双歧杆菌小鼠肠道定植的优化

Optimize on colonization of Bifidobacterium

目的评价自制的小鼠灌肠器灌肠效果,并探讨双歧杆菌定植的优化方式,更好地建立双歧杆菌在结肠的定植模型。方法选用SPF级C57BL小鼠40只,每组10只,随机分成4组:正常对照组,常规灌胃组,市售灌肠组,自制灌肠组。从市面不同的材料中筛选小鼠灌肠管,并设计小鼠专用灌肠器。记录不同灌注方式时隐血试验等一般指标检测。RT-PCR检测4组脾脏TLR2 mRNA表达水平。免疫组化观察颈动脉血管HGF蛋白水平。体外模拟不同pH值检测双歧杆菌定植HT-29细胞的粘附水平。结果自制的灌肠器在小鼠灌肠操作过程中明显优于市售灌肠器;自制灌肠组隐血试验明显低于市售灌肠组(P<0.01);RT-PCR结果显示,自制灌肠组效果最强,市售灌肠组次之,常规灌胃组一般,但均高于正常对照组;HGF的免疫组化显示,自制灌肠组表达最强;粘附试验显示双歧杆菌在pH 7.0环境中粘附能力优于pH 4.5。结论双歧杆菌在结肠定植采用灌肠方式优于灌胃方式;自制灌肠器优于市售灌肠器;优化灌肠液,采用自制的灌肠器灌肠同时优化灌肠操作,能够在小鼠结肠建立较好的双歧杆菌定植模型。

Objective To evaluate the colonization effect of Bifidobacterium by home-made enteroclysm on C57BL mice, and study how to optimize on colonization effect of Bifibacterium to the intestinal mucosa. Methods C57BL mice in SPF grade were divided into four groups from Chongqing University of Medical Sciences. Mice in control group（ n = 10） , mice in convention intragastric administration group（ n = 10） and in market enteroclysm and home-made enteroclysm group （ n = 10） were received bifidobacteria 1×10^9 CFU/day for 30 days and collected information. Mice were sacrificed and collected specimen to study TLR2 mRNA by RT-PCR and HGF by immunohistochemistry. Moreover, the adhesive effect of Bifidobacterium were measured in pH 7.0 and 4.5. Results The levels of TLR2 mRNA and HGF significantly increased in enteroclysm compared to intragastric administration. The colonization effect of Bifidobacterium to HT-29 cells showed significantly increased in pH 7.0 compared to pH 4.5. Conclusions Enteroclysm is better compared to intragastric administration and home-made enteroclvsm is better than others in mice to optimize on colonization effect.