摘要
基于网络药理学及急性肺损伤(acute lung injury,ALI)体外模型方法探索金荞麦及其主要活性成分治疗急性肺损伤的作用机制。通过TCMSP数据库,结合口服生物利用度、类药性作为限定条件获取金荞麦主要活性成分。检索PubChem数据库获得金荞麦活性成分相关靶蛋白,在GeneCards数据库收集与ALI相关靶基因,应用STRING 11.0构建化合物靶蛋白-ALI靶基因相互作用(PPI)网络,借助IPA生物网络分析平台对金荞麦潜在化合物靶蛋白与ALI靶基因共同作用的通路进行比较分析,预测金荞麦治疗ALI的潜在关键靶点和信号通路。最后,通过金荞麦活性成分表儿茶素(epicatechin,EC)干预脂多糖诱导的RAW264.7细胞进行体外实验,对潜在关键靶点及通路进行验证。网络药理学结果显示,表儿茶素、原花青素B1、木犀草素等15种潜在活性成分可能通过RELA(P65)等关键靶点,作用于核因子-κB(nuclear transcription factor-κB,NF-κB)信号通路、转化生长因子-β(transforming growth factor-β,TGF-β)信号通路、腺苷酸激活蛋白激酶[adenosine 5'-monophosphate(AMP)-activated protein kinase,AMPK]信号通路等发挥治疗ALI作用。体外实验结果显示,25μmol·^(L-1)的EC与细胞共同培养24 h时与对照组相比细胞存活率差异无统计学意义,其发挥抗炎作用主要通过抑制NF-κB信号通路中p65磷酸化蛋白的表达水平,从而下调其下游促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、一氧化氮(nitric oxide,NO)表达,上调抑炎因子白细胞介素-10(interleukin-10,IL-10)表达。以上结果表明,金荞麦主要通过抑制NF-κB信号通路中p65蛋白的磷酸化,进而下调信号通路下游促炎细胞因子发挥治疗ALI作用。
The present study explored the mechanism of Fagopyri Dibotryis Rhizoma(FDR)and its main active components in the treatment of acute lung injury(ALI)based on the network pharmacology and the in vitro experiments.The main active components of FDR were obtained from the TCMSP database and screened by oral bioavailability and drug-likeness.The related target proteins of FDR were retrieved from the PubChem database,and the target genes related to ALI were screened out from the GeneCards database.A protein-protein interaction(PPI)network of compound target proteins and ALI target genes was constructed using STRING 11.0.Ingenuity Pathway Analysis(IPA)platform was used to analyze the common pathways of the potential compound target proteins of FDR and ALI target genes,thereby predicting the key targets and potential signaling pathways of FDR for the treatment of ALI.Finally,the potential pathways and key targets were verified by the in vitro experiments of lipopolysaccharide-induced RAW264.7 cells intervened by epicatechin(EC),the active component of FDR.The results of network pharmacology showed that 15 potential active components such as EC,procyanidin B1,and luteolin presumedly functioned in the treatment of ALI through nuclear transcription factor-κB(NF-κB)signaling pathway,transforming growth factor-β(TGF-β)signaling pathway,and adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)signaling pathway through key targets,such as RELA(P65).The results of in vitro experiments showed that 25μmol·L^(-1) EC had no toxicity to cells and could inhibit the expression of the p65-phosphorylated protein in the NF-κB signaling pathway to down-regulate the expression of downstream inflammatory cytokines,including tumor necrosis factor-α(TNF-α),IL-1βand nitric oxide(NO),and up-regulate the expression of IL-10.These results suggested that the therapeutic efficacy of FDR on ALI was achieved by inhibiting the phosphorylation of p65 protein in the NF-κB signaling pathway and down-regulating the level of proinflammatory cytokines downstream of the signaling pathways.
作者
耿琦
刘斌
赵鹏程
熊一白
李立
易剑峰
吕诚
GENG Qi;LIU Bin;ZHAO Peng-cheng;XIONG Yi-bai;LI Li;YI Jian-feng;LYU Cheng(College of Chemical&Biology Engineering,Yichun University,Yichun 336000,China;Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Life Sciences,Northwestern Polytechnical University,Xi'an 710000,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2021年第18期4816-4823,共8页
China Journal of Chinese Materia Medica
基金
国家“重大新药创制”科技重大专项(2018ZX10101001-005)
中央级公益性科研院所基本科研业务费专项(ZZ13-024-1,Z0653)。
关键词
金荞麦
急性肺损伤
网络药理学
分子机制
Fagopyri Dibotryis Rhizoma
acute lung injury
network pharmacology
molecular mechanism
