【作者】 杨光；

【导师】 辛毅；

【作者基本信息】 大连医科大学 ， 生物化学与分子生物学， 2014， 硕士

【摘要】 目的：酸浆多糖为中药锦灯笼的主要成分，具有抗肿瘤、抗衰老、提高机体免疫力的功效。本课题采用PCR-DGGE和Real-time方法对不同剂量的酸浆多糖调节抗生素所致肠道菌群的失调进行研究。方法：选用SPF级BALB/c小鼠，随机分成6组，第1组为空白对照，2-6组每天灌服50㎎/㎏盐酸左氧氟沙星7天，于实验第7天收集每只小鼠粪便。实验第8天，2-4组分别灌服不同剂量（10㎎/ml、20㎎/ml、40㎎/ml）的酸浆多糖，第5组为阳性对照（丽珠肠乐），在第14天，收集每只小鼠的粪便，提取粪便中细菌基因组DNA，再应用PCR-DGGE技术获得小鼠肠道菌群分子的指纹图谱，进行相似性、多样性分析，选取优势条带切胶、克隆测序，对序列进行分析。进而应用实时定量PCR技术对优势菌进行定量分析。结果：酸浆多糖组小鼠与空白对照组的小鼠粪便细菌DGGE图谱有明显差异(P <0.05)。Real-time PCR结果显示：与空白对照组相比，酸浆多糖组双歧杆菌的数量均显著增加(P <0.05)。酸浆多糖表现出刺激益生菌的生长，同时抑制体外病原菌的生长的效果。随着体内酸浆多糖浓度的增加，乳杆菌属的物种的数量显著增加。结论：酸浆多糖能够促进肠道益生菌的生长，对肠道菌群失调具有调节作用。更多还原

【Abstract】 Objective: PPSB is the main ingredient of Chinese medicine Calyx seu FructusPhysalis, has effect on anti-tumor, anti-aging, and improving immunity.In this projectwe apply PCR-DGGE and Real-time to explore the effect of PPSB with variousconcentrations on the regulation of Mouse intestinal microflora imbalance.Methods: BALB/c(SPF), randomly divided into6group，Group1as the controlgroup, Group2-6Irrigate50mg/kg of levofloxacin hydrochloride daily for7days,mouse stool was collected at day7. at day8, groups2-4were fed with different doses(10mg/ml,20mg/ml,40mg/ml) of acid slurry polysaccharide, Group5as apositive control (Lizhuchangle), at day14, stool of each mouse was collected, extractgenomic DNA, then gain the molecular fingerprints of the intestinal flora byPCR-DGGE technique, the similarity,sequence diversity and the advantages of thesuperiority strip were analysised. the quantitative of dominant bacterias were furtheranalysised by real-time PCR.Results: DGGE profiles of mice fecal bacterial were significantly different (P<0.05) between PPSB group and control group, Real-time PCR results showedthat,Compared with the control group, number of bifidobacteria increased significantly(P <0.05). PPSB group of Physalis polysaccharides can promote the growth ofprobiotics, while inhibiting the growth of pathogenic bacteria in vitro. With increasingconcentration of polysaccharides in vivo, the number of Lactobacillus speciesincreased significantly. Conclusion: Physalis polysaccharide can promote the growth of probiotics andhas an effect on the regulation of intestinal microflora imbalance.更多还原