中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2011年
4期
210-213
,共4页
于丹妮%任志明%韩育植%张文娟
于丹妮%任誌明%韓育植%張文娟
우단니%임지명%한육식%장문연
链球菌,变异%生物膜%显微镜检查,共焦%硫代谢
鏈毬菌,變異%生物膜%顯微鏡檢查,共焦%硫代謝
련구균,변이%생물막%현미경검사,공초%류대사
Streptococcus mutans%Biofilms%Microscopy,confocal%Sulfurmetabolism
目的 采用前期实验已构建成功的luxS基因缺失的变形链球菌突变株,分析luxS基因在口腔变形链球菌的密度感应及硫代谢中的双重作用.方法 在不同硫限定条件培养基(半胱氨酸、蛋氨酸、标准株无菌上清液、S-腺苷高半胱氨酸、S-腺苷甲硫氨酸,空白对照为脑心浸液培养基)中分别对不同培养时间(24、48 h)的变形链球菌标准株和突变株的生长进行吸光度值(A值)的测定与分析,通过激光扫描共聚焦电子显微镜观察测定并比较不同培养条件下两菌株生物膜厚度的差异.结果 在半胱氨酸培养基中培养48 h,突变株生长A值及生物膜厚度均有所增长,分别为(1.301±0.009)和(45.009±0.429)μm,但未及标准株水平(P＜0.05);在蛋氨酸及S-腺苷高半胱氨酸培养基中培养24 h,突变株的生长A值及生物膜厚度与空白对照相比均有所补充,在蛋氨酸培养基中突变株的生长A值及生物膜厚度分别为(0.448±0.028)和(37.068±2.392)μm,在S-腺苷高半胱氨酸培养基中,突变株的生长A值及生物膜厚度分别为(0.460±0.005)和(27.343±1.107)μm,但均未及标准株水平(P＜0.05),48 h时达到标准株水平;在标准株上清液培养基中突变株的生长A值及生物膜厚度均明显增高且超过标准株水平;在S-腺苷甲硫氨酸培养基中两菌株的生长A值及生物膜厚度与空白对照相比均有不同程度的降低.结论 luxS基因在变形链球菌中不但具有密度感应功能,在硫代谢方面也发挥着一定作用.
目的 採用前期實驗已構建成功的luxS基因缺失的變形鏈毬菌突變株,分析luxS基因在口腔變形鏈毬菌的密度感應及硫代謝中的雙重作用.方法 在不同硫限定條件培養基(半胱氨痠、蛋氨痠、標準株無菌上清液、S-腺苷高半胱氨痠、S-腺苷甲硫氨痠,空白對照為腦心浸液培養基)中分彆對不同培養時間(24、48 h)的變形鏈毬菌標準株和突變株的生長進行吸光度值(A值)的測定與分析,通過激光掃描共聚焦電子顯微鏡觀察測定併比較不同培養條件下兩菌株生物膜厚度的差異.結果 在半胱氨痠培養基中培養48 h,突變株生長A值及生物膜厚度均有所增長,分彆為(1.301±0.009)和(45.009±0.429)μm,但未及標準株水平(P＜0.05);在蛋氨痠及S-腺苷高半胱氨痠培養基中培養24 h,突變株的生長A值及生物膜厚度與空白對照相比均有所補充,在蛋氨痠培養基中突變株的生長A值及生物膜厚度分彆為(0.448±0.028)和(37.068±2.392)μm,在S-腺苷高半胱氨痠培養基中,突變株的生長A值及生物膜厚度分彆為(0.460±0.005)和(27.343±1.107)μm,但均未及標準株水平(P＜0.05),48 h時達到標準株水平;在標準株上清液培養基中突變株的生長A值及生物膜厚度均明顯增高且超過標準株水平;在S-腺苷甲硫氨痠培養基中兩菌株的生長A值及生物膜厚度與空白對照相比均有不同程度的降低.結論 luxS基因在變形鏈毬菌中不但具有密度感應功能,在硫代謝方麵也髮揮著一定作用.
목적 채용전기실험이구건성공적luxS기인결실적변형련구균돌변주,분석luxS기인재구강변형련구균적밀도감응급류대사중적쌍중작용.방법 재불동류한정조건배양기(반광안산、단안산、표준주무균상청액、S-선감고반광안산、S-선감갑류안산,공백대조위뇌심침액배양기)중분별대불동배양시간(24、48 h)적변형련구균표준주화돌변주적생장진행흡광도치(A치)적측정여분석,통과격광소묘공취초전자현미경관찰측정병비교불동배양조건하량균주생물막후도적차이.결과 재반광안산배양기중배양48 h,돌변주생장A치급생물막후도균유소증장,분별위(1.301±0.009)화(45.009±0.429)μm,단미급표준주수평(P＜0.05);재단안산급S-선감고반광안산배양기중배양24 h,돌변주적생장A치급생물막후도여공백대조상비균유소보충,재단안산배양기중돌변주적생장A치급생물막후도분별위(0.448±0.028)화(37.068±2.392)μm,재S-선감고반광안산배양기중,돌변주적생장A치급생물막후도분별위(0.460±0.005)화(27.343±1.107)μm,단균미급표준주수평(P＜0.05),48 h시체도표준주수평;재표준주상청액배양기중돌변주적생장A치급생물막후도균명현증고차초과표준주수평;재S-선감갑류안산배양기중량균주적생장A치급생물막후도여공백대조상비균유불동정도적강저.결론 luxS기인재변형련구균중불단구유밀도감응공능,재류대사방면야발휘착일정작용.
Objective To investigate the function of luxS in sulfurmetabolism of Streptococcus mutans (Sm). Methods The growth with absorbency(A) of the standards and mutant strains was measured and analyzed in the sulfur-limited defined medium at different periods. The laser scanning confocal microscopy( LSCM ) was used to observe and compare the biofilm thickness of the two kinds of strains at different culture conditions. Results The significant increases in the thickness of mutant strain biofilm and its growth were observed after the addition of cysteine, but did not reach the standards strain levels ( P ＜0. 05). The growth and the biofilm thickness of the mutant strains were ( 1. 301 ±0. 009) and (45.009 ±0. 429) μm. When methionine and S-adenosylhomocysteine of certain concentrations were respectively added, the biofilm thickness and the growth of mutant strain were raised but did not reach the level of the standards strain at 24 h( P ＜0. 05 ), but at 48 h they did. When the methionine was added in the mutant strains for 24 h, the biofilm thickness and the growth of mutant strain were (0. 448 ± 0. 028 ) and ( 37. 068 ± 2. 392 ) μm, as for the adding of S-adenosylhomocysteine were (0. 460 ± 0. 005 ) and ( 27. 343 ±1. 107 ) μm. When adding the supernatant fluid of standard strains, the biofilm thickness and the growth levels of mutant strain were much higher than those of the standards strain. The biofilm thickness and growth of both kinds of strains decreased after the addition of S-adenosylmethionine. Conclusions luxS gene plays not only a role in quorum sensing but also a role in sulfurmetabolism.
