PMID- 26204835 OWN - NLM STAT- MEDLINE DCOM- 20160421 LR - 20211203 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 16 IP - 7 DP - 2015 Jul 22 TI - 14-3-3gamma Regulates Lipopolysaccharide-Induced Inflammatory Responses and Lactation in Dairy Cow Mammary Epithelial Cells by Inhibiting NF-kappaB and MAPKs and Up-Regulating mTOR Signaling. PG - 16622-41 LID - 10.3390/ijms160716622 [doi] AB - As a protective factor for lipopolysaccharide (LPS)-induced injury, 14-3-3gamma has been the subject of recent research. Nevertheless, whether 14-3-3gamma can regulate lactation in dairy cow mammary epithelial cells (DCMECs) induced by LPS remains unknown. Here, the anti-inflammatory effect and lactation regulating ability of 14-3-3gamma in LPS-induced DCMECs are investigated for the first time, and the molecular mechanisms responsible for their effects are explored. The results of qRT-PCR showed that 14-3-3gamma overexpression significantly inhibited the mRNA expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-1beta (IL-1beta) and inducible nitric oxide synthase (iNOS). Enzyme-linked immunosorbent assay (ELISA) analysis revealed that 14-3-3gamma overexpression also suppressed the production of TNF-alpha and IL-6 in cell culture supernatants. Meanwhile, CASY-TT Analyser System showed that 14-3-3gamma overexpression clearly increased the viability and proliferation of cells. The results of kit methods and western blot analysis showed that 14-3-3gamma overexpression promoted the secretion of triglycerides and lactose and the synthesis of beta-casein. Furthermore, the expression of genes relevant to nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinase (MAPKs) and lactation-associated proteins were assessed by western blot, and the results suggested that 14-3-3gamma overexpression inactivated the NF-kappaB and MAPK signaling pathways by down-regulating extracellular signal regulated protein kinase (ERK), p38 mitogen-activated protein kinase (p38MAPK) and inhibitor of NF-kappaB (IkappaB) phosphorylation levels, as well as by inhibiting NF-kappaB translocation. Meanwhile, 14-3-3gamma overexpression enhanced the expression levels of beta-casein, mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase 1 (S6K1), serine/threonine protein kinase Akt 1 (AKT1), sterol regulatory element binding protein 1 (SREBP1) and peroxisome proliferator-activated receptor gamma (PPARgamma). These results suggest that 14-3-3gamma was able to attenuate the LPS-induced inflammatory responses and promote proliferation and lactation in LPS-induced DCMECs by inhibiting the activation of the NF-kappaB and MAPK signaling pathways and up-regulating mTOR signaling pathways to protect against LPS-induced injury. FAU - Liu, Lixin AU - Liu L AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. liulixin_2003@163.com. FAU - Lin, Ye AU - Lin Y AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. linlu516@163.com. FAU - Liu, Lili AU - Liu L AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. liulili-liulili@163.com. FAU - Bian, Yanjie AU - Bian Y AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. bianjie.033@163.com. FAU - Zhang, Li AU - Zhang L AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. zhangli_zl18800@163.com. FAU - Gao, Xuejun AU - Gao X AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. gxjgxj112233@163.com. FAU - Li, Qingzhang AU - Li Q AD - Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China. qzli@neau.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150722 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (14-3-3 Proteins) RN - 0 (Caseins) RN - 0 (Interleukins) RN - 0 (Lipopolysaccharides) RN - 0 (NF-kappa B) RN - 0 (Tumor Necrosis Factor-alpha) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases) RN - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases) RN - J2B2A4N98G (Lactose) SB - IM MH - 14-3-3 Proteins/genetics/*metabolism MH - Animals MH - Caseins/genetics/*metabolism MH - Cattle MH - Cells, Cultured MH - Epithelial Cells/*metabolism MH - Extracellular Signal-Regulated MAP Kinases/metabolism MH - Female MH - Humans MH - Interleukins/genetics/metabolism MH - Lactose/metabolism MH - Lipopolysaccharides/pharmacology MH - *MAP Kinase Signaling System MH - Mammary Glands, Animal/cytology/*metabolism MH - NF-kappa B/*metabolism MH - Nitric Oxide Synthase Type II/genetics/metabolism MH - TOR Serine-Threonine Kinases/*metabolism MH - Tumor Necrosis Factor-alpha/genetics/metabolism MH - p38 Mitogen-Activated Protein Kinases/metabolism PMC - PMC4519969 OTO - NOTNLM OT - 14-3-3gamma OT - MAPKs OT - NF-kappaB OT - dairy cow mammary epithelial cells OT - inflammatory responses OT - mTOR signaling pathway EDAT- 2015/07/25 06:00 MHDA- 2016/04/22 06:00 PMCR- 2015/07/01 CRDT- 2015/07/25 06:00 PHST- 2015/04/17 00:00 [received] PHST- 2015/07/05 00:00 [revised] PHST- 2015/07/14 00:00 [accepted] PHST- 2015/07/25 06:00 [entrez] PHST- 2015/07/25 06:00 [pubmed] PHST- 2016/04/22 06:00 [medline] PHST- 2015/07/01 00:00 [pmc-release] AID - ijms160716622 [pii] AID - ijms-16-16622 [pii] AID - 10.3390/ijms160716622 [doi] PST - epublish SO - Int J Mol Sci. 2015 Jul 22;16(7):16622-41. doi: 10.3390/ijms160716622.