Dissertation > Excellent graduate degree dissertation topics show

Experimental Study of Rubrus Suarissimus S.Lee Saponin on Streptococcus Mutans in Vitro

Author: HuangLiWei
Tutor: HeKeXin
School: Guangxi Medical University
Course: Oral medicine
Keywords: Sweet tea saponins Streptococcus mutans Acid production Adhere to Glycosyltransferase Cell external water insoluble polysaccharides
CLC: R151
Type: Master's thesis
Year: 2010
Downloads: 90
Quote: 3
Read: Download Dissertation

Abstract

Purpose by in vitro studies sweet tea extracts - sweet tea glycosides on oral cariogenic bacteria Streptococcus mutans growth, adhesion, and metabolism of plants of containing the natural sweetener - Guangxi sweet tea mechanism of inhibition of Streptococcus mutans, provide experimental basis for the future development and utilization of sweet tea saponins. 1 sweet tea the glycoside solution and broth preparation fold dilution method, the sweet tea the glycosides the preparation of 40g/100ml to 0.125g/100ml solution TTY medium. Of the S.mutans TSB medium activation after 3h inoculated in the MSB a solid medium 37. C under anaerobic culture for 48 hours, typical colonies were picked in 5 ml of sterile saline to prepare a suspension, the density value (Optical Density, OD) of the dimming of the bacterial suspension of 1.0 to calculate the concentration of the bacteria, using bacteria plate count . 2, sweet glycosides on the growth of Streptococcus mutans tube dilution method for the determination of sweet tea saponins collect various in vitro minimum inhibitory concentration (Minimal inhibitory concentration, MIC) size, and centrifuged of S.mutans growth of bacteria, placed in 2 ml of saline , mixing, UV-Vis spectrophotometer OD540 nm value. 3 the, hupehensis glycosides solution of S.mutans acid production according hupehensis glycosides S.mutans MIC measurement result, select the five concentrations below the MIC values, containing 1% sucrose TTY liquid medium dilution, inoculated with equal amounts of variable chain bacterial suspension, 37. C under anaerobic culture for 48 hours, and △ pH values ??before and after culture was determined, and △ pH values ??with the same concentration of xylitol contrast. 4, the influence of the solution of the adhesion of S.mutans hupehensis glycosides taken five concentrations below the MIC, TTY liquid medium containing 1% sucrose hupehensis glucoside solution fold dilution, the solution was mixed with broth hupehensis glycosides 10: a proportional vaccination, all tubes 30 ° horizontal angle anaerobic culture 48h after the test the wall of the bacteria adhered to the elution with NaOH solution, measuring OD540 and calculate the adhesion inhibitory rate, with the viscosity of the same concentration of xylitol attached inhibition rate comparison. 5, the extraction of the glycosyltransferase (Glucosyltransferase, GTF) and water-insoluble following the detection of the extracellular polysaccharide (Insoluble exopolysaccharides) taken MIC concentration of 5, the with TTY liquid medium formulated with different concentrations of hupehensis glycosides solution 3ml added S . mutans broth 0.3ml, After anaerobic culture the bacteria culture was centrifuged, the supernatant was collected, added solid ammonium sulfate, after centrifugation, the precipitate was dissolved in phosphate buffer and dialyzed to obtain the GTF crude enzyme. Coomassie blue method (Bradford method) Determination of the liquid total protein content using Smogyi method determination of the amount of reducing sugars in the enzyme substrate reaction solution. The GTF activity unit (IU) is defined as the standard conditions (37 ℃ for 1h per minute from the amount of enzyme required to release 1μmol reducing sugar sucrose), and calculate the specific activity of the enzyme. Added to the precipitate (cell) of 0.5mol / L NaOH 5ml centrifugation, and the supernatant added 3-fold volume of ethanol, centrifuged, and the precipitate is the reaction of water-insoluble extracellular polysaccharides, using anthrone method Determination of water-insoluble extracellular polysaccharide content. Xylitol with the same concentration of the contrast of S.mutans Enzymes and insoluble extracellular polysaccharides. 