1.Study on the Mechanism of Triterpenoids from Inonotus Obliquus Against Gastric Cancer HeLa Cell Based on Network Pharmacology and Experimental Validation
Junjing SHAO ; Ying YANG ; Chuangong WANG ; Xiaoping ZHANG
Chinese Journal of Modern Applied Pharmacy 2024;41(3):313-323
OBJECTIVE
To explore the potential mechanism of anti-HeLa cell of triterpenoids from Inonotus obliquus by network pharmacology, molecular docking and in vivo verification experiments.
METHODS
Based on literature research and database, the main active components of triterpenoids from Inonotus obliquus were screened, and the active components and HeLa cell intersection targets were found. PPI network was constructed, GO and KEGG enrichment analysis were carried out, and molecular docking was verified. HeLa cell nude mice model was established, and Inonotus obliquus triterpenoids were administered by gavage. The indexes were detected and detected by Western blotting after euthanizing nude mice.
RESULTS
After screening, 274 associated targets of 15 qualified active components were obtained. Intersecting with 2 046 HeLa cell-related targets, 104 potential anti-HeLa cell targets of Inonotus obliquus triterpenoids were obtained. Through the "drug-component-target-disease" network analysis, PIK3CA, MAPK3, MDM2, AR and CCND1 might be potential targets for anti-HeLa cell of triterpenoids purified from Inonotus obliquus. The results of KEGG enrichment analysis showed that multiple signal pathways such as PI3K/Akt signal pathway, Ras signal pathway and MAPK signal pathway might be the potential anti-HeLa cell pathway of triterpenoids in Inonotus obliquus. The results of molecular docking showed that the core components and key targets had strong potential binding ability. The in vivo test results showed that Inonotus obliquus triterpenoids could inhibit tumor growth, regulate immune function, promote tumor apoptosis and inhibit PI3K and AKT phosphorylation in model mice.
CONCLUSION
Based on the methods of network pharmacology, molecular docking and in vivo verification, it is proved that the triterpenoids of Inonotus obliquus play the role of anti-HeLa cell through multi-components and multi-targets, which lays a foundation for further research and development of Inonotus obliquus and the discussion of its anti-tumor mechanism.
2.Study on the purification technology of total triterpenoid extracts from Inonotus obliquus and their anti -tumor activity in vitro
Junjing SHAO ; Ying YANG ; Dalong MA ; Zhiqiang LYU ; Jingzhen TIAN ; Xiaoping ZHANG
China Pharmacy 2022;33(18):2198-2203
OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus ,and to investigate the anti -tumor activity of its purified products . METHODS Using inotodiol as control ,the method was established for the content determination of total triterpenoid in I. obliquus. The type of macroporous adsorption resin ,sample volume ,sample concentration,sample flow rate ,eluent volume ,eluent dosage and elution flow rate were selected by single factor experiments . The purification technology of the crude extract was determined and verified . The effects of total triterpenoid purified from I. obliquus on the proliferation ,migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test , migration test ,flow cytometry and AO/EB kit . RESULTS The best purification technology of total triterpenoid crude extracts from I. obliquus was as follows :AB-8 macroporous adsorption resin was used ;mass concentration of the sample solution was 2.0 mg/mL;sample volume was 140 mL,and the flow rate was 1.0 mL/min;the impurity was removed with 50% ethanol 40 mL, then eluted with 95% ethanol 160 mL,at the elution flow rate of 3.0 mL/min. After purification ,mass concentration of total triterpenoid from I. obliquus increased from 34.36% to 73.39%. The total triterpenoid of I. obliquus could inhibit the proliferation of HeLa cells ,and the 50% inhibitory concentration was 184.20 μg/mL. Compared with control group ,the purified products could significantly inhibit the migratio n and promote the apoptosis of HeLa cells (P<0.05 or P<0.01). CONCLUSIONS The purification technology of total triterpenoids extracts from I. obliquus is successfully optimited . The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.