干扰和过表达BAMBI基因对猪卵泡颗粒细胞表型和功能的影响

doi:10.16431/j.cnki.1671-7236.2021.02.018

Abstract

Abstract: To investigate the regulation of bonemorphogenetic protein and activin membrane-bound inhibitor (BAMBI) gene in transforming growth factor-β (TGF-β) signaling pathway on porcine follicular granulosa cells,in this study,BAMBI interference vector and overexpression vector were designed and constructed.Porcine ovarian granulosa cells were ransfected with the constructed interference (pSIREN-BAMBI-sh1,pSIREN-BAMBI-sh2,pSIREN-BAMBI-sh3) and overexpression (pcDNA3.1-BAMBI) recombinant plasmids,and Real-time quantitative PCR technology was used to screen effective fragments and detect mRNA expression levels of TGF-β signaling pathway downstream genes (TGF-βRⅠ,TGF-βRⅡ,SMAD1,SMAD2,SMAD3,SMAD4,SMAD5) and apoptosis genes (Bax,Bcl-2).Finally,the effects of BAMBI interference and overexpression on the proliferation and apoptosis of ovarian granulosa cells were detected by MTT method and flow cytometry.The results showed that pSIREN-BAMBI-sh2 had the best effect of inhibiting BAMBI expression and the highest interference efficiency.When interfering with BAMBI,the expression of TGF-βRⅡ increased significantly (P<0.05),so that the expression of SMAD2 and SMAD3 increased significantly (P<0.05).When overexpressing of BAMBI,TGF-βRⅡ decreased significantly (P<0.05),so that the expression of SMAD2 and SMAD3 decreased significantly (P<0.05).Up-regulation of BAMBI inhibited the proliferation of granulosa cells and significantly promoted apoptosis (P<0.01).The results suggested that the BAMBI gene significantly affected the phenotype and function of ovarian granulosa cells,and might indirectly affect the reproductive performance of pigs by regulating the TGF-β signaling pathway.

Key words: pig; transforming growth factor-β (TGF-β); BAMBI gene; granulosa cells; interference; overexpression

CLC Number:

Q782

SUN Yishan, YANG Fei, HUANG Tao. Effects of Interference and Overexpression of BAMBI Gene on the Phenotype and Function of Porcine Ovarian Granulosa Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 563-573.

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Effects of Interference and Overexpression of BAMBI Gene on the Phenotype and Function of Porcine Ovarian Granulosa Cells

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