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Inhibitory Activity of Flower Extracts from Salvia deserta Schang on Streptococcus mutans
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摘要:
目的 研究新疆鼠尾草花提取物（Salvia deserta Schang flower extracts, SFE）对变异链球菌（Streptococcus mutans, S. mutans）的体外抑菌活性。 方法 通过琼脂打孔法和微量稀释法测定SFE对S. mutans浮游菌的抑菌作用及对其生长过程的影响；结晶紫染色法和MTT还原试验测定SFE对S. mutans生物膜的影响；蒽酮-硫酸法测定SFE对细菌生物膜产胞外多糖（exopolysaccharides, EPS）量的影响；乳酸脱氢酶（lactate dehydrogenase, LDH）比色法测定S. mutans的胞内LDH活性；采用pH计测定SFE对S. mutans产酸能力的影响。 结果 SFE对S. mutans的最小抑菌浓度（minimum inhibitory concentration, MIC）为14 μg/μL，1/8 MIC到MIC浓度的SFE在30 h内均能抑制S. mutans的生长速度，且与对照组相比能显著抑制其LDH活性（P<0.0001），4 MIC到1/4 MIC浓度的SFE对S. mutans终产酸量有显著抑制作用（P<0.001），并且能有效抑制S. mutans生物膜的形成，同时能使生物膜产生的EPS量明显减少（P<0.01）。 结论 SFE能有效抑制S. mutans及其生物膜活性，初步探讨其抑制机制为：通过减少细菌EPS的产量来干扰微生物的粘附和聚集，从而抑制细菌生物膜的形成；通过降低细菌LDH活性来干扰S. mutans的糖酵解，从而抑制S. mutans产酸。 Abstract:Objective To examine the in vitro inhibitory effect of flower extracts from Salvia deserta Schang (SFE) on Streptococcu smutans (S. mutans). Methods The inhibitory effect of SFE on planktonic S. mutans and the effect of SFE on the growth process of planktonic S. mutans were determined by the agar drilling method and the microdilution method. Crystal violet staining and MTT reduction assay were conducted to determine the effect of SFE on S. mutans biofilm formation. The effect of SFE on the production of exopolysaccharides (EPS) in S. mutans biofilm was determined by anthrone-sulfuric acid method. The intracellular lactate dehydrogenase (LDH) activity in S. mutans was determined by LDH colorimetric assay. The effects of SFE on the acid-producing capacity of S. mutans was determined by pH meter. Results The minimum inhibitory concentration (MIC) of SFE against S. mutans was 14 μg/μL. SFE of the the concentration between 1/8 MIC and MIC could inhibit the growth rate of S. mutans within 30 h and it could significantly inhibit the LDH activity compared with the control group (P<0.0001). SFE of the concentration between 4 MIC and 1/4 MIC had an inhibitory effect on the acid production of S. mutans (P<0.001). Moreover, it could effectively restrain the formation of S. mutans biofilm and significantly reduce the amount of EPS produced by biofilm (P<0.01). Conclusion SFE can effectively inhibit the activity of S. mutans and its biofilm. The mechanism of inhibiting S. mutans by SFE was preliminarily discussed as follows, it interferes with microbial adhesion and aggregation by reducing the production of bacterial EPS, thus inhibiting the formation of bacterial biofilms. In addition, it interferes with glycolysis of S. mutans by reducing the LDH activity of bacteria, thus inhibiting the acid production of S. mutans. -
Key words:
- Salvia deserta Schang flower /
- Streptococcus mutans /
- Biofilm /
- Lactate dehydrogenase
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图 1 新疆鼠尾草花抑菌活性成分化学结构图
Figure 1. Chemical structure diagram of antibacterial active components in SFE
图 2 S. mutans在不同SFE浓度下的生长曲线
Figure 2. Growth curves of S. mutans under different concentrations of SFE
n=3; * P<0.05, **** P<0.0001, vs. control at 30 h.
