[Prediction of anti-gastric cancer effect of Panacis Quinquefolii Radix by network pharmacology and in vivo validation]

This study investigated the anti-gastric cancer activity and mechanism of Panacis Quinquefolii Radix(Panax quinquefolium L.), and preliminarily compared the in vivo anti-gastric cancer efficacy of American-imported(JK-AG) and domestically produced(Shandong) Panacis Quinquefolii Radix decoctions(SD-AG). Based on network pharmacology predictions, a LUC-MGC803 cell ectopic gastric cancer nude mouse model was established. Mice were administered JK-AG and SD-AG at 1 g·kg~(-1) via gavage for 21 consecutive days. The positive control group received intraperitoneal injections of 5-fluorouracil(5-FU) at 20 mg·kg~(-1) every other day. Tumor volume was measured during the experiment. At the end of the experiment, high-resolution ultrasound imaging was used to measure tumor size, and an in vivo imaging system was used to observe tumor fluorescence expression. Tumor tissues were excised, weighed, and subjected to pathological histological examination. The terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL) assay and TUNEL+caspase-1 fluorescence double staining were used to detect tissue apoptosis and caspase-1 expression. Immunohistochemistry(IHC), Western blot(WB), and quantitative real-time PCR(RT-qPCR) techniques were used to detect the expression of predicted key proteins and genes involved in apoptosis and pyroptosis in tumor tissues. Network pharmacology predictions indicated that Panacis Quinquefolii Radix had anti-gastric cancer activity, primarily due to its ginsenoside components. Apoptosis induced by tumor necrosis factor(TNF)-α activation and caspase-1 may be key molecular mechanisms of its anti-cancer effects. In in vivo anti-tumor studies, compared to the model group, JK-AG and SD-AG did not affect the body weight or organ indices of the mice, whereas 5-FU significantly reduced body weight(P<0.05) and increased lung and liver indices(P<0.05). The tumor inhibition rates of JK-AG and SD-AG were 29.76% and 27.97%, respectively, which were lower than that of 5-FU. However, both JK-AG and SD-AG significantly reduced tumor tissue fluorescence intensity and three-dimensional tumor volume(P<0.05 or P<0.01). HE staining results showed that the drug groups had larger areas of tumor tissue necrosis. TUNEL and TUNEL+caspase-1 fluorescence staining indicated that both groups induced tumor cell apoptosis and increased caspase-1 expression. Both JK-AG and SD-AG significantly increased the expression of cleaved-caspase-8 and cleaved-caspase-3 proteins in tumor tissues detected by IHC and WB, as well as the mRNA expression of TNF-α, caspase-8, and caspase-9 detected by PCR, while reducing B-cell lymphoma 2(Bcl-2) protein expression. IHC showed that SD-AG increased the expression of TNF-α, caspase-9, cleaved-caspase-9, and tumor necrosis factor receptor 1(TNFR1) proteins more significantly than JK-AG, and also increased the Bax/Bcl-2 ratio, whereas JK-AG increased Bax mRNA expression more significantly than SD-AG. Additionally, JK-AG and SD-AG significantly increased the expression of NLRP3, caspase-1, cleaved-caspase-1, Gasdermin D(GSDMD), cleaved-GSDMD, IL-18, Gasdermin E(GSDME), and GSDME-NT proteins and the mRNA expression of caspase-1 and GSDME. JK-AG increased cleaved-caspase-1 protein expression more significantly than SD-AG in WB analysis. At the same time, JK-AG increased the expression of GSDMD mRNA, which was significantly higher than that of SD-AG group. For GSDME-NT protein expression, SD-AG increased it more significantly than JK-AG. The above studies indicate that Panacis Quinquefolii Radix has anti-gastric cancer effects, and the efficacy of American and Shandong ginsengs is comparable. Their mechanisms are related to TNF-α-mediated extrinsic apoptosis pathways and the induction of pyroptosis. The sensitivity of their action targets varies between American and Shandong ginsengs.