克氏原螯虾<i>traf</i>6<i> like</i>基因参与抗细菌先天免疫反应的研究

蔺思函; 杨兵兵; 李倩倩; 沈秀丽; 杜志强

doi:10.19303/j.issn.1008-0384.2022.010.001

克氏原螯虾traf6 like基因参与抗细菌先天免疫反应的研究

Molecular Mechanism of traf6 like Gene in Innate Immunity of Procambarus clarkii

摘要

摘要:
目的深入研究克氏原螯虾（Procambarus clarkia）traf6 like基因参与先天免疫调节的功能。
方法以Pc-traf6 like基因为研究对象，利用RNAi技术干扰目的基因表达量之后，进行细菌模拟刺激，并且通过实时荧光定量PCR（qRT-PCR）检测细胞凋亡相关基因表达量的变化情况。
结果干扰Pc-traf6 like基因，通过干扰效果检测，显示48 h时干扰效果良好，可用于后续试验。在目的基因干扰后，进行金黄色葡萄球菌（Staphylococcus aureus）刺激，检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达；在目的基因干扰后，进行鲶爱德华氏菌（Edwardsiella ictaluri ）刺激，检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达。
结论 Pc-traf6 like基因被干扰，机体在受到金黄色葡萄球菌及鲶爱德华氏菌刺激后，Bax、Bcl 2和Caspase 3基因的相对表达量上调，推测Pc-traf6 like基因通过调控细胞凋亡参与了先天免疫反应。

Abstract:
Objective Role of traf6 like gene in innate immunity regulation of Procambarus clarkia(Pc-traf6 like) was investigated.
Methods RNAi technology was employed to interfere the expression of Pc-traf6 like gene. Bacterial simulation was performed to determine the changes of apoptosis-related gene expression by real-time quantitative PCR (qRT-PCR).
Result The interference on the gene was effective after 48 h for the subsequent experiments. The interference-induced pro-apoptotic gene Bax, anti-apoptotic gene Bcl 2, and apoptotic protease Caspase 3 were upregulated in varying degrees with the stimulation of Staphylococcus aureus or Edwardsiella ictaluri.
Conclusion After the Pc-traf6 like gene was effectively interfered, the relative expressions of Bax, Bcl 2, and Caspase 3 were upregulated after the P. clarkia was stimulated by either S. aureus or E. ictaluri indicating an involvement of Pc-traf6 like gene in the innate immune response by the regulating apoptosis.

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