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目的采用多聚酶链反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradientgel electrophoresis,PCR-DGGE)技术研究中药复方人参乌梅汤加味对腹泻大鼠肠道菌群构成特征的影响。方法 48只SD大鼠随机抽取12只为空白组,其余36只参照文献方法以复合因素制作腹泻模型。造模成功后随机分为模型组、西药组、中药组,每组12只。造模21d开始给药干预,空白组和模型组予生理盐水,其余各组分别给予妈咪爱、人参乌梅汤加味,均1次/d,连续给药7d。收集各组大鼠粪便排泄物,提取粪便细菌总基因组DNA,PCR扩增细菌16Sr DNA基因V3可变区,DGGE方法检测PCR产物。对优势条带进行回收测序,鉴定菌种。结果药物处理后腹泻模型大鼠肠道微生物相对丰度最高的为厚壁菌门类细菌,其次为拟杆菌门和变形菌门类细菌。人参乌梅汤加味治疗后大鼠肠道厚壁菌门类细菌含量显著增加(P<0.01),而肠道拟杆菌门类细菌含量显著降低(P<0.01),肠道变形菌门类细菌含量无显著变化(P>0.05)。与空白组相比,经人参乌梅汤加味治疗后大鼠菌群较之西药组更接近空白组。结论人参乌梅汤加味较之西药对肠道菌群平衡更具有良性的调控作用,可能通过影响肠道菌群结构,纠正菌群紊乱,进而发挥生津止泻效应。
Objective To investigate the effect of traditional Chinese medicine compound gmei wu tumei on the composition of intestinal microflora in diarrhea rats by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Methods 48 Sprague - Dawley rats were randomly divided into two groups, of which 12 were randomly selected. After the success of modeling were randomly divided into model group, western medicine group, traditional Chinese medicine group, each group of 12. At the beginning of model 21d, drug administration was given, saline and saline were given to the blank group and the model group, respectively. The other groups were given mommy love and ginseng wu mei tong respectively, once a day for 7 days. The faecal excrement of rats in each group was collected, total genomic DNA of fecal bacteria was extracted, the V3 variable region of bacterial 16Sr DNA gene was amplified by PCR, and PCR products were detected by DGGE. The dominant bands were recovered and sequenced to identify strains. Results The highest relative intestinal microbiota of drug-treated diarrhea model rats was F-type bacteria, followed by Bacteroidetes and Proteobacteria. The contents of bacteria in the intestinal bacteria were significantly increased (P <0.01), while the contents of bacteria in the intestinal bacteria were significantly decreased (P <0.01), while the contents of bacteria in the intestinal bacteria were not significantly changed (P> 0.05). Compared with the blank group, the ginseng Wumei Decoction treatment rat bacterial flora than the Western medicine group closer to the blank group. Conclusion Ginseng Wu Mei Decoction has a more benign regulation of gut flora balance than western medicine, which may correct the flora disruption by influencing gut flora structure, and then exert the anti-diarrhea effect of Sheng Jin.