大黄素-8-O-β-D-吡喃葡萄糖苷对牙龈卟啉单胞菌唾液酸酶活性及其毒力基因表达影响研究
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摘要
目的探讨大黄素-8-O-β-D-吡喃葡萄糖苷对牙龈卟啉单胞菌(P. gingivalis)唾液酸酶活性及其毒力基因表达的影响。方法使用不同质量浓度的大黄素-8-O-β-D-吡喃葡萄糖苷(0.2、0.5、2、5、10 mg/mL)处理P. gingivalis W83(实验组),用未加药物的P. gingivalis W83作对照(对照组),采用荧光法检测大黄素-8-O-β-D-吡喃葡萄糖苷对P. gingivalis唾液酸酶活性的作用。5 mg/mL大黄素-8-O-β-D-吡喃葡萄糖苷作用于P. gingivalis W83,Real-time PCR法检测毒力基因fimA、fimR、fimS、kgp、rgpA和rgpB的表达情况。结果大黄素-8-O-β-D-吡喃葡萄糖苷对P. gingivalis唾液酸酶活性产生了抑制作用,当其质量浓度为0.2、0.5、2、5、10 mg/mL时,对唾液酸酶活性的抑制率分别为11.4%、32.23%、40.21%、73.54%、84.31%。与对照组比较,实验组(5 mg/mL大黄素-8-O-β-D-吡喃葡萄糖苷处理)的fimA、fimR、fimS、kgp、rgpA和rgpB基因表达均下降,差异均有统计学意义(均P <0.05)。结论大黄素-8-O-β-D-吡喃葡萄糖苷可有效抑制P. gingivalis唾液酸酶活性,其抑制作用会降低细菌毒力基因表达,有望成为预防及治疗牙周炎的新型药物。
Objective To detect the inhibitory effect of emodin-8-O-β-D-glucopyranoside on sialidase activity and virulence genes expression of Porphyromonas gingivalis(P. gingivalis). Methods Different concentrations of emodin-8-O-β-D-glucopyranoside(0.2,0.5,2,5,10 mg/mL)were added in P. gingivalis W83 cultural medium(experiment groups). The control group consisted of the untreated P. gingivalis W83. The inhibitory effect of emodin-8-O-β-D-glucopyranoside on P. gingivalis sialidase was verified by using fluorometric assay. At a concentration of emodin-8-O-β-D-glucopyranoside(5 mg/mL),the effect on virulence genes expression in P. gingivalis(fimA,fimR,fimS,kgp,rgpA and rgpB)was determined by Real-time PCR. Results When the concentrations were 0.2,0.5,2,5 and 10 mg/mL,the inhibition rates of sialidase activity were 11.4%、32.23%、40.21%、73.54% and 84.31%,respectively. Compared with control group,the expressions of fimA,fimR,fimS,kgp,rgpA and rgpB in experiment groups were decreased significantly(P < 0.05). Conclusion Emodin-8-O-β-D-glucopyranoside exhibits effective inhibition on sialidase activities in P.gingivalis,and decreases the expression of gene virulence,which may become a new drug in the prevention and treatment of periodontitis.
Objective To detect the inhibitory effect of emodin-8-O-β-D-glucopyranoside on sialidase activity and virulence genes expression of Porphyromonas gingivalis(P. gingivalis). Methods Different concentrations of emodin-8-O-β-D-glucopyranoside(0.2,0.5,2,5,10 mg/mL)were added in P. gingivalis W83 cultural medium(experiment groups). The control group consisted of the untreated P. gingivalis W83. The inhibitory effect of emodin-8-O-β-D-glucopyranoside on P. gingivalis sialidase was verified by using fluorometric assay. At a concentration of emodin-8-O-β-D-glucopyranoside(5 mg/mL),the effect on virulence genes expression in P. gingivalis(fimA,fimR,fimS,kgp,rgpA and rgpB)was determined by Real-time PCR. Results When the concentrations were 0.2,0.5,2,5 and 10 mg/mL,the inhibition rates of sialidase activity were 11.4%、32.23%、40.21%、73.54% and 84.31%,respectively. Compared with control group,the expressions of fimA,fimR,fimS,kgp,rgpA and rgpB in experiment groups were decreased significantly(P < 0.05). Conclusion Emodin-8-O-β-D-glucopyranoside exhibits effective inhibition on sialidase activities in P.gingivalis,and decreases the expression of gene virulence,which may become a new drug in the prevention and treatment of periodontitis.
引文
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[13] Yoshimura F,Murakami Y,Nishikawa K,et al. Surface components of Porphyromonas gingivalis[J]. J Periodontal Res,2009,44(1):1-12.
[14] Nishikawa K,Yoshimura F,Duncan MJ. A regulation cascade controls expression of Porphyromonas gingivalis fimbriae via the FimR response regulator[J]. Mol Microbiol,2004,54(2):546-560.
