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The Relative Study of the Expression of COX-2 and 5-LOX in Respiratory Syncytial Virus-Infected Macrophages

Author: LuMin
Tutor: HuangShengHai
School: Anhui Medical University,
Course: Pathogen Biology
Keywords: Respiratory syncytial virus Cyclooxygenase-2 5 - lipoxygenase Nuclear factor kappa B Macrophages
CLC: R373
Type: Master's thesis
Year: 2009
Downloads: 39
Quote: 0
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Abstract


Objective: To investigate the respiratory syncytial virus (respiratory syncytial virus, RSV) infection lung macrophage RAW264.7 cells, cyclooxygenase -2 (the spectrum of effects of [cyclooxygenase-2, COX-2), 5 - lipoxygenase enzyme (5 - lipoxygenase, 5-LOX) expression of inflammatory mediators levels initially clarify the mechanism of induced expression regulation mechanism for the study of RSV infection inflammation, clinical treatment of RSV infection caused by the inflammatory response to provide a theoretical basis. Method: ① RAW264.7 macrophages RSV infection in vitro, respectively, within the type of infection time point (4h, 8h, 16h, 24h) to collect cells and cell culture supernatant. By semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western-blot were used to detect ethylene dismutase -2 (COX-2) and 5 - lipoxygenase (5-LOX) mRNA and protein expression in the cells, The enzyme-linked immunosorbent assay (ELISA) to detect the cell culture supernatant PGE2 and LTB4 content. ② RSV infection was added 50μmol / L NF-κB specific inhibitor dithiophosphate amino-pyrrolidin-(pyrrolidine dithiocarbamate PDTC) into the nuclear activation, inhibition of NF-kappa B is detected using the same method as above-described cell COX-2 and 5-LOX expression and cell culture supernatant of PGE2 and LTB4 changes. Results: (1) RSV infection after 8h raised RAW264.7 macrophages COX-2 mRNA transcript and protein levels of expression. Cell culture supernatant of PGE2 content increased. Changes in each index were significant differences compared with normal controls, and with the increased expression of RSV infection in the extension of time. The ② RSV infection within 24 hours, the 5-LOX expression RSW264.7 macrophages and cell culture supernatant content of LTB4 than normal control group had no significant difference. (3) RSV infection of PDTC can inhibit NF-kappa B nuclear activation of nuclear NF-kappa B protein expression was significantly reduced; expression of COX-2 transcription and protein levels were significantly lower, that is significantly lower infection 8h, reduce significant difference. 5-LOX transcriptional level and protein level than the control group had no significant difference, reduce downstream inflammatory factors LTB4 content. Conclusion: RSV infected RAW264.7 macrophages early to induce the expression of COX-2, at the same time generate a lot of PGE2 synthesis of PGE2 is mainly affected by the regulation of COX-2, NF-kappaB activation for COX-2 gene expression role in regulating; RSV infection RAW264.7 macrophages within 24 hours of 5-LOX expression had no effect, while the NF-kappa B inhibitors also had no effect on 5-LOX expression and inflammatory factors.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Human Virology ( pathogenic virus)
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