| Background:Porphyromonas gingivalis is a periodontal tissue opportunistic pathogen, which plays an important role in the occurrence and development of periodontitis. Oral biofilm contains a variety of microbial cells, extracellular polysaccharide, water and cell metabolites. Fromation of biofilm is a dynamic process. With the development of the biofilm, the status and gene expression of bacteria are different. The extracellular polysaccharide constitutes the skeleton of the biofilm. If the expression of polysaccharide is inhibited or destroyed, the stability and integrity of the biofilm will be affected dramatically. In recent years, some studies have shown that berberine may inhibit the growth of P. gingivalis. However, there is no study focusing on the molecular biological mechanism of berberine---- inhibiting the proliferation of bacteria and the synthesis of oral biofilm. Gingival bleeding and infection due to P. gingivalis in the subgingival plaque is the main reason causing periodontitis. With the development of dental implant technique, implantitis caused by P. gingivalis become a new oral disease. Since more and more recent research suggested considering berberine as a new antibiotic, we tried to study on the mechanism of berberine to investigats the feasibility that berberine inhibit P. gingivalis as a new medicine. According to the fact that our long-term goal is to illustrate the molecular mechanism of berberine inhibiting the expression of P. gingivalis:1?Berberine can inhibit the growth of P. gingivalis relation to FtsZ protein.2?Berberine inhibited the expression of P. gingivalis capsular polysaccharide and bacterial biofilm. Objective:(1)Explore the molecular mechanisms of berberine on inhibition the grouth of P. gingivalis.(2)Demonstrate the effect that berberine inhibits the formation of the biofilm of P. gingivalis Method:To test the growth curve of P. gingivalis W83 and P. gingivalis ACTT 33277, the different concentrations of berberine was used for determine the influence of berberine on; Berberine on TSB argar plate tested the concentration of berberine could inhibit P. gingivalis; Electron microscopy and BactlightRed fluorescence staining were performed to detection whether berberine can change P. gingivalis morphologically; to reveal the change of the surface topography of P. gingivalis in different concentration of berberine, Raman spectroscop was used; Real-time fluorescence quantitative PCR was performed to demonstrate the exression of ftsz gene and capsular polysaccharide gene in P. gingivalis with berberine; Biofilm assay tested the expression of biofilm in P. gingivalis under the action of berberine. Result:The over night culture of P. gingivalis was diluted into OD600=0.2, and innoculated in fresh media with 0 mg/ml, 0.01 mg/ml, 0.05 mg/ml, or 0.1 mg/ml of berberine. The growth curve showed that berberine in the concentration of 0.05 mg/ml inhibited the growth of P. gingivalis W83 and ATCC33277. On TSB argar plate with the concentration of berberine as 0 mg/ml, 0.05 mg/ml, 0.1 mg/ml, 0.5 mg/ml, the data showed that there was no P. gingivalis growing around the dot with 0.5 mg/ml of berberine; Electron microscopy and BactlightRed fluorescent staining found that in 0.05 mg/ml berberine 4 hours the shape of P. gingivalis changed comparing to control group(no berberine); Raman spectroscopy results showed that the existence of interaction between berberine and P. gingivalis and alter bacterial traits; Real-tinme PCR results demonstrated the berberine increased the expression of ftsz gene and downregulates capsule gene; The biofilm assay revealed that berberine affect formation of P. gingival biofilm. Conclusion:1?Low concentrations of berberine inhibits the growth of P. gingivalis as the same time change the shape of bacteria, high concentrations may kill P. gingivalis.2?Berberine upregulates the expression of ftsz gene, and inhibit the growth of P. gingivalis.3?Berberine inhibits the formation of P. gingival biofilm and the expression of capsular polysaccharide. |
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