[15] Wu J,Lin X,Xie H. Porphyromonas gingivalis short fimbriae are regulated by a FimS/FimR two-component system[J].FEMS Microbiol Lett,2007,271(2):214-221.
[16] Lo AW,Seers CA,Boyce JD,et al. Comparative transcriptomic analysis of Porphyromonas gingivalis biofilm and planktonic cells[J]. BMC Microbiol,2009,9:18.
[17] Potempa J,Pike R,Travis J. Titration and mapping of the active site of cysteine proteinases from Porphyromonas gingivalis(gingipains)using peptidyl chloromethanes[J]. Biol Chem,1997,378(3-4):223-230.
[18] Nakayama K,Yoshimura F,Kadowaki T,et al. Involvement of arginine-specific cysteine proteinase(Arg-gingipain)in fimbriation of Porphyromonas gingivalis[J]. J Bacteriol,1996,178(10):2818-2824.
[19] Hou Y,Yu H,Liu X,et al. Gingipain of Porphyromonas gingivalis manipulates M1 macrophage polarization through C5a pathway[J]. In Vitro Cell Dev Biol Anim,2017,53(7):593-603.
[2] Lewis AL,Lewis WG. Host sialoglycans and bacterial sialidases:a mucosal perspective[J]. Cell Microbiol,2012,14(8):1174-1182.
[3] Varki A. Glycan-based interactions involving vertebrate sialicacid-recognizing proteins[J]. Nature,2007,446(7139):1023-1029.
[4] Li C,Kurniyati,Hu B,et al. Abrogation of neuraminidase reduces biofilm formation,capsule biosynthesis,and virulence of Porphyromonas gingivalis[J]. Infect Immun,2012,80(1):3-13.
[5] Xu X,Tong T,Yang X,et al. Differences in survival,virulence and biofilm formation between sialidase-deficient and W83 wildtype Porphyromonas gingivalis strains under stressful environmental conditions[J]. BMC Microbiol,2017,17(1):178.
[6] Aarnio K,Siegerink B,Pirinen J,et al. Cardiovascular events after ischemic stroke in young adults:A prospective follow-up study[J]. Neurology,2016,86(20):1872-1879.
[7] Uddin Z,Song YH,Curtis-Long MJ,et al. Potent bacterial neuraminidase inhibitors,anthraquinone glucosides from Polygonum cuspidatum and their inhibitory mechanism[J]. J Ethnopharmacol,2016,193:283-292.
[8] James CE,Hasegawa Y,Park Y,et al. LuxS involvement in the regulation of genes coding for hemin and iron acquisition systems in Porphyromonas gingivalis[J]. Infect Immun,2006,74(7):3834-3844.
[9] Hiratsuka K,Hayakawa M,Kiyama-Kishikawa M,et al. Role of the hemin-binding protein 35(HBP35)of Porphyromonas gingivalis in coaggregation[J]. Microb Pathog,2008,44(4):320-328.
[10]佟彤,李琛,徐晓宇,等.不同培养条件下唾液酸酶基因缺失对牙龈卟啉单胞菌生物膜形成的影响研究[J].中国实用口腔科杂志,2017,10(7):436-440.
[11] Aruni W,Vanterpool E,Osbourne D,et al. Sialidase and sialoglycoproteases can modulate virulence in Porphyromonas gingivalis[J]. Infect Immun,2011,79(7):2779-2791.
[12] Amano A. Bacterial adhesins to host components in periodontitis[J]. Periodontol 2000,2010,52(1):12-37.
[13] Yoshimura F,Murakami Y,Nishikawa K,et al. Surface components of Porphyromonas gingivalis[J]. J Periodontal Res,2009,44(1):1-12.
[14] Nishikawa K,Yoshimura F,Duncan MJ. A regulation cascade controls expression of Porphyromonas gingivalis fimbriae via the FimR response regulator[J]. Mol Microbiol,2004,54(2):546-560.
[15] Wu J,Lin X,Xie H. Porphyromonas gingivalis short fimbriae are regulated by a FimS/FimR two-component system[J].FEMS Microbiol Lett,2007,271(2):214-221.
[16] Lo AW,Seers CA,Boyce JD,et al. Comparative transcriptomic analysis of Porphyromonas gingivalis biofilm and planktonic cells[J]. BMC Microbiol,2009,9:18.
[17] Potempa J,Pike R,Travis J. Titration and mapping of the active site of cysteine proteinases from Porphyromonas gingivalis(gingipains)using peptidyl chloromethanes[J]. Biol Chem,1997,378(3-4):223-230.
[18] Nakayama K,Yoshimura F,Kadowaki T,et al. Involvement of arginine-specific cysteine proteinase(Arg-gingipain)in fimbriation of Porphyromonas gingivalis[J]. J Bacteriol,1996,178(10):2818-2824.
[19] Hou Y,Yu H,Liu X,et al. Gingipain of Porphyromonas gingivalis manipulates M1 macrophage polarization through C5a pathway[J]. In Vitro Cell Dev Biol Anim,2017,53(7):593-603.