| Objective: Based on the basic theory of Chinese medicine in the treatment of pulmonary tuberculosis,we studied the effects of Coptis chinensis on patients and pathogenic bacteria,and explored the role and mechanism of Coptis chinensis in the treatment of tuberculosis.Methods: 1.The relationship between Coptis chinensis and PTB was investigated by network pharmacology methods.2.The ethanolic extraction of Coptis chinensis and the soxhlet extraction method were used to extract Coptis chinensis with petroleum ether,dichloromethane,ethyl acetate,acetone and anhydrous ethanol,whose polarities ranged from small to large,to detect the effect of the total ethanolic extraction of Coptis chinensis and the different extraction sites on Mycobacterium smegmatis and biofilms.3.The active parts of Coptis chinensis were separated and purified by Sehadex TMLH-20 column separation,to obtain different active fractions,To explore whether different extracted parts of Coptis chinensis have effects on M.smegmatis and biofilms.4.The activity of monomeric components in Coptis chinensis,such as Coptisine,Berberine,Jatrorrhizine,Palmatinehydrochloride,Magnoflorine,Epiberberine and palmatinewas tested to determine which monomeric components have activity to inhibit biofilm formation in M.smegmatis.Results: 1.Possible active ingredients in Phellodendron that exerted pharmacological effects against TB were found to be Berberine,(R)-Canadine,quercetin,Berlambine,Palmatine,Corchoroside A＿qt,Magnograndiolide and Worenine;Among the predicted targets of action RELA,MAPK1,TNF,AKT1,JUN and other target genes may be the main targets of Coptis chinensis in the treatment of PTB.Through the KEGG signaling pathway,in addition to the anti-tuberculosis pathway of action of Coptis chinensis,there is also the AGERAGE signaling pathway acting in cancer pathways,hepatitis B and C,and diabetic complications.2.Coptis chinensis has an inhibitory effect on scale M.smegmatis and its biofilm.The inhibition compounds of M.smegmatis biofilm were mainly concentrated in the anhydrous ethanol site and acetone site,and the acetone site had the best activity in inhibiting M.smegmatis biofilm.The inhibition of biofilm activity was in the following order: acetone >anhydrous ethanol > methylene chloride > petroleum ether > ethyl acetate.3.Then 18 compounds were obtained after activity tracking and separation of the acetone part of Flavobacterium flavum by 60% ethanol through chromatographic column,and their active compounds for biofilm inhibition of M.smegmatis were mainly concentrated in compound 1,compound 5 and compound 6 parts,and compound 1 completely inhibited biofilm growth at a final concentration of 0.003125 mg/ml.4.The effects of the known components of Coptisine,Berberine,Jatrorrhizine,Palmatine hydrochloride,Magnoflorine,Epiberberine and palmatine on biofilm formation showed that the minimum concentrations of Coptisine,Berberine,Jatrorrhizine,Palmatine hydrochloride and palmatine to inhibit biofilm were: 0.01675 mg/ml,0.0125 mg/ml,0.025 mg/ml,0.05 mg/ml,0.0225 mg/ml.Magnoflorine,Epiberberine,did not inhibit biofilm formation at the concentrations tested.Conclusions: 1.The possible active ingredients of Coptis chinensis that exert anti-TB efficacy by supporting righteousness and dispelling evil are Berberine et al.Among the predicted targets of action RELA,MAPK1,TNF,AKT1,JUN and other target genes may be the main targets of Coptischinensis in the treatment of pulmonary tuberculosis.2.The anti-TB efficacy of Coptis chinensis may be due to the inhibition of M.smegmatis and biofilm.3.The inhibitory effect of the mixture of M.smegmatis and biofilm is better than that of some known monomers of Coptis chinensis,indicating the advantages of multi-component and multi-target traditional Chinese medicine therapy.4.The chemical components of Coptis chinensis,such as Berberine,can inhibit both M.smegmatis and biofilm,and have a dual effect,reducing drug treatment time and providing a theoretical basis for clinical treatment. |
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