytes to elucidate the apoptosis pathway mechanisms during Fasciola hepatica infection using molecular and serological techniques. Study Design: This is a laboratory based study whereby F. hepatica infected liver specimens were used. Methodology: Total RNA was extracted from fresh-frozen liver tissue. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to investigate the genes encoding the mRNA for following proteins: 18S; Bax; Bcl-2; Caspase-3. Additionally, the concentration of total protein in each sample was estimated spectrophotometrically. Polyclonal antibodies were produced by immunizing rabbits and diluted with 5% skimmed milk in PBS containing 0.1% Tween 20 (1:1000). The detection of the reacted antigen/antibody products was performed using immunoblotting technique were used to assess the quantification of protein kinetic to apoptotic genes. Data obtained from this investigation were analyzed using Portable IBM SPSS 2006 Statistics software. Results: We investigated the apoptotic gene expression in sheep liver hypatocytes during F. hepatica infection by mRNA expression of three genes involved in apoptosis; Bax, Bcl-2 and Caspase-3 and histopathological parameter after infection with F. hepatica in the liver cells. Quantitative real-time PCR, histological examination and immunoblotting were used to quantify apoptotic genes, histopathology and protein kinetic, respectively. F. hepatica infection induces apoptosis in the liver cells via Bax, Bcl-2 and Caspase-3 tests. Conclusions: F. hepatica infection induces apoptosis in the liver cells via Bax, Bcl-2 and Caspase-3 tests and it precedes necrosis. Thus, this study suggests that the induced apoptotic gene expression was due to the outcome of F. hepatica.
机译：目的：这项工作的主要目的是使用分子和血清学技术研究绵羊肝肝细胞的凋亡基因，以阐明在肝片状Fasciola感染过程中的凋亡途径机制。研究设计：这是一项基于实验室的研究，其中使用了感染肝炎链球菌的肝标本。方法：从新鲜冷冻的肝组织中提取总RNA。进行实时定量聚合酶链反应（qRT-PCR）以研究编码以下蛋白的mRNA的基因：18S;巴克斯Bcl-2; Caspase-3。另外，通过分光光度法估算每个样品中总蛋白质的浓度。通过免疫兔子产生多克隆抗体，并在含0.1％Tween 20（1：1000）的PBS中用5％脱脂牛奶稀释。使用免疫印迹技术对反应的抗原/抗体产物进行检测，以评估对凋亡基因的蛋白质动力学定量。使用Portable IBM SPSS 2006 Statistics软件分析了从该调查中获得的数据。结果：我们通过三个参与凋亡的基因的mRNA表达研究了肝炎链球菌感染过程中绵羊肝肝细胞凋亡基因的表达。肝细胞感染肝炎双歧杆菌后Bax，Bcl-2和Caspase-3及其组织病理学参数实时定量PCR，组织学检查和免疫印迹法分别用于量化凋亡基因，组织病理学和蛋白质动力学。肝炎性肝炎感染通过Bax，Bcl-2和Caspase-3检测诱导肝细胞凋亡。结论：肝炎性肝炎感染通过Bax，Bcl-2和Caspase-3检测诱导肝细胞凋亡，并在坏死之前发生。因此，该研究表明诱导的凋亡基因表达是由于肝小球藻的结果。

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