Gsdermin A3基因在小鼠皮肤角质形成细胞中对Krt6b基因表达的影响

Gsdermin A3 suppresses Krt6b expression in mouse skin keratinocytes

目的初步探讨Gsdermin A3(Gsdma3)基因对Krt6b基因表达的影响。方法通过组织切片的免疫荧光观察、RT-PCR、Q-PCR、Western blot等方法检测毛囊生长期[出生后8 d(P8d)]、毛囊退化期(P16d)、毛囊静止期(P25d)的Gsdma3基因突变鼠(AE鼠)和野生型鼠(WT鼠)背部皮肤Krt6b基因的表达情况。利用免疫荧光进一步观察注射有Gsdma3和RFP腺病毒的Gsdma3基因突变鼠和野生型鼠的Krt6b基因的表达情况。结果 Gsdma3基因突变鼠的Krt6b基因表达量在各个时相点均高于野生型鼠(P<0.05)。其中,在出生后16 d Krt6b基因表达量达到最高,而在出生后25 d降至最低。对Gsdma3基因突变鼠进行基因回补后,免疫荧光、实时定量PCR及Western blot检测结果显示Krt6b基因表达减弱。同时,对野生型鼠过表达Gsdma3基因后,免疫荧光、实时定量PCR及Western blot结果显示Krt6b基因表达量降低。结论小鼠皮肤角质形成细胞中,Krt6b基因的表达受Gsdma3基因抑制。

Objective To determine the effect of Gsdermin A3（ Gsdma3） on the expression of Krt6 b.Methods Q-PCR,RT-PCR,Western blotting and immunohistochemical staining were used to detect the expression of Krt6 b at mRNA and protein levels in Gsdma3 mutant mice（ AE） and C57 BL /6J wild-type mice（ WT） at 3 time points in mouse hair follicle cycle（ anagen： 8 d postnatally,catagen： 16 d postnatally,telogen： 25 d postnatally）. Immunofluorescence assay was also used to determine the effect of the adenoviruses carrying Gsdma3 gene（ Ad Gsdma3） or control gene（ AdRFP） on the expression of Krt6 b in AE and WT mice. Results The expression level of Krt6 b was obviously higher in Gsdma3 mutant mice than WT mice（ P〈0. 05）. At P16,the expression of Krt6 b reached the highest level,while on P25 it reduced to the lowest level. Immunofluorescence assay,real-time PCR and Western blotting showed that the expression of Krt6 b was decreased in Gsdma3 mutant mice after Gsdma3 expression rescue. Meanwhile,immunofluorescence assay showed that the expression of Krt6 b was also decreased in WT mice with Gsdma3 over-expression.Conclusion Gsdma3 inhibits the expression of Krt6 b in mouse keratinocytes.