摘要
利用中药网络药理学结合脂多糖(LPS)诱导RAW 264.7细胞建立的炎症模型,共同探讨铁苋菜(AAL)治疗溃疡性结肠炎(UC)的作用机制。基于文献研究并结合OMIM、Drugbank、TDD数据库筛选得到AAL以及UC相关靶标,进行GO功能分析和KEGG通路富集分析。将RAW 264.7细胞分为空白对照组、LPS组、阳性药物组、铁苋菜高、中、低剂量组(900,800,700 mg/L)。观察高、中、低剂量AAL对RAW 264.7的细胞活力和细胞凋亡的影响,同时采用qRT-PCR和Western blot检测高、中、低剂量AAL对NLRP3、GSDMD、Caspase-1、ASC、IL-1β、IL-18、MUC-2、OCLN的mRNA与蛋白表达水平的影响。网络药理学结果显示AAL治疗UC主要涉及凋亡过程的负调控及PI3K-Akt信号通路等过程。体外试验显示700,800,900 mg/L AAL显著抑制LPS处理的巨噬细胞的活力,减少细胞凋亡,降低NLRP3、GSDMD、Caspase-1和ASC mRNA和蛋白的表达。以上结果表明AAL可通过抑制NLRP3/Caspase-1信号通路,减少炎症因子IL-18和IL-1β的分泌,从而减轻LPS诱导的炎症,治疗UC。
The aim of this study is to use traditional Chinese medicine network pharmacology combined with lipopolysaccharide(LPS)induced RAW 264.7cell inflammation model to discuss molecular mechanism of Acalypha australis L.(AAL)in the treatment of ulcerative colitis(UC).Literature research combined with OMIM,Drugbank and TDD databases,the UC-related targets of AAL were obtained.Then GO function analysis and KEGG pathway enrichment analysis were carried out.RAW 264.7cells were divided into blank control group,LPS group,positive drug group,and high,medium and low dose groups of AAL(900,800,700mg/L).The effects of high,medium and low doses of AAL on the cell viability and apoptosis of RAW 264.7were observed.At the same time,qRT-PCR and Western blot were used to detect the effects of different doses of AAL on NLRP3,GSDMD,Caspase-1,ASC,IL-1β,IL-18,MUC-2and OCLN mRNA and protein expression levels.The results of network pharmacology showed that AAL treatment of UC mainly involved the negative regulation of apoptosis and PI3K-Akt signaling pathway.In vitro experiments showed that 700,800,and 900mg/L of AAL significantly inhibited LPS-stimulated macrophage viability,decreased cell apoptosis,and decreased the mRNA and protein expressions of NLRP3,GSDMD,Caspase-1and ASC.The above results indicate that AAL can inhibit the NLRP3/Caspase-1signaling pathway and reduce the secretion of inflammatory factors IL-18and IL-1β,thereby reducing LPS-induced inflammation and treating ulcerative colitis.
作者
尕玉
樊艺萌
王惠茹
魏媛媛
张艳楠
郝智慧
GA Yu;FAN Yimeng;WANG Huiru;WEI Yuanyuan;ZHANG Yannan;HAO Zhihui(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第5期1019-1028,共10页
Chinese Journal of Veterinary Science
基金
国家自然科学基金面上资助项目(32172897)
中央本级重大增减支资助项目(名贵中药资源可持续利用能力建设项目(2060302))
作者简介
尕玉(1998-),回族,女,硕士研究生;通讯作者:郝智慧,E-mail:haozhihui@cau.edu.cn
