含CpG-ODN基序重组抑制素质粒的构建及其对小鼠颗粒细胞相关凋亡基因表达的影响

doi:10.11843/j.issn.0366-6964.2018.08.007

Abstract

Abstract:

The purpose of this study was to construct a recombinant inhibin plasmid containing a CpG-ODN fragment with immunogenicity(named as pEGISI-CpG-ODN), and determine its effect on mRNA expression levels of apoptosis-related genes to investigate the roles of CpG-ODN in mouse ovarian granulosa cells (GCs). GCs were obtained from the ovaries of 21-23 days female ICR mice. The experiment was divided into 3 groups:empty vector group (pEGFP), inhibin recombinant plasmid group (pEGISI) and containing CpG-ODN motif recombinant inhibin plasmid group (pEGISI-CpG-ODN). The target fragment CpG-ODN was amplified by PCR. The mRNA expression levels of CpG and the death receptor pathway related genes (Fas, FasL, DR4/5, TRAIL, Caspase8 and Caspase3) in transfected cells were analyzed by quantitative real-time PCR (qRT-PCR). The results showed that a new recombinant inhibin plasmid called pEGISI-CpG-ODN was constructed successfully. Moreover, CpG gene could be highly expressed in GCs after transfection of GCs with recombinant inhibin plasmid (pEGISI-CpG-ODN). Furthermore, the mRNA expression levels of pro-apoptotic genes Fas, FasL, DR4/5,TRAIL, Caspase8 and Caspase3 were very significantly lower in GCs of the pEGISI-CpG-ODN plasmid treated group than those in pEGISI treated group (P<0.01). These results suggested that CpG-ODN might not only resist the function of inhibin but also decrease the expression levels of apoptotic genes in mouse GCs, as a result, it was able to affect follicular development.

CLC Number:

S854.4

YANG Mei-xia, ZHANG Hong-liang, LI Lan-qi, ZHANG Yu-fang, YANG Li-guo, WANG Shui-lian. Construction of Recombinant Inhibin Plasmid Containing CpG-ODN Motif and Its Effect on the Expressions of Apoptosis-related Genes in Mouse Granulosa Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1625-1632.

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Construction of Recombinant Inhibin Plasmid Containing CpG-ODN Motif and Its Effect on the Expressions of Apoptosis-related Genes in Mouse Granulosa Cells

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