SC对敏感菌种在口腔内黏附能力的影响
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摘要
目的探讨唾液中分泌片SC对敏感菌黏附口腔上皮细胞能力的影响及机制。方法将敏感菌S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176与来源于60例健康志愿者的口腔上皮细胞进行混合培养,采用革兰阳性染色分别观察加入及未加入唾液中SC时敏感菌的黏附能力,采用荧光定量RT-PCR法分别测定加入及未加入唾液中SC时上述3种敏感菌中ALS2与ALS3 mRNA的表达情况。结果在不含SC的唾液上清液+敏感菌(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176)+口腔上皮细胞体系中,S. sanguis ATCC 10556黏附率最高,三者比较差异有统计学意义(P <0. 05);在含SC的唾液上清液+敏感菌(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176)+口腔上皮细胞体系中,S. sanguis ATCC 10556黏附率最低,三者比较差异有统计学意义(P <0. 05);且含SC体系中,3种敏感菌的黏附率均明显低于不含SC体系中3种敏感菌的黏附率(P均<0. 05)。RTPCR检测结果显示,在不含SC的唾液上清液+敏感菌(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176)+口腔上皮细胞体系中,不含SC唾液上清液+S. sanguis ATCC 10556+口腔上皮细胞体系中ALS2及ALS3 mRNA表达水平最高,三者比较差异有统计学意义(P <0. 05);在含SC的唾液上清液+敏感菌(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176)+口腔上皮细胞体系中,含SC唾液上清液+S. sanguis ATCC 10556+口腔上皮细胞体系中ALS2及ALS3 mRNA表达水平最低,三者比较差异有统计学意义(P <0. 05);且含SC体系中3种敏感菌的ALS2与ALS3 mRNA表达水平均明显低于不含SC体系中3种敏感菌的表达水平(P均<0. 05)。结论不同敏感菌的口腔上皮细胞黏附能力存在差异,菌株S. sanguis ATCC 10556黏附能力强于S. mutans UA159和S. sobrinus OMZ-176。唾液SC能够抑制敏感菌对口腔上皮细胞的黏附,且对菌株S. sanguis ATCC 10556抑制能力更强,可能与下调ALS2和ALS3基因表达相关。
Objective It is to investigate the effect of salivary secretory tablet SC on the adhesive ability of susceptible strains to oral epithelial cells and its mechanism. Methods Susceptible strains S. sanguis ATCC 10556,S. mutans UA159 and S. sobrinus OMZ-176 were cultured by mixed with oral epithelial cells from 60 healthy volunteer S. The adhesive ability of susceptible strains was observed by Gram positive staining before and after SC was added to saliva. The expression of ALS2 and ALS3 mRNA in the three susceptible strains was detected by fluorescence quantitative RT-PCR. Results In the salivary supernatant without SC + sensitive bacteria(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176) +oral epithelial cell system,the adhesion rate of S. sanguis ATCC 10556 was the highest,there was significant difference among the three groups(P < 0. 05). In the salivary supernatant containing SC + susceptible strains(S. sanguis ATCC10556,S. mutans UA159,S. sobrinus OMZ-176) + oral epithelial cell system,the adhesion rate of S. sanguis ATCC10556 was the lowest,there was significant difference among the three groups(P < 0. 05). The adhesion rates of the three susceptible strains in the system containing SC were significantly lower than those in the system without SC(P < 0. 05).The results of RT-PCR showed: In the salivary supernatant without SC + Susceptible strains + oral epithelial cell system,that the expression levels of ALS2 and ALS3 mRNA were highest in salivary supernatant without SC + S. sanguis ATCC10556 + oral epithelial cells system,there was significant difference among the three groups(P < 0. 05); In the salivary supernatant containing SC + Susceptible strains + oral epithelial cell system,the expression levels of ALS2 and ALS3 mRNA in salivary supernatant containing SC + S. sanguis ATCC 10556 + oral epithelial cells were the lowest,there was significant difference among the three group(P < 0. 05). Moreover,the expression levels of ALS2 and ALS3 mRNA of three Susceptible strains in containing SC system were significantly lower than those in without SC system(all P < 0. 05).Conclusion the adhesive ability of oral cavity epithelial cells of different susceptible strains is different. The adhesive ability of strain S. sanguis ATCC 10556 was stronger than that of S. mutans UA159 and S. sobrinus OMZ-176. Salivary SC can inhibit the adhesion of susceptible strains to oral epithelial cells,and the strain S. sanguis ATCC 10556 has stronger inhibition ability,it may be related to down regulation of ALS2 and ALS3 gene expression.
