Xingmei Deng1, Jia Guo1, Zhihua Sun1, Laizhen Liu2, Tianyi Zhao1, Jia Li1, Guochao Tang3, Hai Zhang4, Wenjing Wang4, Shuzhu Cao1, Dexin Zhu1, Tingting Tao1, Gang Cao5, P I Baryshnikov6, Chuangfu Chen1, Zongsheng Zhao1, Lihua Chen7, Hui Zhang1. 1. State International Joint Research Center for Animal Health Breeding, College of Animal Science and Technology, Shihezi University, Shihezi, China. 2. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi, China. 3. Technology Center, Xinjiang Tianrun Dairy Biological Products Co., Ltd, Urumqi, China. 4. Department of Transfusion Medicine, Southern Medical University, Guangzhou, China. 5. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China. 6. College of Veterinary, Altai National Agricultural University, Barnaul, Russia. 7. College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, China.
Abstract
Objectives: The underlying mechanism of the inflammatory response against Brucellosis caused by Brucella remains poorly understood. This study aimed to determine the role of long non-coding RNAs (lncRNAs) in regulating of inflammatory and anti-Brucella responses. Materials and methods: Microarray analysis was performed to detect differentially expressed lncRNAs in THP-1 cells infected with an S2308 Brucella strain. The candidate lncRNAs were screened using bioinformatic analysis and siRNAs; bioinformatic prediction and luciferase reporter assay were also conducted, while inflammatory responses was assessed using RT-qPCR, western blot, immunofluorescence, ELISA, HE, and immunohistochemistry. Results: The lncRNA Gm28309 was identified to be involved in regulating inflammation induced by Brucella. Gm28309, localized in the cytoplasm, was down-expressed in RAW264.7 cells infected with S2308. Overexpression of Gm28309 or inhibition of miR-3068-5p repressed p65 phosphorylation and reduced NLRP3 inflammasome and IL-1β and IL-18 secretion. Mechanistically, Gm28309 acted as a ceRNA of miR-3068-5p to activate NF-κB pathway by targeting κB-Ras2, an inhibitor of NF-κB signaling. Moreover, the number of intracellular Brucella was higher when Gm28309 was overexpressed or when miR-3068-5p or p65 was inhibited. However, these effects were reversed by the miR-3068-5p mimic. Conclusions: Our study demonstrates, for the first time, that LncRNAs are involved in regulating immune responses during Brucella infection, and Gm28309, an lncRNA, plays a crucial role in activating NF-κB/NLRP3 inflammasome signaling pathway.
Objectives: The underlying mechanism of the inflammatory response against Brucellosis caused by Brucella remains poorly understood. This study aimed to determine the role of long non-coding RNAs (lncRNAs) in regulating of inflammatory and anti-Brucella responses. Materials and methods: Microarray analysis was performed to detect differentially expressed lncRNAs in THP-1 cells infected with an S2308 Brucella strain. The candidate lncRNAs were screened using bioinformatic analysis and siRNAs; bioinformatic prediction and luciferase reporter assay were also conducted, while inflammatory responses was assessed using RT-qPCR, western blot, immunofluorescence, ELISA, HE, and immunohistochemistry. Results: The lncRNA Gm28309 was identified to be involved in regulating inflammation induced by Brucella. Gm28309, localized in the cytoplasm, was down-expressed in RAW264.7 cells infected with S2308. Overexpression of Gm28309 or inhibition of miR-3068-5p repressed p65 phosphorylation and reduced NLRP3 inflammasome and IL-1β and IL-18 secretion. Mechanistically, Gm28309 acted as a ceRNA of miR-3068-5p to activate NF-κB pathway by targeting κB-Ras2, an inhibitor of NF-κB signaling. Moreover, the number of intracellular Brucella was higher when Gm28309 was overexpressed or when miR-3068-5p or p65 was inhibited. However, these effects were reversed by the miR-3068-5p mimic. Conclusions: Our study demonstrates, for the first time, that LncRNAs are involved in regulating immune responses during Brucella infection, and Gm28309, an lncRNA, plays a crucial role in activating NF-κB/NLRP3 inflammasome signaling pathway.
Authors: C Freudlsperger; Y Bian; S Contag Wise; J Burnett; J Coupar; X Yang; Z Chen; C Van Waes Journal: Oncogene Date: 2012-05-28 Impact factor: 9.867
Authors: Ya Pan; Wenjuan Chen; Xiangyun Yan; Boshi Yu; Shuwen Yao; Xiaohui Chen; Shuping Han Journal: Biomed Res Int Date: 2021-11-03 Impact factor: 3.411