Integrated Network Pharmacology and Serum Metabolomics Analysis to Reveal the Mechanism of Gypenosides Against Lung Cancer and Enhancing Cisplatin Efficiency in Lewis Lung Cancer Mice

Abstract

Ethnopharmacological relevance: It is reported that gypenosides (GYP) have anticancer activity against various cancers. The purpose of this study was to investigate the anticancer effect and mechanism of GYP enhancing cisplatin efficacy on Lewis tumor-bearing mice. Cisplatin is a kind of chemotherapy drug for lung cancer.

Aim of the study: Metabolomics and network pharmacology methods were used to study the potential mechanism of GYP in treating lung cancer (LC) and enhancing cisplatin anticancer activity.Materials and methodsMTT assay was used to detect the effect of GYP on the viability of LLC cells and BEAS-2B cells; The anti-tumor effect of GYP was evaluated by the weight, volume and histopathological examination of tumor tissue; UPLC-QTOF/MS untargeted metabolomics technique combined with network pharmacology technique was used to study the anti-tumor effect of GYP on Lewis lung carcinoma (LLC) mice and the mechanism of GYP enhancing anti-lung cancer, and western blot was used to verify the related proteins and pathways.

Results: MTT assay showed that GYP could inhibit the proliferation of LLC cells in a dose-dependent manner. The sarcoma volume and weight of LLC tumor bearing mice in GYP administration groups, cisplatin group and GYP + cisplatin groups were significantly lower than those in model group, and the sarcoma volume and weight of LLC mice in GYP + cisplatin groups were significantly lower than those in GYP administration groups and cisplatin group. Histopathological analysis of the tumors showed that GYP and/or cisplatin had an ameliorative effect on lung cancer, with the best anticancer effect in the GYP + cisplatin groups. Serum metabolomics identified 57 biomarkers related to LLC mice, of which 23 biomarkers were related to the therapeutic effect of GYP on lung cancer. The therapeutic effects of GYP on LLC mice mainly involve alpha-linolenic acid metabolism, glutathione metabolism, sphingolipid metabolism, glycerophospholipid metabolism, tryptophan metabolism, and primary bile acid biosynthesis. The results of metabolomics combined with pharmacological techniques and western blotting showed that GYP could indirectly affect five metabolites through MAPK14, STAT3, TYMS, and EGFR to affect three metabolic pathways, and GYP may exert an anticancer effect and enhance the anti-lung cancer effect through MAPK14/STAT3 signaling pathway.

Conclusion: GYP has anti-tumor effect and enhances the anti-cancer effect of the chemotherapy drug cisplatin. Metabolomics and network pharmacology combined with western blot further verify the mechanism of GYP anti-cancer and GYP enhancing the anti-cancer effect of cisplatin through the MAPK14/STAT3 signaling pathway.

Note:

Funding Information: This work was supported financially by the National Natural Science Foundation of China (No. 81673692); National Key R&D Program of China (N0. 2017YFC1704000); The Doctoral Student’s Independent Research Project of Minzu University of China (No. BBZZKY 2020041).

Declaration of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethics Approval Statement: All animal protocols were approved by the Biological and Medical Ethics Committee of China University for Nationalities (ECMUC201903AO, 9 March 2019), and all animal work was performed in accordance with relevant guidelines and regulations.

Keywords: Gynostemma pentaphyllum, gypenosides, cisplatin, Metabolomics, Network pharmacology, Lung cancer