Objective It is to investigate the effect of salivary secretory tablet SC on the adhesive ability of susceptible strains to oral epithelial cells and its mechanism. Methods Susceptible strains S. sanguis ATCC 10556,S. mutans UA159 and S. sobrinus OMZ-176 were cultured by mixed with oral epithelial cells from 60 healthy volunteer S. The adhesive ability of susceptible strains was observed by Gram positive staining before and after SC was added to saliva. The expression of ALS2 and ALS3 mRNA in the three susceptible strains was detected by fluorescence quantitative RT-PCR. Results In the salivary supernatant without SC + sensitive bacteria(S. sanguis ATCC 10556,S. mutans UA159,S. sobrinus OMZ-176) +oral epithelial cell system,the adhesion rate of S. sanguis ATCC 10556 was the highest,there was significant difference among the three groups(P < 0. 05). In the salivary supernatant containing SC + susceptible strains(S. sanguis ATCC10556,S. mutans UA159,S. sobrinus OMZ-176) + oral epithelial cell system,the adhesion rate of S. sanguis ATCC10556 was the lowest,there was significant difference among the three groups(P < 0. 05). The adhesion rates of the three susceptible strains in the system containing SC were significantly lower than those in the system without SC(P < 0. 05).The results of RT-PCR showed: In the salivary supernatant without SC + Susceptible strains + oral epithelial cell system,that the expression levels of ALS2 and ALS3 mRNA were highest in salivary supernatant without SC + S. sanguis ATCC10556 + oral epithelial cells system,there was significant difference among the three groups(P < 0. 05); In the salivary supernatant containing SC + Susceptible strains + oral epithelial cell system,the expression levels of ALS2 and ALS3 mRNA in salivary supernatant containing SC + S. sanguis ATCC 10556 + oral epithelial cells were the lowest,there was significant difference among the three group(P < 0. 05). Moreover,the expression levels of ALS2 and ALS3 mRNA of three Susceptible strains in containing SC system were significantly lower than those in without SC system(all P < 0. 05).Conclusion the adhesive ability of oral cavity epithelial cells of different susceptible strains is different. The adhesive ability of strain S. sanguis ATCC 10556 was stronger than that of S. mutans UA159 and S. sobrinus OMZ-176. Salivary SC can inhibit the adhesion of susceptible strains to oral epithelial cells,and the strain S. sanguis ATCC 10556 has stronger inhibition ability,it may be related to down regulation of ALS2 and ALS3 gene expression.
引文
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[11] Lecatsas G,Houff S,Macher A,et al. Retrovirus-like particles in salivary glands,prostate and testes of AIDS patients[J]. Proc Soc Exp Biol Med,1985,178(4):653-655
[12]何淼,宋光泰,边专.白色念珠菌ALS基因家族在生物膜形成过程中的差异表达[J].口腔医学研究,2015,31(10):957-960
[2] Han Z,Smith HL. Bacteriophage-resistant and bacteriophage-sensitive bacteria in a chemostat[J]. Math Biosci Eng,2012,9(4):737-765
[3] Warinner C,Rodrigues JFM,Vyas R,et al. Pathogens and host immunity in the ancient human oral cavity[J]. Nature Genetics,2014,46(4):336-344
[4] Hindocha N,Wilson MH,Pring M,et al. Mammary analogue secretory carcinoma of the salivary glands:a diagnostic dilemma[J].Br J Oral Maxillofac Surg,2017,55(3):290-292
[5]张辉,叶美花,俞诚波,等.不同生物状态白色念珠菌对口腔上皮细胞的黏附能力及ALS mRNA表达[J].中国医药导报,2015,12(26):16-19
[6] Murthy AK,Dubose CN,Banas JA,et al. Contribution of polymeric immunoglobulin receptor to regulation of intestinal inflammation in dextran sulfate sodium-induced colitis[J]. J Gastroenterol Hepatol,2006,21(9):1372-1380
[7] Norderhaug IN,Johansen FE,Schjerven H. Regulation of the formation and external transport of secretory immunoglobulins[J].Crit Rev Immunol 1999,19(5/6):481-508
[8] Neutra MR,Kozlowski PA. Mucosal vaccines:the promise and the challenge[J]. Nat Rev Immunol,2006,6(1):148-158
[9] Perrier C,Sprenger N,Corthésy B. Glycans on secretory component participate in innate protection against mucosal pathogens[J]. J Biol Chem,2006,281(20):14280-14287
[10]Phalipon A,Cardona A,Kraehenbuhl JP,et al. Secretory component:a new role in secretory Ig A-mediated immune exclusion in vivo[J]. Immunity,2002,17(1):107-115
[11] Lecatsas G,Houff S,Macher A,et al. Retrovirus-like particles in salivary glands,prostate and testes of AIDS patients[J]. Proc Soc Exp Biol Med,1985,178(4):653-655
[12]何淼,宋光泰,边专.白色念珠菌ALS基因家族在生物膜形成过程中的差异表达[J].口腔医学研究,2015,31(10):957-960