图 3 SFE对S. mutans生物膜的影响
Figure 3. Effect of SFE on the biofilm fromation of S. mutans
A: Results of crystal violet assay; B: MTT assay results; C: the effect of SFE treatment on bacterial biofilms was observed by LSCM (SYTO9: green fluorescence, which was used for both the dead and live bacteria; PI: red fluorescence, which was used for dead bacterial cells). n=6; ** P<0.01, **** P<0.0001, vs. control group.
图 4 SFE对S. mutans产EPS、产酸和LDH活性的影响
Figure 4. Effects of SFE on EPS and acid production of S. mutans and its LDH activity
A: Effect of SFE on EPS production of S. mutans; B: effect of SFE on LDH activity of S. mutans; C: effect of SFE on acid production of S. mutans. n=3; ** P<0.01, *** P<0.001, **** P<0.0001, vs. control group.
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[1] HU P, LV B B, YANG K X, et al. Discovery of myricetin as an inhibitor against Streptococcus mutans and an anti-adhesion approach to biofilm formation. Int J Med Microbiol,2021,311: 151512. doi: 10.1016/j.ijmm.2021.151512 [2] ZHANG Y C, ZHU Y, ZUO Y P, et al. Efects of Rhein-8-O-β-D-glucopyranoside on the bioflm formation of streptococcus mutans. Curr Microbiol,2021,78: 323–328. doi: 10.1007/s00284-020-02248-0 [3] DIMOU I, DRITSAS S, AGGELOPOULOU P, et al. A development of a herbal mouthwash containing a mixture of essential oils and plant extracts and in vitro testing of its antimicrobial efficiency against the planktonic and biofilm-enclosed cariogenic bacterium Streptococcus mutans. Biofouling,2021,37(4): 397–409. doi: 10.1080/08927014.2021.1924693 [4] 陈东茹, 林焕彩. 变异链球菌致龋机制研究新进展. 四川大学学报(医学版),2022,53(2): 208–213. doi: 10.12182/20220360508 [5] LYU X, LI C, ZHANG J, et al. A novel small molecule, LCG-N25, inhibits oral streptococcal biofilm. Front microbiol,2021,12: 654692. doi: 10.3389/fmicb.2021.654692 [6] PHILIP N, BANDARA H, LEISHMAN S J, et al. Inhibitory effects of fruit berry extracts on Streptococcus mutans biofilms. Eur J Oral Sci,2019,127: 122–129. doi: 10.1111/eos.12602 [7] 黄晓晶, 江山, 蔡志宇, 等. 生物膜状态变异链球菌临床株合成胞外多糖能力的研究. 重庆医科大学学报,2011,36(3): 281–285. doi: 10.13406/j.cnki.cyxb.2011.03.036 [8] ZAYED S M, ABOULWAFA M, HASHEM A M, et al. Bioflm formation by Streptococcus mutans and its inhibition by green tea extracts. AMB Expr,2021,11: 73. doi: 10.1186/s13568-021-01232-6 [9] BESINIS A, DEPERALTA T, HANDY R D. The antibacterial effects of silver, titanium dioxide and silica dioxide nanoparticles compared to the dental disinfectant chlorhexidine on Streptococcus mutans using a suite of bioassays. Nanotoxicology,2014,8(1): 1–16. doi: 10.3109/17435390.2012.742935 [10] 任春晖, 王晓梅, 王新玲, 等. 新疆鼠尾草根中酚酸类化学成分的纯化工艺研究. 西北药学杂志,2021,36(4): 527–532. doi: 10.3969/j.issn.1004-2407.2021.04.002 [11] 王晓梅, 任春晖, 王新玲, 等. 新疆鼠尾草酚酸类成分对HK-2细胞氧化损伤的保护作用及机制研究. 