慢传输便秘大鼠肠道菌群的ERIC-PCR指纹图谱分析

目的 应用细菌基因共有重复序列指纹图谱技术,探讨慢传输便秘肠道菌群的变化.方法 选取Wistar大鼠30只,随机分为对照组15只、造模组15只,造模组给予易蒙停灌胃.收集每24 h排出粪便,计数排便粒数,称量粪便质量,及24 h进食量及进水量,通过碳末灌胃测定肠道传输时间.留取造模组与对照组不同时段粪便,提取肠道细菌DNA,进行ERIC-PCR指纹图谱分析.结果 造模组大鼠24 h内排便粒数、排便质量、进食量较对照组明显减少,进水量没有明显差异.造模组大鼠肠道传输时间明显高于对照组.造模组大鼠肠道菌群ERIC-PCR结果优势条带依然存在,但是较对照组及给药前条带增多且亮度增高.结论 易蒙停给药致慢传输便秘大鼠造模成功,造模组大鼠肠道菌群较对照组ERIC-PCR结果可能存在细菌数量及种类增多.

DNA Fingerprinting of Intestinal Flora in Rats with Slow Transit Constipation

Abstract:Objective To analyze the difference of intestinal flora between experimental rats with slow transit constipation （STC） and rats in control group by enter bacterial repetitive intergenic consensus (ERIC)-PCR.Methods Of 30 Wistar rats,15 were randomized to the control group and the other 15 were randomized to model group.We induced STC rats with loperamide by gavage.Fecal pellet numbers,fecal weights were determined.Record the weighing of food and water every 24h.To evaluate gastrointestinal (GI) transit time,two groups of 30 rats were catheters passed into their stomach to allow for administration of a charcoal meal solution via gavage.DNA fingerprinting was set up by using ERIC-PCR technology,and the total difference of their intestinal flora was analyzed.Results Following a 24-day experiment with lopramide,the total fecal mass produced by model group was less than that produced by control rats (4.80±1.48g vs 9.20±.30g,P0.05).After loperamide administration,we observed an increase in the mean GI transit time as compared to the control period (691.60±220.03min vs 437.20±43.89min,P<0.05).The ERIC-PCR electrophoresis showed that there were individual variations of the gut microbiota in rats,while there were common strap in two groups.The diversity of intestinal bacterial composition remarkably increased at the 24th day after giving loperamide in model group.Conclusion Rats model with slow transit constipation were established successfully by feeding loperamide.The diversity of intestinal bacterial composition remarkably increased at the 24th day after giving loperamide.