摘要
目的:采用网络药理学和体外细胞实验研究枇杷清肺饮通过MAPK/NF-κB信号通路调控LPS诱导的HaCaT细胞炎性损伤机制。方法:通过TCMSP数据库、GeneCards、OMIM和TTD数据库,取枇杷清肺饮和痤疮交集靶点,使用STRING数据库建立PPI网络,DAVID数据库筛选靶点GO功能并进行KEGG通路富集分析,Cytoscape软件构建“活性成分-靶点-KEGG-痤疮”网络,预测其作用机制。进一步采用细胞实验验证预测靶点的准确性,采用LPS诱导HaCaT细胞炎症模型,CCK-8检测不同浓度枇杷清肺饮含药血清对细胞活力的影响,qPCR和ELISA检测HaCaT细胞IL-1β、AKR1B1、IL-4、TNF-α的mRNA及蛋白表达水平,Western blot检测NF-κBp65、P38-MAPK和JNK-MAPK蛋白的表达水平。结果:通过网络分析可知,6种中药包含153个活性成分共304个靶标,与痤疮共有靶标基因118个,关键靶点涉及IL-1β、IL-4、TNF-α、AKR1B1等;GO功能富集分析得到条目1 922个(P<0.05);KEGG通路富集分析筛选到131条信号通路,其中有4条通路与炎症相关,分别为PI3K/Akt、MAPK、TNF信号通路和IL-17信号通路。体外细胞实验结果显示,枇杷清肺饮能抑制HaCaT细胞炎性损伤模型中IL-1β、IL-4、TNF-α、AKR1B1 mRNA及蛋白的表达,抑制P38-MAPK和JNK-MAPK的磷酸化以及NF-κBp65向细胞核内的转录。结论:枇杷清肺饮具有明显的抗炎作用,其机制可能与其通过作用于IL-1β、IL-4、TNF-α、AKR1B1等靶点调节NF-κB和MAPK信号通路有关。
Objective:Network pharmacology and in vitro cell experiment were used to study the mechanism of PIPA Qingfei Yin regulating LPS induced inflammatory injury of HaCaT cells through MAPK/NF-κB signal pathway.Methods:Selected the cross targets of PIPA Qingfei Yin and acne through TCMSP database,GeneCards,OMIM and TTD database,STRING database was used to establish PPI network,analyzed targets GO function and KEGG pathway enrichment by DAVID Database,used Cytoscape to construct network of "active ingredients-btargets-KEGG-acne" and to predict its mechanism of action.Accuracy of targets prediction was further verified by cell experiment,inflammation model of HaCaT cells was induced by LPS,detected effect of different concentration of PIPA Qingfei Yin on cell viability by using CCK-8,expressions of IL-1β,AKR1B1,IL-4,TNF-α mRNA and protein in HaCaT cells were detected by qPCR and ELISA,and expressions of NF-κBp65,P38-MAPK and JNK-MAPK were detected by Western blot.Results:Through network analysis,it was found that 6 traditional Chinese medicines contain 153 active ingredients and 304 targets,and there were 118 target genes with acne.Key targets involved in IL-1β,IL-4,TNF-α,AKR1B1,etc.GO enrichment analysis resulted in1 922 items(P<0.05),131 signal pathways were screened by KEGG pathway enrichment analysis,and 4 of them were related to inflammation,including PI3K/Akt,MAPK,TNF signal pathway and IL-17 signal pathway.Results of in vitro cell experiments show that PIPA Qingfei Yin inhibited expressions of IL-1β,IL-4,TNF-α,AKR1B1 mRNA and protein,phosphorylation of P38-MAPK and JNK-MAPK,and transcription of NF-κBp65 into nucleus of cells ininflammatory model of HaCaT cells.Conclusion:PIPA Qingfei Yin has obvious anti-inflammatory effect,which mechanism may be related to the regulation of NF-κB and MAPK signal pathway by targeting IL-1β,IL-4,TNF-α and AKR1B1.
作者
韩晓晴
吴景东(指导)
侯殿东
张小卿
王靓怡
HAN Xiaoqing;WU Jingdong;HOU Diandong;ZHANG Xiaoqing;WANG Liangyi(Liaoning University of Traditional Chinese Medicine,Shenyang 110000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第10期1189-1195,共7页
Chinese Journal of Immunology
基金
国家自然科学基金面上项目(81774023)。
