FQ-PCR法分析葛根芩连汤对抗生素相关性腹泻模型肠道肠球菌数量的改变

Analysis of the effect of Gegenqinlian decoction on the changes in the number of intestinal Enterococci in an antibiotic-associated diarrhea model using fluorescent quantitative PCR assay

为了检测葛根芩连汤在治疗抗生素相关性腹泻(AAD)时对模型肠道肠球菌的影响,试验采用氨苄西林灌胃巴马小型猪复制AAD模型,通过制备标准品DNA、构建标准曲线以建立检测肠球菌的FQ-PCR法,利用建立的FQ-PCR法检测对照组(3头)、模型组(3头)、葛根芩连汤7 d治疗组(3头)和葛根芩连汤14 d治疗组(3头)的十二指肠、空肠、回肠、盲肠和结肠内容物中肠球菌的数量。结果表明:通过标准曲线的构建得到肠球菌的标准曲线方程;与对照组相比,模型组肠道内肠球菌含量显著降低(P<0.05);与模型组相比,葛根芩连汤治疗组肠球菌在各个肠段中数量明显上升(P<0.05)。说明FQ-PCR法可以用于AAD模型治疗前后肠道肠球菌的数量检测。

To detect the effect of Gegenqinlian decoction on the intestinal Enterococci in an antibiotic - associated diarrhea （ADD） model treated with Gegenqinlian decoction. Bama miniature pigs were intragastrically administrated with ampicillin to copy ADD model. A fluorescent quanti- tative PCR （FQ -PCR） assay was established by the preparation of standard DNA and the construction of standard curve, and then the num- bers of Enterococci in the contents of duodenum, jejunum, ileum, cecum and colon from the control group （containing 3 pigs）, model group （ containing 3 pigs）, treatment group with Gegenqinlian decoction for 7d （ containing 3 pigs） and treatment group with C-egenqinlian decoction for 14d （containing 3 pigs） were detected using FQ - PCR assay. The results showed that the standard curve equation was constructed through construction of standard curve. The number of intestinal Enterococci in the model group was significantly decreased （ P 〈 0.05 ） compared with a normal control group. The numbers of intestinal Enterococci in each intestinal segment in the treatment groups with Gegenqinlian decoction were obviously increased （ P 〈 0.05 ） compared with the model group. The results indicate that FQ - PCR assay can be used for detecting the number of intestinal Enterococci in the ADD model before and after the treatment.