RESUMO
Mammary epithelial cells (MECs) play an important role in debating Streptococcus uberis (S. uberis) infection. Toll like receptor (TLR) engagement leads to the recruitment of phosphatidylinositol 3 kinases (PI3K). In order to investigate the relationship of TLRs/NF-κB and PI3K/Akt/mTOR signaling pathways in S. uberis infection in MECs, we challenged MECs (EpH4-Ev) with S. uberis 0140 J and quantified the adaptor molecules in these two signaling pathways, as-well-as proinflammatory cytokines and cell damage. The results indicate that the host's responses to virulent S. uberis infection are complex. In MECs, both TLR2 and TLR4 are detecting S. uberis infection and TLR2 is the principal receptor. The role of the PI3K/Akt/mTOR pathway in inflammatory regulation is independent of the activation of TLRs/NF-κB. Cross-talk between PI3K/Akt/mTOR and TLRs/NF-κB signaling pathways promote inflammation. This study increases our understanding of the molecular defense mechanisms of MECs in S. uberis mastitis, and provides theoretical support for the prevention of this disease.
Assuntos
Glândulas Mamárias Animais/microbiologia , NF-kappa B/imunologia , Fosfatidilinositol 3-Quinases/imunologia , Proteínas Proto-Oncogênicas c-akt/imunologia , Streptococcus/imunologia , Serina-Treonina Quinases TOR/imunologia , Receptores Toll-Like/imunologia , Animais , Bovinos , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Feminino , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Inflamação/microbiologia , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/imunologia , Mastite/imunologia , Mastite/microbiologia , NF-kappa B/genética , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/imunologia , Streptococcus/patogenicidade , Serina-Treonina Quinases TOR/genética , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Receptores Toll-Like/genéticaRESUMO
OBJECTIVE: To investigate the effect of genistein on cell proliferation and apoptosis in human laryngeal carcinoma cell line Hep-2. METHOD: Cell Counting Kit-8 (CCK-8) assay was used to measure the 50% inhibiting concentration (IC50) value of genistein; cell apoptosis rate and the distribution changes of cell cycle were determined with flow cytometry assay after treatment by gensitein. The morphological changes of tumor cells were evaluated by inverted phase contrast mircroscopy. RESULT: The IC50 of geniste responses to Hep-2 cells for 24 h was 23.64 µg/ml. The apoptotic rates of Hep-2 cells treated by genistein for 24 h were 22.40% ± 1. 65% (at 12 µg/ml genistein) and 30.64% ± 2.94% (at 24 µg/ml genistein) respectively, significantly statistical differences were foundbetween above threated groups and the control group (P < 0.05); the apoptotic rates of Hep-2 cells treated by genistein for 48 h were 30.55% ± 0.72%(at 12 µg/ml genistein) and 48.69% ± 1.06% (at 24 µg/ml genistein) respectively, significantly statistical differences were found between above threated groups and the control group (P < 0.05). When Hep-2 cells exposed to the same concentration of genistein for 24 h, 48 h respectively, the difference in apoptotic rate was statistically significant. CONCLUSION: Genistein inhibited Hep-2 cells growth obviously, meanwhile it could induced apoptosis of Hep-2 cells, the apoptotic rate was increasing with the increase of the time and dose of genistein.