中国药房,2022,33(19): 2348–2353. doi: 10.6039/j.issn.1001-0408.2022.19.09 [12] ZHUSSPOVA A, ZHUMALIYEVA G, OGAY V, et al. Immunomodulatory effects of plant extracts from Salvia deserta Schang and Salvia sclarea L. Plants,2022,11(20): 2690. doi: 10.3390/plants11202690 [13] 王新玲, 王晓梅, 胡君萍, 等. 新疆鼠尾草不同部位不同极性提取物抗氧化活性. 中国实验方剂学杂志,2017,23(23): 62–65. doi: 10.13422/j.cnki.syfjx.2017230062 [14] WANG L W, WU W X, TIAN S G. Qualitative and quantitative analyses of four active components in different organs of Salvia deserta Schang by high-performance thin-layer chromatography. J Chromatogr Sci,2023,61(3): 225–233. doi: 10.1093/chromsci/bmac014 [15] JEONG M J, LIM D S, JEONG S J, et al. Anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid in lipopolysaccharide-stimulated MC3T3-E1 osteoblastic cells on a titanium surface. J Dent Sci,2020,20: 221–229. doi: 10.17135/jdhs.2020.20.4.221 [16] ALMEIDA A A P, NAGHETINI C C, SANTOS V R, et al. Influence of natural coffee compounds, coffee extracts and increased levels of caffeine on the inhibition of Streptococcus mutans. Food Res Int,2012,49: 459–461. doi: 10.1016/j.foodres.2012.07.026 [17] LIU Y C, HUANG Y X, FAN C, et al. Ursolic acid targets glucosyltransferase and inhibits its activity to prevent Streptococcus mutans biofilm formation. Front microbiol,2021,12: 743305. doi: 10.3389/fmicb.2021.743305 [18] 阿孜古丽·阿里木, 王启文, 刘阳, 等. 高效液相色谱结合Box-Behnken响应面法优选新疆鼠尾草根萜类成分提取纯化工艺. 中国医院药学杂志,2023,43(8): 855–862. doi: 10.13286/j.1001-5213.2023.08.05 [19] 杨艾华, 宋姗姗, 王微微. 火炭母提取物对金黄色葡萄球菌的抑菌活性及稳定性研究. 食品与机械,2021,37(11): 148–152. doi: 10.13652/j.issn.1003-5788.2021.11.026 [20] 吴菊, 王玲, 刘兴容. 黄芩苷对变异链球菌UA159体外的抑制作用. 口腔疾病防治,2021,29(7): 462–467. doi: 10.12016/j.issn.2096-1456.2021.07.005 [21] KIM M A, KIM J H, NAM O H. Tea extracts differentially inhibit Streptococcus mutans and Streptococcus sobrinus biofilm colonization depending on the steeping temperature. Biofouling,2020,36(3): 256–265. doi: 10.1080/08927014.2020.1755429 [22] PRIYA A, SELVARAJ A, DIVYA D, et al. In vitro and in vivo anti-infective potential of thymol against early childhood caries causing dual species Candida albicans and Streptococcus mutans. Front Pharmacol,2021,12: 760768. doi: 10.3389/fphar.2021.760768 [23] ELANGO A V, VASUDEVAN S, SHANMUGAM K, et al. Exploring the anti-caries properties of baicalin against Streptococcus mutans: an in vitro study. Biofouling,2021,37(3): 267–275. doi: 10.1080/08927014.2021.1897789 [24] TAKAHASHI N, NYVAD B. The role of bacteria in the caries process: ecological perspectives. J Dent Res,2011,90: 294–303. doi: 10.1177/0022034510379602 [25] 范骏. 翻译后修饰与肿瘤代谢重编程. 中山大学学报 (医学科学版),2022,43(2): 161–172. doi: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2022.0201 [26] 孙艳伟, 刘雅丽, 赵晓雪, 等. 柠檬精油对变异链球菌产酸及乳酸脱氢酶活性影响的实验研究. 口腔医学研究,2018,34(1): 27–31. doi: 10.13701/j.cnki.kqyxyj.2018.01.007 -
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