Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery · 2015
Expression of Surfactant Protein-A during LPS-Induced Otitis Media with Effusion in Mice.
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Li Li, Guo Xiangrui, Olszewski Emily, Fan Zhaomin, Ai Yu, Han Yuechen, Xu Lei, Li Jianfeng, Wang Haibo
Аннотация
OBJECTIVE: The objective of this study was to investigate the expression and role of surfactant protein (SP) in the middle ear throughout lipopolysaccharide (LPS)-induced otitis media with effusion (OME).
STUDY DESIGN: Randomized case-controlled animal model.
SETTING: Shandong University, Jinan, China.
SUBJECTS AND METHODS: SP expression was monitored using reverse transcription polymerase chain reaction (PCR) in normal mice (n = 5). Two groups, control phosphate-buffered saline-injected mice (n = 5) and LPS-injected mice (n = 5), were euthanized 5 days following injection. RNA was extracted for reverse transcription PCR and real-time PCR, and temporal bone samples were used for hematoxylin and eosin staining. LPS was injected into mice, and 5 mice per test were euthanized at 0, 12, 24, 48, 72, and 96 hours following injection. For mRNA expression quantification, reverse transcription PCR and real-time PCR were performed, and proinflammatory cytokine levels were measured by enzyme-linked immunosorbent assay.
RESULTS: SP-A and SP-D expression was detected in normal murine Eustachian tubes. SP-A expression was up-regulated after LPS-induced OME (P = .01). At various time points after LPS injection, concentrations of proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin (IL)-1β, and IL-6) in the middle ear increased significantly (P < .05) and correlated with changes in SP-A expression.
CONCLUSION: SP-A and SP-D exist in the murine middle ear. The expression of SP-A and TNF-α, IL-1β, and IL-6 was up-regulated in the middle ear of the LPS-induced OME animal model.
STUDY DESIGN: Randomized case-controlled animal model.
SETTING: Shandong University, Jinan, China.
SUBJECTS AND METHODS: SP expression was monitored using reverse transcription polymerase chain reaction (PCR) in normal mice (n = 5). Two groups, control phosphate-buffered saline-injected mice (n = 5) and LPS-injected mice (n = 5), were euthanized 5 days following injection. RNA was extracted for reverse transcription PCR and real-time PCR, and temporal bone samples were used for hematoxylin and eosin staining. LPS was injected into mice, and 5 mice per test were euthanized at 0, 12, 24, 48, 72, and 96 hours following injection. For mRNA expression quantification, reverse transcription PCR and real-time PCR were performed, and proinflammatory cytokine levels were measured by enzyme-linked immunosorbent assay.
RESULTS: SP-A and SP-D expression was detected in normal murine Eustachian tubes. SP-A expression was up-regulated after LPS-induced OME (P = .01). At various time points after LPS injection, concentrations of proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin (IL)-1β, and IL-6) in the middle ear increased significantly (P < .05) and correlated with changes in SP-A expression.
CONCLUSION: SP-A and SP-D exist in the murine middle ear. The expression of SP-A and TNF-α, IL-1β, and IL-6 was up-regulated in the middle ear of the LPS-induced OME animal model.