siRNA抑制牛子宫内膜上皮细胞Nrf2基因的表达研究

doi:10.16431/j.cnki.1671-7236.2019.06.001

Abstract

Abstract:

In order to investigate the effect of nuclear factor E2 related factor (Nrf2) gene on bovine endometrial epithelial cells,this experiment used small interference RNA (siRNA) technology and tert-butylhydroquinone(tBHQ) as Nrf2 activator to study the effects of Nrf2 on the expression of hemeoxygenase-1 (HO-1) and Homebox A10 (HOXA10) in bovine endometrial epithelial cells.The results showed that in the designed two siRNA sequences(siRNA-1209 and siRNA-1672),the siRNA-1672 had a better inhibitory effect at a final concentration of 75 nmol/L and a response time of 24 h by Real-time quantitative PCR,the inhibition efficiency was over 80%.Western blotting results showed that the protein expression level of Nrf2 extremely significantly decreased at 96 h after transfection (P<0.01);While the mRNA expression levels of HO-1 and HOXA10 extremely significantly decreased by 60% and 70% (P<0.01),respectively,and the protein expression of HO-1 and HOXA10 extremely significantly or significantly decreased after 96 h (P<0.01;P<0.05).In addition,CCK8 method was used in this test,it found that the proliferation ability of bovine endometrial epithelial cells was weakened.In the tBHQ-activated the expression of Nrf2 assay,the protein expression of Nrf2 was the highest in the final concentration of tBHQ at 30 μmol/L,which was extremely significantly higher than that of control group (P<0.01),and the protein expression levels of HO-1 and HOXA10 were also higher than that of control group.This experiment results indicated that the designed siRNA-1672 could specifically inhibit the expression of Nrf2 in bovine endometrial epithelial cells,while the inhibition of Nrf2 expression led to a decrease in the proliferation ability of bovine endometrial epithelial cells,and Nrf2 regulated the expression of HO-1 and HOXA10 genes in bovine endometrial epithelial cells.

Key words: bovine; RNAi; endometrial epithelial cell; Nrf2 gene; HO-1 gene; HOXA10 gene

CLC Number:

S814.8

HU Xiuzhong, CHENG Lei, YU Jie, XIANG Min, XIA Yu, TAO Bifei, ZHOU Yuan, WANG Dingfa, XIONG Haiqian. Study on Inhibition of Nrf2 Gene Expression in Bovine Endometrial Epithelial Cells by siRNA[J]. , 2019, 46(6): 1567-1575.

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Study on Inhibition of Nrf2 Gene Expression in Bovine Endometrial Epithelial Cells by siRNA

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