Study On Anti-lung Cancer Mechanism Of Xianghai Wild Goose Blood Polypeptide Based On Network Pharmacology And Molecular Docking Technology

Xianghai wild goose is a hybrid of Anser cygnoides and Semen anserinum.In this paper,the polypeptide of Xianghai wild goose blood was extracted,isolated,purified and analyzed.The mechanism of its anti-lung cancer action was analyzed based on network pharmacology and molecular docking technology.Its activity against lung cancer cells was investigated.The research contents are as follows:Using fresh Xianghai wild goose blood as raw material,the degree of hydrolysis was used as index to screen tool enzymes.The single factor experiment was used to determine the enzymatic hydrolysis process of blood polypeptide of Xianghai wild goose.Response surface methodology was used to optimize the extraction process of polypeptides from goose blood.The amino acid composition was analyzed by amino acid analyzer.Sephadex G-25 gel chromatography was used to purify and isolate polypeptides.The results showed that the optimal compound protease was used as tool enzyme.The optimal hydrolysis conditions were as follows:ultrasonic power 540 W,ultrasonic frequency 20 KHz,enzyme dosage 4000 U/g,ultrasonic time 4.5 h,under which the degree of hydrolysis was 28.77%.17 kinds of amino acids were detected,8of which were essential amino acids.XGP was purified by Sephadex G-25 gel chromatography.The composition of XGP was analyzed by UPLC-Q-TOF-MS/MS.The active components,potential targets and molecular mechanism of blood polypeptides in Xianghai wild goose were predicted by network pharmacology method.Including drug similarity assessment,oral bioavailability prediction,protein-protein interaction（PPI）network construction and analysis,gene ontology（GO）terminology and reaction group pathway annotation;Based on network pharmacology,an interaction network of"component-target-pathway"was constructed.Molecular docking simulation was used to further explain the active site and binding degree of ligand and explain its possible mechanism of action.The results show that the components of the chemical constituents are as follows:H-Asn-Asp-Asp-Met-OH,Thr-Thr-Asn-Tyr-Thr-Asp,Ala-Trp-Met-Asp-Phe-Val;A total of 258 related targets were screened by network pharmacology,46 core targets were obtained by PPI screening,such as AKT1,IL1B,SRC,STAT3,MMP9 and CCND1.GO enrichment analysis of 64 items,mainly regulating lung cancer response,protein phosphorylation,redox process and other biological functions,affecting cell proliferation,apoptosis,redox,etc.KEGG analysis enriched 56 signal pathways,mainly including PI3K-ATK,RAS,Hepatitis B and other signal pathways.XGP acts synergistically on lung cancer networks through multiple components,multiple targets and multiple pathways.The P1,P2 and P3 blood peptides had good binding ability to the core target AKT1,SRC,MMP9.It is proved that Xianghai wild goose blood polypeptide has potential inhibitory effect on lung cancer.CCK-8 method was used to detect the inhibitory effect of different concentrations of XGP on A549 cell proliferation,and to detect the intracellular Ca²⁺release,cell oxidative damage and ATP content of different concentrations of XGP.Lung cancer A549 cells were induced by XGP,and the expression levels of key pathway target proteins were detected by q PCR and Western blot.The results showed that XGP inhibited the proliferation of A549 cells in a concentration and time dependent manner.XGP may cause Ca²⁺overload,oxidative damage,inhibition of ATP synthesis and interference of energy metabolism to apoptosis through A549 cells.q PCR results showed that XGP could inhibit the expression level of core target AKT1.Western blot results showed that XGP inhibited lung cancer cell A549 by decreasing AKT1 m RNA expression level.To further verify the inhibitory effect of Xianghai wild goose blood polypeptide on lung cancer.Xianghai wild goose blood polypeptide has broad application prospect in anti-lung cancer,which provides reference basis and theoretical basis for drug development and application of Xianghai wild goose blood in the future.