1, the experimental tube dilution method, sweet tea saponin concentration in the more medium 20g/100ml growth S.mutans, concentration in the medium below 10g/100ml have S.mutans growth, and with the sweet tea saponins varying chain bacteria concentration decreases, the solution concentration increases. The experimental results determine value of its MlC is 20g/100ml; and sweet tea saponin concentration greater than or equal to 0.313g/100ml, the sweet tea saponins the group S.mutans the OD value overall number is lower than the negative control group (P lt; 0.01) that when n ≥ 0.313 when inhibitory on S.mutans growth, and its inhibition increased with sweet tea saponin concentration increases. Acid production experiments, the pH value of each concentration group compared with before the experiment decreased acid production of xylitol group more so that the pH value decrease. Various sweet tea saponins group before and after the experiment △ pH (pH variation) of the overall mean were differences (P lt; 0.05) 1.25 g/100ml concentration of sweet tea saponins, S. mutans △ pH value is the smallest change in ; of sweet tea glycosides group △ pH value overall mean significantly lower than the xylitol group and negative control group △ pH (P lt; 0.001). 3 With sweet tea saponins concentration increased, the rate of adherence inhibition also will be increased. The the OD value overall concentration of sweet tea saponins group adhesion broth mean significantly below the xylitol group and negative control group (F = 98.02, p lt; 0.01); sweet tea saponin concentration group 5 g/100ml, 2.5 g / 100ml, 1.25 g/100ml outside, among other groups, with a significant difference (p lt; 0.001). 4, total protein, the sweet tea saponins solution in S.mutans of enzyme solution GTF enzyme activity, water insoluble exopolysaccharide of 4.1 with the sweet tea saponins concentration increased, the total protein content of S. mutans enzyme solution is also increasing. Higher than the total protein content of different concentrations of sweet tea saponin group xylitol group and negative control group (P lt; 0.01); xylitol group (P lt; 0.01) is higher than the negative control group; sweet tea saponin concentration group in addition to 0.625 g/100ml, 0.313 g/100ml, the rest of the groups total protein content of the overall mean average with a significant difference (p lt; 0.001). 4.2 With hupehensis glycoside concentration of the solution gradually increased, of S. mutans GTF activity gradually decreased. Negative control group, the amount of reducing sugars, enzyme activity and specific activity were higher than hupehensis glucoside group, xylitol group (p lt; 0.05); enzyme activity of different concentrations of xylitol group was significantly higher than the group of hupehensis glycosides (p LT; 0.001); sweet tea the glucoside group in addition to 5 g/100ml, 2.5 g/100ml 1.25 g/100ml the rest of the group, the amount of reducing sugar, enzyme activity, compared with the vitality of the overall mean difference was significant (p lt; 0.001) . The 4.3 with sweet tea the glycosides solution concentration gradually increased S. mutans generated water insoluble the exopolysaccharide content decreased; negative control group S.mutans synthesis of water-insoluble extracellular polysaccharide ability significantly above the sweet tea saponins and xylitol group (p lt; 0.001); sweet tea saponin concentration group in addition to the rest of the group to 5 g/100ml, 2.5 g/100ml, 1.25 g/100ml with a significant difference (p lt; 0.001). By the Pearson correlation test, sweet glycosides group and xylitol group GTF activity of IS content were positively correlated. Conclusion 1 sweet tea saponins S.mutans growth was significantly inhibited. 2, sweet glycosides can be suppressed S.mutans acid production, and a significant suppression of S.mutans on the surface adhesion effect, and its inhibitory effect was significantly higher than the same concentration of xylitol. 3, sweet glycosides grape glycosyltransferase activity of S.mutans broth has a certain effect, reducing the S.mutans water insoluble extracellular polysaccharide synthesis, significantly inhibited compared with the same concentration of xylitol. 4, in the area of ??dental caries prevention, as a natural plant hupehensis extract, sweet glycosides expected to become a new sweetener.

Related Dissertations

CLC: > Medicine, health > Preventive Medicine,Health > Nutrition, hygiene,food hygiene > Nutrition
© 2012 www.DissertationTopic.Net Mobile