卵巢过度刺激综合征大鼠肠道菌群高通量测序初步探索

High-throughput sequencing of intestinal flora of rats with ovarian hyperstimulation syndrome

目的分析比较卵巢过度刺激综合征大鼠肠道菌群的结构特征,探讨外源激素的应用对大鼠肠道菌群的影响。方法通过皮下注射不同剂量的激素构建卵巢过度刺激综合征组(超促排剂量)、超促排卵组(正常促排剂量)和生理盐水对照组的大鼠模型,并收集大鼠的肠道内容物,提取参试样本中细菌总DNA,进行16S rDNA基因高通量测序。结果高通量测序结果显示,3组大鼠肠道内容物菌群的测序覆盖指数均达到0.993以上;随着外源激素用量的增加,肠道菌群结构的多样性亦随之提高;3组大鼠菌群结构在门水平上主要由厚壁菌门(Firmicutes)、疣微菌门(Verrucomicrobia)和拟杆菌门(Bacteroidetes)3个门组成;在属水平上主要由乳酸杆菌属(Lactobacillus)、艾克曼菌属(Akkermansia)、肠球菌(Enterococcus)和梭状芽胞杆菌XlVa(Clostridium XlVa)4个属组成,其中艾克曼菌属(Akkermansia)的相对丰度随着外源激素用量增加而下降,相反的肠球菌(Enterococcus)和梭状芽胞杆菌XlVa(Clostridium XlVa)则升高。结论外源激素的使用会改变大鼠肠道菌群的结构平衡状态,菌群的多样性提高;引起部分益生菌相对丰度下降,而致病菌的相对丰度增加,影响机体的过度刺激反应。

Objective To compare the characteristics of intestinal flora among rats with ovarian hyperstimulation syndrome and super stimulate ovulation, and normal rats, thus to investigate the effect of exogenous hormone on intestinal flora. Methods The ovarian hyperstimulation syndrome and the super stimulate ovulation rat models were constructed through subcutaneous injection of different doses of the hormone. The intestinal contents were collected, and bacterial whole DNA were extracted. The regions V3-V4 of 16 s rDNA gene were sequenced by high-throughput sequencing.Results The high-throughput sequencing coverage index of the intestinal contents of the three groups were all greater than 0.993. With the increase of exogenous hormone, the diversity of intestinal microflora was also increased. The predominant 3 phyla in these three groups were Firmicutes, Verrucomicrobia and Bacteroidetes. On the genus level, the predominant 4 genus were Lactobacillus, Akkermansia, Enterococcus and Clostridium XlVa. With the increase of exogenous hormones, the relative abundance of Akkermansia was decreased, while Enterococcus and Clostridium XlVa were increased. Conclusion The exogenous hormones injection alters the homeostasis and increase the diversity of intestinal flora in rats. It reduces the relative abundance of specific probiotics, while increases the relative abundance of pathogenic bacteria, resulting in corresponding overstimulation response.