Exploring the Mechanism of Action of Xianfang Huoming Decoction in Inhibiting Triple-Negative Breast Cancer and the Action of Its Principal Medicine, Lonicera japonica
LU Dongyu1, SUN Boyu1, WANG Rui1, ZHAO Run1, WANG Dawei1, LAN Tian1,2, TSENG Yiider1,2*
This study was to explore the mechanism of action of the Traditional Chinese Medicine formula XHD (Xianfang huoming decoction) in inhibiting TNBC (triple-negative breast cancer) and the action of its prin cipal medicine, Lonicera japonica. By using metabolomics combined with network pharmacology to analyze the differential components in the drug-containing serum of rats prepared with XHD (Xianfang group), XHD without Lonicera japonica (Jianjin group), Lonicera japonica decoction (Lonicera japonica group), and by searching for the targets, the intersection of the targets corresponding to the differential components of the three groups with the tar gets of the differential gene expression in TNBC was conducted to screen the core targets, and pathway enrichment was performed. Then, the inhibitory effect of XHD and Lonicera japonica on tumor growth and their regulatory effects on key pathways were verified: a TNBC in situ transplanted tumor model was constructed in mice for drug intervention. The groups included the normal group, the model group, the Xianfang group, the Jianjin group, the Lonicera japonica group, and the positive drug group (cisplatin group). Animal experiments were conducted. The MDA-MB-231 cells were used as the carrier for drug addition, and the experiments were carried out in the Con trol group, the Xianfang group, the Jianjin group, and the Lonicera japonica group. LC-MS analysis revealed that the drug-containing serum samples from the Xianfang group, the Jianjin group, and the Lonicera japonica group identified 151, 131, and 65 different components respectively, corresponding to 1 338, 1 316, and 1 181 targets. After interacting with the TNBC targets, all of them were enriched in pathways such as Cell cycle, PI3K-AKT, and Apoptosis, indicating that XHD can affect TNBC through the apoptotic pathway. In vivo experiments found that the decoction of the Xianfang group, the Jianjin group, and the Lonicera japonica group had no obvious toxicity to the liver and kidneys of mice; at the same time, all of these groups could inhibit the growth of TNBC tumors. Immuno histochemical results showed that, compared with the model group, the levels of Bcl-xl expression in the Xianfang group and the Lonicera japonica group were significantly reduced. Western blot results showed that the expression of Caspase-3 and Bax was significantly increased in the Xianfang group and Lonicera japonica group, compared with the model group, and the expression of Bcl-xl was significantly reduced in the Xianfang group, Jianjin group, and Lonicera japonica group, compared with the model group. In vitro experiments found that, compared with the Control group, the drug-containing serum of the Xianfang group, Jianjin group, and Lonicera japonica group could significantly inhibit the activity of MDA-MB-231 cells. The Western blot results showed that compared with the Control group, the expression level of Caspase-3 in the Xianfang group significantly increased, while the expres sion levels of Bcl-xl in the Xianfang group, Jianjin group and Lonicera japonica group all significantly decreased. Through metabolomics and network pharmacology, and with the mutual verification of in vivo and in vitro experi ments, this study found that XHD and its principal medicine Lonicera japonica exerted anti-TNBC effects by regu lating the apoptotic pathway through multi-component, multi-target, inhibiting tumor growth in vivo and inhibiting the activity of MDA-MB-231 cells in vitro.
CN
EN