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The Anti-inflammatory Mechanism and the Effect on the Mucin5ac Production of ELAFIN in Cytoplasm

Author: WeiHuaSheng
Tutor: ZhouXiangDong
School: Chongqing Medical University
Course: Internal Medicine
Keywords: Elafin nucleat factor-κb small ubiquitin-like modifier -1 anti-inflammatory action mucin5AC
CLC: R563.9
Type: Master's thesis
Year: 2011
Downloads: 14
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Abstract


Objective: To investigate the mechanism of Elafin anti-infla- mmatory action and the effect of it on mucin(MUC)5AC production in cytoplasm.Methods: (1) After cultivating 16HBE cells in vitro, the constructed pEGFP -N1-Elafin eukaryotic expression vector was transfected into the cells, and empty plasmid as a vector control. The cells were then incubated with exogenous TNF-α.The cells were divided into 3 groups:①b lank control group;②empty plasmid+TNF-α(10μg/L) group ;③pEGFP-N1-Elafin+ TNF-α(10μg/L) group. The NF-κB activity in nuclear was detected by immunofluorescence and laser scanning confocal microscope,the IL-1βand IL-8 production was assayed by ELISA, the SUMO-1-p-IκBαproduction was assayed by Western blot. (2) The cells were incubated with exogenous EGF.The cells were divided into 3 groups:①blank control group;②empty plasmid+EGF(25μg/L) group ;③pEGFP-N1-Elafin+ EGF(25μg/L) group. The mucin(MUC)5AC mRNA level was analyzed by RT-PCR,the MUC5AC protein production was assayed by ELISA,the NF-κB activity in nuclear was detected by immunofluorescence and laser scanning confocal microscope,the SUMO-1-p-IκBαproduction was assayed by Western blot.Result: (1) The NF-κB activity,the IL-1βand IL-8 production of the experimental group were significantly lower than those of the empty plasmid group,the SUMO-1-p-IκBαproduction of the experimental group was significantly higher than that of the empty plasmid group(all P﹤0.01). (2) The MUC5AC mRNA level,the MUC5AC protein production and the NF-κB activity of the experimental group were all significantly lower than those of the empty vector group,but the SUMO-1-p-IκBαproduction was significantly higher than that of the latter(all P﹤0.01).Conclusion:(1)Elafin prevents NF–κB activation via an effect on IκBαsumoylation,and it then reduce proinflammatory cytokine production,thus playing a role of anti-inflammatory. (2) Elafin prevents NF–κB activation via an effect on p-IκBαsumoylation,thus reducing epidermal growth factor-induced mucin5AC production.

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CLC: > Medicine, health > Internal Medicine > Respiratory system and chest diseases > Pulmonary disease > Other
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