Study On Active Substance Screening And Resistance Mechanism Of Chinese Gall And Rhizoma Cimicifugae On Oral Pathogenic Bacteria Related To Caries

Caries is a chronic bacterial disease, the bacterium is a necessary condition for the occurrence of dental caries. Since the drugs commonly used anti-caries there are some defects, so many scholars shifted their attention to traditional Chinese medicine which is more secure and have no drug resistant. Our research group found that in clinical work, gall and cohosh is often used in the treatment of oral diseases and the effect is remarkable. Access to relevant Chinese Medicine also found that gall, cohosh have a higher frequency use in the treatment of oral diseases. This study selected the caries bacteria UA159, Actinomyces viscosus, Pg, Ef, Fn, Candida albicans as tested bacteria studied the inhibitory effect of gall, cohosh ethanol aqueous extract to these six kinds of tested bacteria. This study is divided into three parts, namely:gall, Cimicifuga extraction and separation; gall, cohosh antibacterial filter active substance; the influence of reactive monomer screened from gallic on the growth and metabolism of six kinds of oral pathogens. In the first part of the experiment, the compounds extraction from Galla Chinensis and cohosh used heating reflux method with aqueous ethanol. Sequentially extracted compound separated into rough points with organic solvent petroleum ether and ethyl acetate which have large difference in polar. In the second part of the test, including two sets of experiments, i.e. screening antibacterial active substance and separation and purification of the compounds. During the experiment, both sets of experiments alternately. Isolated and purified by means of a series of separation and purification means. Firstly, normal phase silica gel column chromatography was used to get coarse segmentation. Then using reverse phase column chromatography on silica gel, dextran gel column chromatography, high performance liquid chromatography and recrystallization a step by step method of to purify each fraction until having monomer. Finally, three active monomer was acquired and infered the three active monomer as gallic acid, gallic methyl and ethy by post-test analysis of nuclear magnetic resonance, mass spectrometry, etc. Antibacterial experiment used Oxford cup method. In the third part of the experiment, we explore the affect of gallic gallic acid, methyl gallate, and ethyl gallate extracted from gallic on growth,acidogenic and exopolysaccharides to Six oral pathogens. Specific results are as follows:1. gallic acid on UA159, Actinomyces viscosus, Ef, Fn and Candida albicans MIC value are 5mg/ml, MBC values are 10 mg/ml. MIC values and MBC values for Pg are 2.5mg/ml,5mg/ml.Methyl gallate on UA159, Actinomyces viscosus, Ef, Fn and Candida albicans MIC values are 4mg/ml, MBC values are 8 mg/ml. MIC values and MBC values for Pg are 2 mg/ml,4mg/ml. Ethyl gallate on UA159, Actinomyces viscosus, Fn and white Candida MIC values are 2.5mg/ml, MBC values are 5 mg/ml. For Ef and Pg the MIC is 1.25 mg/ml, MBC value of 2.5mg/ml. By comparison, ethyl gallate has strongest inhibitory effect. Compared to other bacteria, gallic acid, gallic acid methyl ester have a stronger inhibitory effect on Pg.2. For the inhibition of oral bacteria producing acid role, gallic acid ethyl ester have a s stronger inhibition of acid production than gallic acid and methyl gallate to Actinomyces, Candida albicans,2mg/ml of ethyl gallate have the equivalent effect with chlorhexidine concentration of 0.25mg/ml. And methyl gallate ester on Porphyromonas gingivalis bacteria cell inhibition of acid production was stronger than gallic acid, 1mg/ml of methyl gallate and ethyl and 0.06mg/ml chlorhexidine solution considerably. For inhibiting bacteria produce extracellular polysaccharide role, gallic acid, methyl gallate ability to suppress white Candida exopolysaccharide weaker than gallic acid ethyl ester. Methyl gallate, ethyl viscous put inhibition actinomycetes produce extracellular polysaccharide stronger than gallic acid. Ethyl gallate on Streptococcus mutans extracellular polysaccharide produced significant inhibitory effect, at the same concentration, the inhibition was stronger than gallic acid and methyl gallate.