中间丝相关蛋白的敲低影响正常人表皮角质形成细胞中的丝裂原活化蛋白激酶信号通路。
Knock-down of filaggrin influences the mitogen-activated protein kinases signaling pathway in normal human epidermal keratinocytes.
作者信息
Wang Shumei, Qiu Liyun, Meng Xianguang, Dang Ningning
机构信息
Department of Community Medicine, Jinan Central Hospital affiliated to Shandong University, No.105 Jiefang Road, Jinan 250013, Shandong Province, China.
Department of Pharmacy, Jinan Central Hospital affiliated to Shandong University, No.105 Jiefang Road, Jinan 250013, Shandong Province, China.
出版信息
Med Sci (Paris). 2018 Oct;34 Focus issue F1:94-98. doi: 10.1051/medsci/201834f116. Epub 2018 Nov 7.
BACKGROUND
Filaggrin is an essential structural protein of the stratum corneum binding to the keratin intermediate filaments to form a dense protein-lipid matrix. However, the function of filaggrin in epidermal terminal differentiation is not completely understood.
AIM
To evaluate the effects of filaggrin on normal human epidermal keratinocytes (NHEKs) and to investigate the relevant mechanisms.
METHODS
Short hairpin RNA (shRNA) technology was used to knock-down filaggrin in normal human epidermal keratinocytes (NHEKs). Western blot and real-time quantitative PCR (qRT-PCR) were performed to detect expression of filaggrin, differentiation-related proteins and MAPK-related proteins.
RESULTS
Filaggrin was successfully knocked down in NHEKs (99% efficiency). We found that the lack of filaggrin significantly decreased the expression of some differentiation-related proteins, including Cytokeratin 5 protein, Cytokeratin 14 protein, ST14 protein and SPRR3 protein (P<0.05). In addition, filaggrin knock-down significantly decreased expression of p-p38, p-ERK1/2, p-JNK, p-Akt, and p-NF-κB in NHEKs.
CONCLUSION
Our study shows that filaggrin regulates epidermal terminal differentiation and impairs MAPK signaling pathway in normal human epidermal keratinocytes.
背景
丝聚合蛋白是角质层的一种重要结构蛋白，它与角蛋白中间丝结合形成致密的蛋白质-脂质基质。然而，丝聚合蛋白在表皮终末分化中的功能尚未完全明确。
目的
评估丝聚合蛋白对正常人表皮角质形成细胞（NHEKs）的影响，并探讨相关机制。
方法
采用短发夹RNA（shRNA）技术敲低正常人表皮角质形成细胞（NHEKs）中的丝聚合蛋白。通过蛋白质免疫印迹法（Western blot）和实时定量聚合酶链反应（qRT-PCR）检测丝聚合蛋白、分化相关蛋白和丝裂原活化蛋白激酶（MAPK）相关蛋白的表达。
结果
NHEKs中的丝聚合蛋白成功被敲低（敲低效率为99%）。我们发现，丝聚合蛋白的缺失显著降低了一些分化相关蛋白的表达，包括细胞角蛋白5蛋白、细胞角蛋白14蛋白、ST14蛋白和富含脯氨酸的小蛋白3（SPRR3）蛋白（P<0.05）。此外，敲低丝聚合蛋白显著降低了NHEKs中磷酸化p38、磷酸化细胞外信号调节激酶1/2（ERK1/2）、磷酸化c-Jun氨基末端激酶（JNK）、磷酸化蛋白激酶B（Akt）和磷酸化核因子κB（NF-κB）的表达。
结论
我们的研究表明，丝聚合蛋白在正常人表皮角质形成细胞中调节表皮终末分化并损害MAPK信号通路。
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