Screening Of Geranium Strictipes Anticaries Activity Site And The Active Ingredient

Objective: Study effect of Geranium strictipes extract on early artificial root caries and separation compound of Geranium strictipes anticaries activity site and screening of its active constituents to provide a theoretical basis for the prevention and treatment root caries of Geranium strictipes.Methods: (1) Firstly, respectively determine minimum inhibitory concentration (MIC)and minimum bactericidal concentration (MBC) of Geranium strictipes extract on cariogenic bacteria by asp-agar-diluted method and the acid production were determined;(2) Early artificial root caries model was induced by Streptococcus mutans, determining minimum inhibitory concentration (MIC) of Geranium strictipes extract on Streptococcus mutans by asp-agar-diluted method, Using 20.00 mg/LNaF as the positive control group,with purple Sanguisorba officinalis extract treatment of early root surface caries in different concentrations, add fresh broth by 24h, 24h, 48h, 72h three at the time of dissolution of calcium detection and scanning electron microscope observation of the effect of extract of Geranium strictipes remineralization deposition surface of root tissue mineral artificial root caries inhibition of root caries by qualitative analysis of Geranium strictipes; The models of early root caries were constructed by means of cariogenic bacteria, "open window" and put them to 37? for 96 h in demineralization solution to be artificial root caries. six samples were randomly divided into each group. The morphological changes and the changes of mineral content after mineralization were observed by polarized light microscope and confocal laser scanning microscope; (3) Firstly,respectively determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active compounds of Geranium strictipes by different methods on Streptococcus mutans and Streptococcus sanguis and then the active compounds of Geranium strictipes on the acid production_were determined; (4) Using silica gel, Sephadex LH-20 gel, diaion HP-20, MCI gel CHP-20P chromatographic separation technology on the extraction of acetic ether extract of extract of separation and purification, structure analysis on the separation of compounds by magnetic resonance spectroscopy (NMR) method.Results: (1) The extractives of Geranium strictipes can effectively inhibit the growth and acid production of oral bacteria. (2) The minimum bacteriostatic concentration (MIC)of N-butyl alcohol, ethyl acetate and ethanol extractives of Geranium strictipes extracted reflux on Streptococcus mutans is 4.000 g/L, 4.000 g/L, 8.000 g/L, The extractives of Geranium strictipes can effectively extract on the development of root caries in the minimum inhibitory concentration inhibition; The results of polarizing microscopy and confocal laser scanning analysis showed that the extractives of Geranium strictipes on early root surface caries remineralization effect: ethyl acetate extract and n-butanol extract >ethanol extract > water extract of Geranium strictipes,the positive effect of remineralization are ethyl acetate extract and n-butanol extract of 4.5 mg/mL. (3) The 4 active compounds of Geranium strictipes (Gallic acid, Methyl esters of gallic acid,Catechins, Ellagic acid) can affect the growth and metabolism of cariogenic bacteria and effectively inhibit the acid production; (4) First spearate from the n-butanol extract of Geranium strictipes in 4 chemical constituents: daucosterol; leanolic acid 3-O-?-D-ylopyranoside; 3,2'-Epilarixinol;3, 3',4-tri-methylellagic;acid-4'-O-?-D-glucopyranoside.Conclusion: the extractives of Geranium strictipes have inhibitory effects on resident oral bacterial growth and acid production, and can inhibit the root caries demineralization and promote remineralization, thus effectively inhibiting the development of artificial early root surface caries, which is the batter effect of Geranium strictipes n-butanol extract and ethyl acetate extrac, the 4 active compounds of Geranium strictipes(Gallic acid, Methyl esters of gallic acid, Catechins, Ellagic acid) can affect the growth and metabolism of cariogenic bacteria and effectively inhibit the acid production,daucosterol, oleanolic acid 3-O-?-D-ylopyranoside, 3,2'-Epilarixinol and 3,3',4-tri-methylellagicacid-4'-O-?-D-glucopyranoside were spearate from the n-butanol extract of Geranium strictipes extracts.