Study On The Screening And Treatment Mechanism Of Traditional Chinese Medicine Monomers Based On Computer Simulation Molecular Docking Technology

Objective: At present,chemotherapy of echinococcosis is mainly based on the benzimidazoles medicine,of which benzimidazoles is one of most effective drug against echicoccosis.However,for its long-term treatment cycle(more than 6 months),the obvious side effects have been observed in human during clinical application.Therefore,it is necessary to find new and effective anti-hydatid drugs.In this study,the computer simulation of molecular docking technology(CSMDT)was used to assess the combination power between β-tubulin and anti-hydatid drugs that had been already licensed for clinical treatment.And on the basis of the above assessement,the feasibility of the CSMDT using for screen the potential anti-hydatid drugs was explored.Furthermore,the CSMDT was used for screening of traditional chinese medicine monomers to select the potential anti-hydatid drugs.Moreover,the anti-hydatid effect of the selected traditional chinese medicin monomers was assessed by in vitro and in vivo experiments.Methods: The creation of theβ-tubulin model of of echinococcus granulosus(E.g)by using Schrodinger 9.3.5 software.Schrodinger 9.3.5 software was used to verify the molecular docking between benzimidazoles and β-tubulin of E.g.and to explore the molecular docking between Chinese medicine monomers and β-tubulin of E.g..The morphological change and mortality rate of E.g was observed after treating with benzimidazoles and the selected Chinese medicine monomers.Ater the treatment of benzimidazoles and the selected Chinese medicine monomers,those were detected respectively that the alkaline phosphatase activity in the protoscolex culture medium by Alkaline Phosphatase Assay Kit.,the protein level of β-tubulin by Western blot,the mRNA expression of β-tublin by qPCR,the cell vability of HepG2 by MTT assay,the cell cycle and cell apoptosis of HepG2 cells by flow cytometry,the protein and mRNA levels of apoptosis-related proteins in HepG2 cells by Western bloting and Real Time PCR,the cell microtubules formation in HepG2 cells by cytoimmunofluorescence.Results:1.Schrodinger 9.3.5 molecular docking technology demostrated that levorotation albendazole sulfoxide(L-ASOX),dextrorotation(D-ASOX),febantel,nocodazole,mebendazole,benzimidazole and thiabendazole have high combination with β-tubulin in echinococcus granulosa.2.Seven Chinese traditional monomers which have high combination with β-tubulin had been selected by using AutoDock simulated molecular docking technology from Chinese medicine monomers library,including Chinese medicine monomer,PFM(Artemisia myriantha class),NIP(Isoflavone cinnamon class),TPC(TPC(sesquiterpenoid derivative)ASE(anhui shellfish class),ALT(anthocyanin class),DTA(Rhizophora mucronata class)and CPI(Withania somnifera lactide class).3.L-ASOX,D-ASOX,febantel,nocodazole,mebendazole,benzimidazole and thiabendazole exhibited anti-hydatid effect in vitro.After treating with such medicines,the alkaline phosphatase activity was enhanced in protoscolex culture medium.The protein level of β-tubulin was increased and mRNA level was decreased by treating with such medicines.L-ASOX and D-ASOX have the best anti-hydatid effect among such medicines.4.Chinese medicine monomer,PFM,NIP,TPC,ASE,ALT,DTA and CPI exhibiting anti-hydatid effect in vitro.After treating with such Chinese medicin e monomer,the alkaline phosphatase activity was enhanced in protoscolex cultu re medium.The protein level of β-tubulin was increased and mRNA level was decreased by treating with such Chinese medicine monomer.PFM and NIP ha ve the best anti-hydatid effect among such Chinese medicine monomer,5.ASOX,L-ASOX and D-ASOX could inhibit cell vability,promoting cell apoptosis,up-regulating Caspase9,Cyt C and Bax expression,down-regulating Bcl-2 expression,and damaging cell microtubules formation in HepG2 cells.6.PFM and NIP could inhibit cell vability,promoting cell apoptosis,up-regulating Caspase9,Cyt C and Bax expression,down-regulating Bcl-2 expression,and damaging cell microtubules formation in HepG2 cells,and cell cytotoxicity of PFM and NIP was more weaker than ASOX.Conclusion:1.In vitro biology experiment results show that computer simulation molecular docking technology is basically consistent with the effect of the target binding energy of the drug and the hydatid beta-tubulin target.2.The computer simulation molecular docking technology can be applied to the screening of anti-insect drugs,which can be applied to the screening of traditional Chinese medicine monomers.3.The traditional Chinese medicine monomers screened by biological validation and computer simulation molecular docking technology had the activity of antihydatid.4.In the western medicine groups,the albendazole dextrazole sulfone on the effect of anti-hydatid was the most obvious,and in the Chinese medicine monomer groups,PFM and NIP were the most obvious.5.The microtubulin polymerization can be inhibited by the biological validation of the drugs screened by beta-tubulin as the target protein.6.PFM and NIP with better efficacy In vitro biological experiments were less cell toxic than recommended drugs.