大黄素联合紫杉醇治疗非小细胞肺癌的作用机制探讨

The mechanistic exploration of the combined treatment of rhodopsin and paclitaxel in non-small cell lung cancer

目的 基于网络药理学探讨大黄素联合紫杉醇治疗非小细胞肺癌(NSCLC)的作用机制,并应用实验验证网络药理学结果。方法 通过中药系统药理学分析平台TCMSP以及医药信息查询库药智数据查找大黄素和紫杉醇的相关靶点,人类基因数据库GeneCards查找NSCLC的相关疾病靶点。应用STRING数据库和Cytoscape构建大黄素联合紫杉醇成分-靶点网络,分析大黄素联合紫杉醇治疗NSCLC的作用靶点、生物学过程以及相关通路,同时利用MTT实验检测大黄素联合紫杉醇对A549细胞增殖的抑制率,并ELISA检测网络药理学得到的核心靶基因水平。结果 大黄素联合紫杉醇与NSCLC的共同靶点基因32个,PPI分析获得前十个靶点蛋白为TP53、CASP3、TNF、EGF、PTGS2、MYC、KDR、TGFB1、MMP9、PPARG,GO分析与KEGG通路分析获得大黄素联合紫杉醇可能作用于NSCLC的150个生物学过程和45条相关通路。MTT结果表明大黄素联合紫杉醇对A549细胞具有明显生长抑制作用(P<0.05),且联合组抑制率明显高于大黄素及紫杉醇单用组(P<0.05)。ELISA结果表明,大黄素与紫杉醇联合组能明显增加A549细胞CASP3含量(P<0.05),降低TNF-α含量(P<0.05),其中联合组效果明显优于大黄素组和紫杉醇组(P<0.05)。结论 大黄素联合紫杉醇能够通过调节细胞增殖与凋亡、调控肿瘤抑制基因表达、炎症反应的相关靶点通路对NSCLC起到治疗作用,大黄素联合紫杉醇有抑制A549细胞增殖的作用,其机制可能与调控CASP3、TNF-α的水平相关。

Objective To explore the mechanism of action of rhodopsin combined with paclitaxel in the treatment of non-small cell lung cancer(NSCLC)based on network pharmacology,and to apply experiments to verify the network pharmacology results.Methods The TCMSP,a systematic pharmacology analysis platform for traditional Chinese medicine,and the Pharmaceutical Information Query Library(PIQ),Pharmaceutical Intelligence Data(PID)were used to find out the relevant targets of rhodopsin and paclitaxel,and GeneCards,a human gene database,was used to find out the relevant disease targets of NSCLC.The STRING database and Cytoscape were used to construct the constituent-target network of rhodopsin combined with paclitaxel,and to analyze the targets,biological processes,and related pathways of rhodopsin combined with paclitaxel in the treatment of NSCLC,and at the same time,to detect the inhibitory rate of rhodopsin combined with paclitaxel on the proliferation of A549 cells using the MTT assay,and to detect the levels of core target genes obtained by ELISA for the pharmacological aspects of the network.Results The common target genes of rhodopsin combined with paclitaxel and NSCLC were 32,and the top ten target proteins obtained by PPI analysis were TP53,CASP3,TNF,EGF,PTGS2,MYC,KDR,TGFB1,MMP9,and PPARG.GO analysis and KEGG pathway analysis identified 150 biological processes and 45 relevant pathways that rhodopsin combined with paclitaxel might act on in NSCLC.MTT results showed that rhodopsin combined with paclitaxel had a significant growth inhibitory effect on A549 cells(P<0.05),and the inhibition rate of the combined group was significantly higher than that of the rhodopsin and paclitaxel alone group(P<0.05).ELISA results showed that the combined group of rhodopsin and paclitaxel significantly increased the CASP3 content of A549 cells(P<0.05),and decreased TNF-αcontent(P<0.05),in which the effect of the combined group was significantly better than that of the rhodopsin and paclitaxel groups(P<0.05).Conclusion The combination of rhodopsin and paclitaxel can play a therapeutic role in NSCLC by regulating cell proliferation and apoptosis,modulating the expression of tumor suppressor genes,and inflammatory response of the relevant target pathways,and the combination of rhodopsin and paclitaxel has an inhibitory effect on the proliferation of A549 cells,and the mechanism of this may be related to the modulation of the levels of CASP3 and TNF-α.