基于网络药理学和实验研究Jolkinolide B治疗结肠肿瘤的作用机制

Mechanism of jolkinolide B in treatment of carcinoma of colon based on network pharmacology and experiment

通过网络药理学方法及生物学实验研究Jolkinolide B治疗结肠肿瘤的作用机制。通过TCMSP等数据库预测Jolkinolide B作用靶点,并通过靶点富集分析及分子对接预测其防治结肠肿瘤的作用机制,基于预测结果,体内生物学实验初步验证Jolkinolide B对移植瘤小鼠肿瘤的抑制作用,MTT法检测不同浓度的Jolkinolide B对HCT116细胞的增殖抑制活性,流式细胞术检测细胞凋亡情况,细胞划痕实验观察其对HCT116细胞迁移能力的影响。Western blot检测相关凋亡蛋白Bax、Bcl-2、金属基质蛋白酶MMP3和MMP12的表达。网络靶点分析结果显示Jolkinolide B与结肠癌(CRC)匹配后得到78个靶点,富集信号通路主要有IL-17信号通路、PD-1/PD-L1信号通路等。分子对接结果显示Jolkinolide B以氢键与STAT3、MAPK8、JUN核心蛋白结合,结合构象较稳定。体内实验结果表明,Jolkinolide B能有效抑制移植瘤小鼠肿瘤体积;细胞水平验证实验结果表明,Jolkinolide B能够有效地抑制结肠肿瘤细胞HCT116的增殖,促进细胞的凋亡,抑制细胞的迁移;Western blot结果表明Jolkinolide B明显上调Bax/Bcl-2蛋白的表达,诱导了HCT116细胞的凋亡;同时,Jolkinolide B明显下调金属基质蛋白酶MMP3、MMP12的表达,抑制肿瘤细胞侵袭和转移。综上,Jolkinolide B可通过多靶点、多途径实现干预CRC的作用,其抗肿瘤活性可能与影响凋亡基因Bax及Bcl-2、金属基质蛋白酶MMP3、MMP12的表达有关。本研究为Jolkinolide B的临床用药和防治的作用机制研究提供了参考。

To explore the mechanism of jolkinolide B in the treatment of carcinoma of colon based on network pharmacology and in vivo/vitro experiment.The Target of jolkinolide B was predicted by TCMSP and other databases,and the mechanism of jolkinolide B against carcinoma of colon(CRC)was analyzed by Target enrichment analysis and molecular docking.Based on the prediction results,in vivo experiment was used to observe the inhibitory effects of jolkinolide B on the growth of transplanted tumor.The MTT assay was used to detect the proliferation inhibitory activity of jolkinolide B at different concentrations on HCT116 cells,flow cytometry was used to detect cell apoptosis,and a cell scratch assay was used to observe the effects of jolkinolide B on migration ability of HCT116 cells.Western blot was used to detect the expression of related apoptotic proteins Bax,Bcl-2,MMP3 and MMP12.In network target analysis,jolkinolide B matched with CRC to get 78 targets.Enrichment signal pathways mainly include IL-17 signal pathway,PD-1/PD-L1 signal pathway,etc.Molecular docking results showed that jolkinolide B bonded to STAT3,MAPK8 and Jun core proteins by hydrogen bonds,and the binding conformation was relatively stable.In vivo experiment showed that jolkinolide B obviously reduce the volume of the transplanted tumor.The results of cell-level verification experiments showed that jolkinolide B significantly inhibited the proliferation of HCT116 cells,promoted cell apoptosis,and inhibited cell migration;Western blot results showed that jolkinolide B significantly up-regulates the expression of Bax/Bcl-2,which induces cell apoptosis,and down-regulates the expression of MMP3 and MMP12,which inhibit tumor cell invasion and metastasis.This study provides a reference for the study on the clinical drug use and the mechanism of action of jolkinolide B.