细粒棘球绦虫BAG3和EB1基因的克隆、表达及其在原头蚴细胞凋亡中的作用

doi:10.11843/j.issn.0366-6964.2022.05.024

Abstract

Abstract: To explore the molecular characteristics of Echinococcus granulosus BAG3 (Eg-BAG3) and EB1 (Eg-EB1) and their roles in the apoptosis of protoscoleces, recombinant Eg-BAG3 and Eg-EB1were expressed in E. coli BL21(DE3), and bioinformatic analysis, Western blotting and immunofluorescence localization were conducted to explore the molecular characteristics of Eg-BAG3 and Eg-EB1. The apoptosis of protoscoleces was induced by 10 mmol·L^-1H₂O₂ and the transcription levels of Eg-BAG3 and Eg-EB1 were detected by qRT-PCR, and their roles in the apoptosis of protoscoleces was further verified by RNA interference (RNAi). Eg-BAG3 contained the characteristic BAG domain of BAG protein family and Eg-EB1 contained the characteristic BAR domain of endophilin protein family, suggesting that Eg-BAG3 and Eg-EB1 were the typical BAG protein and endophilin protein, respectively. Immunolocalization showed that Eg-BAG3 was widely distributed in all development stages of E. granulosus,Eg-EB1 was mainly located in the tegument, suckers and rostellum of protoscoleces and germinal layer, while no positive signal was observed in adults. Protoscoleces apoptosis was successfully induced by 10 mmol·L^-1H₂O₂ and the relative transcription levels of Eg-BAG3 and Eg-EB1 significantly increased in response to H₂O₂ treatment. RNA interference (RNAi) analysis showed that the apoptosis rate of protoscoleces in the Eg-BAG3 interference group was significantly increased compared with non-interference group (P<0.05), while the Eg-EB1 interference group showed a lower apoptosis rate of protoscoleces than non-interference group (P<0.05), suggesting that Eg-BAG3 inhibited protoscoleces apoptosis induced by H₂O₂, while Eg-EB1 promoted protoscoleces apoptosis. In conclusion, Eg-BAG3 could inhibit protoscoleces apoptosis induced by oxidative stress, while Eg-EB1 promotes apoptosis, and both Eg-BAG3 and Eg-EB1 are possibly involved in regulating the formation of infertile cysts.

Key words: Echinococcus granulosus, BAG3, EB1, apoptosis, RNA interference

CLC Number:

S852.734

HE Xue, WANG Ning, HUA Ruiqi, DU Xiaodi, YANG Guangyou. Cloning, Expression of Echinococcus granulosus BAG3 and EB1 and Their Roles in the Apoptosis of Protoscoleces[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1562-1575.

References 32

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Cloning, Expression of Echinococcus granulosus BAG3 and EB1 and Their Roles in the Apoptosis of Protoscoleces

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