CAJ | 학술논문

目的探讨不同龋敏感儿童口腔变异链球菌 HtrA mRNA和蛋白质的表达情况与乳牙龋坏程度和 HtrA的关系。方法以高龋、中龋和无龋儿童口腔中分离得到的变异链球菌为实验菌株，复苏，增菌，分离纯化核酸，采用逆转录 PCR（RT-PCR）法，提取变异链球菌临床分离株总RNA ，凝胶电泳检测RNA完整性，合成cDNA ，PCR扩增，将扩增产物凝胶成像系统下观察记录结果，将目的基因HtrA及内参的电泳图像利用凝胶定量分析软件Gel-Pro analyzer 4．0进行灰度扫描，分析计算基因相对表达值；采用蛋白质印迹（Western blot）法，验证变异链球菌临床分离株总蛋白，结果用Bio-Rad凝胶摄像分析系统扫描入计算机中，凝胶定量分析软件Quantity One 4．4．0分析其灰度值，计算蛋白的相对表达水平。结果高龋、中龋及无龋儿童口腔变异链球菌临床分离株HtrA mRNA的表达和 HtrA蛋白的表达均存在差异，从高到低为高龋组、中龋组、无龋组，差异有统计学意义（P＜0．05）。结论不同龋敏感儿童口腔变异链球菌临床分离菌株HtrA mRNA的表达和HtrA蛋白的表达均存在差异，龋敏感性越高，HtrA mRNA的表达和 HtrA蛋白的表达越高。
목적탐토불동우민감인동구강변이련구균 HtrA mRNA화단백질적표체정황여유아우배정도화 HtrA적관계。방법이고우、중우화무우인동구강중분리득도적변이련구균위실험균주，복소，증균，분리순화핵산，채용역전록 PCR（RT-PCR）법，제취변이련구균림상분리주총RNA ，응효전영검측RNA완정성，합성cDNA ，PCR확증，장확증산물응효성상계통하관찰기록결과，장목적기인HtrA급내삼적전영도상이용응효정량분석연건Gel-Pro analyzer 4．0진행회도소묘，분석계산기인상대표체치；채용단백질인적（Western blot）법，험증변이련구균림상분리주총단백，결과용Bio-Rad응효섭상분석계통소묘입계산궤중，응효정량분석연건Quantity One 4．4．0분석기회도치，계산단백적상대표체수평。결과고우、중우급무우인동구강변이련구균림상분리주HtrA mRNA적표체화 HtrA단백적표체균존재차이，종고도저위고우조、중우조、무우조，차이유통계학의의（P＜0．05）。결론불동우민감인동구강변이련구균림상분리균주HtrA mRNA적표체화HtrA단백적표체균존재차이，우민감성월고，HtrA mRNA적표체화 HtrA단백적표체월고。
Objective To evaluate the relationship between mRNA and protein expression of HtrA and Streptococcus mutans i-solated from the children with different caries experience and to provide the theoretical and experimental basis on prediction of dent-al caries in deciduous teeth .Methods The strains of Streptococcus mutans isolated from children with different carious experiences in the preliminary experiments were divided into three groups :high caries-susceptible group ,middle caries-susceptible group ,caries-free group .All strains were reanimated on the agar plate of MS ,and after smear pure culture examination ,typical bacteria were in-cubated in BHI ,then purified nucleic acid and extracted all the RNA of streptococcus mutans by reverse transcription PCR and de-tected it by agarose gel electrophoresis integrality .Synthetic cDNA and take further PCR amplification with cDNA products .Ob-serve records results by Gel imaging system .HtrA of target gene and electrophoresis image were gray scan by Gel quantitative soft-ware Gel-Pro analyzer 4 .0 was used to analyze relative expression value of gene .After purifying protein ,collected total protein of Streptococcus mutans strains by Western Blot method ,then tested the concentration of total protein sample .The results of Chemilu-minescence imaging were scanned into computer by Bio-Rad analyzing system ,calculated the gray value by software Quantity One 4 .4 .0 which showed the relative expression level of protein .Results There were significant differences in HtrA mRNA and protein expression of different Streptococcus mutans isolated from the children with different caries susceptibility .high caries-susceptible group> middle caries-susceptible group> caries-free group (P< 0 .05) .Conclusion There were significant differences in HtrA mRNA and protein expression of different Streptococcus mutans isolated from the children with different caries susceptibility .The higher caries susceptibility the group was ,the more HtrA mRNA and protein the strain express .