China Animal Husbandry and Veterinary Medicine ›› 2021, Vol. 48 ›› Issue (5): 1507-1515.doi: 10.16431/j.cnki.1671-7236.2021.05.001

• Biotechnology • Previous Articles     Next Articles

Regulatory Effect of has-miR-5196-3p on Inflammasome During Brucella Infection

LI Jia1, DENG Xingmei1, TAO Tingting1, ZHU Dexin1, QIU Runhui1, WANG Zihang1, WU Jie2, ZHANG Hui1   

  1. 1. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
    2. Animal Husbandry and Veterinary Work Station of the Eighth Division of Xinjiang Production and Construction Corps, Shihezi 832000, China
  • Received:2020-10-15 Online:2021-05-20 Published:2021-05-20

PDF (PC)

81

Abstract: This study was aimed to explore the regulation of the miRNA molecule has-miR-5196-3p on the NOD-like receptor protein 3 inflammasome (NLRP3) during the process of Brucella infecting THP1 cells.Use TargetScan,MiRDB and other bioinformatics prediction sites to predict the binding sites of has-miR-5196-3p and NLRP3-3'UTR.Its wild-type and mutant dual luciferase reporter vectors were construced,respectively,has-miR-5196-3p mimics and negative control were transfected into 293T cells,and the relative luciferase activity changes of each group were detected by the dual luciferase reporter system to verify the targeting relationship between has-miR-5196-3p and NLRP3-3'UTR.THP1 cells were infected with Brucella,the mRNA expressions of NLRP3,caspase1 and has-miR-5196-3p were detected by Real-time PCR,and the protein expressions of NLRP3 and caspase1 were detected by Western blotting..has-miR-5196-3p mimics,has-miR-5196-3p inhibitor and negative control were transfected into THP1 cells,Real-time PCR and Western blotting were used to detect the expression level of NLRP3 and cysteinyl aspartate specific protease (caspase1).The results showed that the wild-type and mutant dual luciferase reporter vector containing NLRP3-3'UTR was successfully constructed.has-miR-5196-3p mimics extremely significantly reduced the relative luciferase activity of pmirGLO-NLRP3-3'UTR-wt(P<0.01).After Brucella infecting THP1 cells,NLRP3 inflammasome was activated,while has-miR-5196-3p did not change significantly (P>0.05).has-mi-R-5196-3p mimics could significantly inhibit the expression of NLRP3 (P<0.05),while has-miR-5196-3p inhibitor was the opposite.It showed that the miRNA molecule has-miR-5196-3p could bind to NLRP3-3'UTR and inhibit its expression.During the process of Brucella infecting THP1 cells,has-miR-5196-3p targets and negatively regulated NLRP3 inflammasome expression and inflammation happened.

Key words: Brucella; regulation; has-miR-5196-3p; NLRP3

CLC Number: