Synergistic Effect of Methanol Extract of Salvia Miltiorrhiza and Antibiotics against Dental Caries Pathogens

치아우식증유발세균에 대한 단삼 메탄올추출물과 항생제와의 병용효과

  • Jang, Keoun-Ae (Department of Dental Hygiene, MaSan University) ;
  • Kim, Hye-Young (Department of Dental Hygiene, College of Natural Sciences, Dongeui University)
  • Received : 2010.04.26
  • Accepted : 2010.06.21
  • Published : 2010.09.28

Abstract

Salvia miltiorrhiza Bunge (S. miltiorrhiza) is a traditional Korean medicine that is commonly used for the treatment of inflammatory diseases such as edema, arthritis, and hepatitis. The present study investigated the antimicrobial activity of methanol (MeOH) extract of S. miltiorrhiza roots against oral bacteria using broth the microdilution method and the checkerboard and time-kill methods evaluated the synergistic effects of treatment with antibiotics. The MeOH extract was demonstrated as a higher antibacterial activity (MICs, 8 to $64\;{\mu}g/mL$; MBCs, 16 to $64\;{\mu}g/mL$) against all tested oral bacteria. Additionally, the extract was observed to have a synergistic effect with ampicillin or gentamicin. A time-kill study evaluating the effects of the extract indicated that the extract treatment in combination with ampicillin or gentamicin showed rapid bactericidal activity. The results suggest that MeOH extract of S. miltiorrhiza could be employed as a natural antibacterial agent against dental caries.

단삼(Salvia miltiorrhiza Bunge)은 우리나라에서 전통적으로 부종, 관절염, 간염 등과 같은 염증질환의 치료에 사용해왔다. 본 연구에서는 단삼 메탄올 추출물을 이용하여 구강미생물에 대한 항균활성을 확인하였다. 그 결과 메탄올추출물은 실험 되어진 모든 구강미생물에서 강한 항균활성을 나타내었다(MICs, 8 to $64\;{\mu}g/mL$; MBCs, 16 to $64\;{\mu}g/mL$). 추가적으로 ampicillin이나 gentamicin과의 병용투여서 최소억제농도(MIC)와 최소살균농도(MBC)가 감소하는 병용효과를 나타내었다. 더불어 메탄올추출물 단독 사용시 보다 항생제와 병용투여시 빠른 사멸효과를 보였다. 결론적으로 단삼 메탄올추출물이 치아우식을 유발하는 많은 세균들에 대한 항균효과가 뛰어남을 확인 할 수 있었으며, 안정성이 높은 천연 치아우식예방제로서의 개발가능성을 확인하였다.

Keywords

References

  1. Adams, J. D., R. Wang, J. Yang, and E. J. Lien. 2006. Preclinical and clinical examinations of Salvia miltiorrhiza and its tanshinones in ischemic conditions. Chin Med 1: 3. https://doi.org/10.1186/1749-8546-1-3
  2. Cao, J., Y. J. Wei, L. W. Qi, P. Li, Z. M. Qian, H. W. Luo, J. Chen, and J. Zhao. 2008. Determination of fifteen bioactive components in Radix et Rhizoma Salviae Miltiorrhizae by high-performance liquid chromatography with ultraviolet and mass spectrometric detection. Biomed Chromatogr 22: 164-172. https://doi.org/10.1002/bmc.911
  3. Cha, J. D., M. R. Jeong, S. I. Jeong, and K. Y. Lee. 2007. Antibacterial activity of sophoraflavanone G isolated from the roots of Sophora flavescens. J Microbiol Biotechnol 17: 858-864.
  4. Chen, T. H., Y. T. Hsu, C. H. Chen, S. H. Kao, and H. M. Lee. 2007. Tanshinone IIA from Salvia miltiorrhiza induces heme oxygenase-1 expression and inhibits lipopolysaccharide- induced nitric oxide expression in RAW 264.7 cells. Mitochondrion 7: 101-105. https://doi.org/10.1016/j.mito.2006.11.018
  5. Chen, X. G., Y. Li, C. H. Yan, L. N. Li, and R. Han. 2001. Cancer chemopreventive activities of S-3-1, a synthetic derivative of danshinone. J Asian Nat Prod Res 3: 63-75. https://doi.org/10.1080/10286020108042840
  6. Feng, H., H. Xiang, J. Zhang, G. Liu, N. Guo, X. Wang, X. Wu, X. Deng, and L. Yu. 2009. Genome-wide transcriptional profiling of the response of Staphylococcus aureus to cryptotanshinone. J Biomed Biotechnol 23: 1-8.
  7. Hu, P., Q. L. Liang, G. A. Luo, Z. Z. Zhao, and Z. H. Jiang. 2005. Multi-component HPLC fingerprinting of Radix Salviae Miltiorrhizae and its LC-MS-MS identification. Chem Pharm Bull (Tokyo) 53: 677-683. https://doi.org/10.1248/cpb.53.677
  8. Isnansetyo, A. and Y. Kamei. 2003. Pseudoalteromonas phenolica sp. nov., a novel marine bacterium that produces phenolic anti-methicillin-resistant Staphylococcus aureus substances. Int J Syst Evol Microbiol 53: 583-588. https://doi.org/10.1099/ijs.0.02431-0
  9. Jeon, S. J., K. H. Son, Y. S. Kim, Y. H. Choi, and H. P. Kim. 2008. Inhibition of prostaglandin and nitric oxide production in lipopolysaccharide-treated RAW 264.7 cells by tanshinones from the roots of Salvia miltiorrhiza bunge. Arch Pharm Res 31: 758-763. https://doi.org/10.1007/s12272-001-1223-4
  10. Ji, H. S., F. Yu, and J. Yang. 2008. Comparative research on pharmacodynamics of Danshen co-microemulsion on hemorheology in rats with hyperlipidemia. Zhong Yao Cai 31: 566-569.
  11. Kim, J. S., A. S. Narula, and C. Jobin. 2005. Salvia miltiorrhiza water-soluble extract, but not its constituent salvianolic acid B, abrogates LPS-induced NF-kappaB signalling in intestinal epithelial cells. Clin Exp Immunol 141: 288-297. https://doi.org/10.1111/j.1365-2249.2005.02844.x
  12. Lee, J. W., Y. J. Ji, S. O. Lee, and I. S. Lee. 2007. Effect of Saliva miltiorrhiza bunge on antimicrobial activity and resistant gene regulation against methicillin-resistant Staphylococcus aureus (MRSA). J Microbiol 45: 350-357.
  13. Li, Y. I., G. Elmer, and R. C. Leboeuf. 2008. Tanshinone IIA reduces macrophage death induced by hydrogen peroxide by upregulating glutathione peroxidase. Life Sci 83: 557-562. https://doi.org/10.1016/j.lfs.2008.08.003
  14. Liu, A. H., L. Li, M. Xu, Y. H. Lin, H. Z. Guo, and D. A. Guo. 2006. Simultaneous quantification of six major phenolic acids in the roots of Salvia miltiorrhiza and four related traditional Chinese medicinal preparations by HPLC-DAD method. J Pharm Biomed Anal 41: 48-56. https://doi.org/10.1016/j.jpba.2005.10.021
  15. Nie, R., R. Xia, X. Zhong, and Z. Xia. 2007. Salvia miltiorrhiza treatment during early reperfusion reduced postischemic myocardial injury in the rat. Can J Physiol Pharmacol 85: 1012-1019. https://doi.org/10.1139/Y07-092
  16. Ouyang, X., K. Takahashi, K. Komatsu, N. Nakamura, M. Hattori, A. Baba, and J. Azuma. 2001. Protective effect of Salvia miltiorrhiza on angiotensin II-induced hypertrophic responses in neonatal rat cardiac cells. Jpn J Pharmacol 87: 289-296. https://doi.org/10.1254/jjp.87.289
  17. Sekino, S., and P. Ramberg. 2005. The effect of a mouth rinse containing phenolic compounds on plaque formation and developing gingivitis. J Clin Periodontol 32: 1083-1088. https://doi.org/10.1111/j.1600-051X.2005.00793.x
  18. Tang, F., X. Wu, T. Wang, P. Wang, R. Li, H. Zhang, J. Gao, S. Chen, L. Bao, H. Huang, and P. Liu. 2007. Tanshinone II A attenuates atherosclerotic calcification in rat model by inhibition of oxidative stress. Vascul Pharmacol 46: 427-438. https://doi.org/10.1016/j.vph.2007.01.001
  19. Viskelis, P., M. Rubinskiene, I. Jasutiene, A. Sarkinas, R. Daubaras, and L. Cesoniene. 2009. Antibacterial activity of sphagnum acid and other phenolic compounds found in Sphagnum papillosum against food-borne bacteria. J Food Sci 74: C157-C161. https://doi.org/10.1111/j.1750-3841.2009.01066.x
  20. Wu, J. Y., J. Ng, M. Shi, and S. J. Wu. 2007. Enhanced secondary metabolite (tanshinone) production of Salvia miltiorrhiza hairy roots in a novel root-bacteria coculture process. Appl Microbiol Biotechnol 77: 543-550. https://doi.org/10.1007/s00253-007-1192-5
  21. Wu, Z. M., T. Wen, Y. F. Tan, Y. Liu, F. Ren, and H. Wu. 2007. Effects of salvianolic acid a on oxidative stress and liver injury induced by carbon tetrachloride in rats. Basic Clin Pharmacol Toxicol 100: 115-120. https://doi.org/10.1111/j.1742-7835.2007.00020.x
  22. Xu, M., H. Guo, J. Han, S. F. Sun, A. H. Liu, B. R. Wang, X. C. Ma, P. Liu, X. Qiao, et al. 2007. Structural characterization of metabolites of salvianolic acid B from Salvia miltiorrhiza in normal and antibiotic-treated rats by liquid chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 858: 184-198. https://doi.org/10.1016/j.jchromb.2007.08.032
  23. Yin, H. Q., Y. J. Choi, Y. C. Kim, D. H. Sohn, S. Y. Ryu, and B. H. Lee. 2009. Salvia miltiorrhiza Bunge and its active component cryptotanshinone protects primary cultured rat hepatocytes from acute ethanol-induced cytotoxicity and fatty infiltration. Food Chem Toxicol 47: 98-103. https://doi.org/10.1016/j.fct.2008.10.018
  24. Yin, H. Q., Y. S. Kim, Y. J. Choi, Y. C. Kim, D. H. Sohn, S. Y. Ryu, and B. H. Lee. 2008. Effects of tanshinone IIA on the hepatotoxicity and gene expression involved in alcoholic liver disease. Arch Pharm Res 31: 659-665. https://doi.org/10.1007/s12272-001-1209-2
  25. Yu, X. Y., S. G. Lin, Z. W. Zhou, X. Chen, J. Liang, W. Duan, X. Q. Yu, J. Y. Wen, B. Chowbay, et al. 2007. Tanshinone IIB, a primary active constituent from Salvia miltiorrhza, exhibits neuro-protective activity in experimentally stroked rats. Neurosci Lett 417: 261-265. https://doi.org/10.1016/j.neulet.2007.02.079
  26. Yuan, D., Y. N. Pan, W. W. Fu, T. Makino, and Y. Kano. 2005. Quantitative analysis of the marker compounds in Salvia miltiorrihiza root and its phytomedicinal preparations. Chem Pharm Bull (Tokyo) 53: 508-514. https://doi.org/10.1248/cpb.53.508
  27. Zeng, G., H. Xiao, J. Liu, and X. Liang. 2006. Identification of phenolic constituents in Radix Salvia miltiorrhizae by liquid chromatography/electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom 20: 499-506. https://doi.org/10.1002/rcm.2332
  28. Zhao, G. R., H. M. Zhang, T. X. Ye, Z. J. Xiang, Y. J. Yuan, Z. X. Guo, and L. B. Zhao. 2008. Characterization of the radical scavenging and antioxidant activities of danshensu and salvianolic acid B. Food Chem Toxicol 46: 73-81. https://doi.org/10.1016/j.fct.2007.06.034