GasderminA3基因在小鼠皮肤上皮中对NF-κB表达的影响  

Gasdermin A3 affects NF-κB expression in mouse skin keratinocytes

作　　者：余裕[1] 白秀峰[1] 赖向东[1] 杨恬[1] 连小华[1] 

机构地区：[1]第三军医大学基础医学部细胞生物学教研室,重庆400038

出　　处：《第三军医大学学报》2014年第4期313-316,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81171515)~~

摘　　要：目的探讨Gasdermin A3(简写为Gsdma3)基因在小鼠皮肤上皮中对NF-κB表达的影响。方法通过组织切片与免疫荧光观察Gasdermin A3突变鼠与正常鼠背部皮肤中NF-κB蛋白的阳性表达区域与强弱情况;免疫荧光观察转染Gsdma3质粒后,小鼠皮肤细胞中Gsdma3与NF-κB蛋白的表达情况;RT-PCR检测Gsdma3基因突变或过表达的情况下,在体或离体小鼠皮肤上皮细胞中NF-κB mRNA的表达变化情况;Western blot检测Gsdma3基因突变或过表达的情况下,在体或离体小鼠皮肤上皮细胞中NF-κB蛋白的表达变化情况。结果在小鼠皮肤上皮细胞中,Gsdma3基因突变导致小鼠皮肤上皮细胞中NF-κB mRNA表达平均降低39.53%,(P<0.01),蛋白表达平均降低65.03%(P<0.01),免疫荧光检测NF-κB阳性表达在Gsdma3突变的皮肤上皮中明显减弱;Gsdma3的过表达导致小鼠皮肤上皮细胞中NF-κB mRNA表达平均增强56.25%(P<0.01),蛋白表达平均增强199.6%(P<0.01),免疫荧光检测NF-κB阳性表达在过表达Gsdma3的皮肤上皮细胞中明显增强。结论在小鼠皮肤上皮中,Gsdma3能促进NF-κB的表达。Objective To determine the effect of gasdermin A3 （Gsdma3） gene on NF-κB expression in mouse skin keratinocytes.Methods Histological section and immunofluorescent assays were performed to locate and measure NF-κB expression between the Gsdma3-mutant mouse skin and the normal one. NF-κB expression was revealed by immunofluorescent assay after Gsdma3 expression plasmid was transfected into mouse skin keratinocytes. RT-PCR and Western blotting were used in vivo and in vitro to investigate NF-κB expression at mRNA and protein levels, respectively, in Gsdma3-mutant or Gsdma3-overexpressed skin keratinocytes.Results In mouse skin keratinocytes, Gsdma3 mutation decreased NF-κB mRNA expression by 39.53% （P〈0.01） and protein expression by 65.03% （P〈0.01）. Positive zones and intensity of immunofluorescence were significantly mitigated in Gsdma3-mutant skin keratinocytes as compared to the normal ones. On the other hand, overexpression of Gsdma3 could increase the mRNA expression of NF-κB by 56.25% （P〈0.01） and the protein expression by 199.6% （P〈0.01）. In addition, Gsdma3 and NF-κB exhibited an opposite expression manner in the in vitro immunofluorescent assays.Conclusion Gsdma3 can induce mRNA and protein expression of NF-κB in the mouse skin keratinocytes.

关 键 词：Gasdermin A3 NF-ΚB 小鼠 皮肤上皮细胞 

分 类 号：R322.99[医药卫生—人体解剖和组织胚胎学] R394-33[医药卫生—基础医学]