基于网络药理学探讨西黄解毒胶囊抑制三阴性乳腺癌生长与转移的机制及实验验证  

Exploring the Mechanism of Xihuang Jiedu Capsule in Inhibiting the Growth and Metastasis of Triple-Negative Breast Cancer Based on Network Pharmacology and Experimental Verification

作　　者：桂越蓉 王烁 董军 王昀 侯炜[2] GUI Yue-rong;WANG Shuo;DONG Jun;WANG Yun;HOU Wei(Shunyi Hospital,Beijing Traditional Chinese Medicine Hospital,Bejjing 101300;Guang'anmen Hospital,China A-cademy of Chinese Medical Sciences,Beijing 100053;Chongqing University Cancer Hospital,Chongqing 400030)

机构地区：[1]北京中医医院顺义医院,北京101300 [2]中国中医科学院广安门医院,北京100053 [3]重庆大学附属肿瘤医院,重庆400030

出　　处：《世界中西医结合杂志》2024年第9期1773-1782,1795,共11页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金(82074239)。

摘　　要：目的通过网络药理学的方法探索西黄解毒胶囊抗三阴性乳腺癌(TNBC)的机制,并通过体内和体外实验进行验证。方法通过检索中草药系统药理学平台(TCMSP)数据库和查阅文献,获得西黄解毒胶囊中所含药物的有效活性成分,并在靶点筛选栏和SwissTargetPrediction数据库筛选活性成分对应的人类基因靶点。将药物靶点与在GEO数据库(GSE38959)中获得的TNBC差异表达基因相映射取交集,获得西黄解毒胶囊治疗TNBC的靶点,将交集靶点导入String在线数据库构建PPI网络,根据度值筛选西黄解毒胶囊治疗TNBC的核心靶点,在DAVID数据库对西黄解毒胶囊治疗TNBC的核心靶点进行GO和KEGG富集分析,以探索西黄解毒胶囊治疗TNBC的可能机制。在体内和体外分别采用蛋白免疫印迹和荧光定量PCR实验检测信号通路上关键信号分子的表达情况。结果网络药理学部分获得西黄解毒胶囊抗TNBC的核心靶点有HIF1A、VEGFA等,生物过程主要涉及信号转导、转录调控、增殖调节等,参与的信号通路主要有癌症信号通路、PI3K-Akt信号通路、NF-κB信号通路等。实验部分发现西黄解毒胶囊能抑制4T1细胞和移植瘤中EGFR、AKT(pan)、p-AKT(Ser473)、NF-κBp65、p-NF-κBp65、PI3Kp85、PI3Kp110α蛋白的表达,基因层面荧光定量PCR试验得出一致得结果。结论西黄解毒胶囊可能通过抑制PI3K/AKT/NF-κB信号通路发挥抗TNBC的作用。Objective The mechanism of Xihuang Jiedu capsule against triple-negative breast cancer(TNBC)was explored using a network pharmacology approach,with in vivo and in vitro experiments conducted for verification.Methods The active ingredients of Xihuang Jiedu capsule were identified through the Traditional Chinese Medicine Systems Pharmacology Database(TCMSP)and literature review.Corresponding human gene targets of these active ingredients were obtained via the target screening section and the SwissTargetPrediction database.The drug targets were intersected with differentially expressed genes(DEGs)of TNBC obtained from the GEO database(GSE38959)to identify potential targets of Xihuang Jiedu capsule in treating TNBC.These intersecting targets were imported into the STRING online database to construct a PPI network,and core targets were identified based on degree values.GO and KEGG enrichment analyses of these core targets were performed using the DAVID database to explore the possible mechanisms of Xihuang Jiedu capsule in treating TNBC.Western blotting and quantitative real-time PCR(qRT-PCR)were used to detect the expression of key signaling molecules in the signaling pathway in vivo and in vitro.Results Network pharmacology identified core targets of Xihuang Jiedu capsule against TNBC,including HIF1A,VEGFA.The Biological processes mainly involved signal transduction,transcriptional regulation,and proliferation regulation,with participation in pathways such as the cancer signaling pathway,PI3K-Akt signaling pathway,and NF-κB signaling pathway.The experimental results showed that Xihuang Jiedu capsule inhibited the expression of EGFR,AKT(pan),p-AKT(Ser473),NF-κBp65,p-NF-κBp65,PI3Kp85,and PI3Kp110αproteins in 4T1 cells and transplanted tumors.Consistent results were obtained at the genetic level through qRT-PCR experiments.Conclusion Xihuang Jiedu capsule may exert its anti-TNBC effects by inhibiting the PI3K/AKT/NF-κB signaling pathway.

关 键 词：三阴性乳腺癌 西黄解毒胶囊 网络药理学 机制 细胞 动物 

分 类 号：R285.6[医药卫生—中药学]