脱氢表雄酮诱导的多囊卵巢综合征模型大鼠的肠道菌群研究

Gut microbiota of polycystic ovary syndrome model rats induced with dehydroepiandrosterone

目的·分析多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠模型的肠道菌群变化,探讨肠道菌群在其发病过程中可能的作用机制。方法·将20只6周龄清洁级Sprague Dawley雌性大鼠随机分为对照组与实验组,每组10只。实验组大鼠每日皮下注射脱氢表雄酮(dehydroepiandrosterone,DHEA)溶液建立PCOS大鼠模型,对照组皮下注射等量磷酸缓冲盐溶液。4周后检测大鼠卵巢质量、动情周期、卵巢形态学改变;采用Illumina高通量测序技术检测2组大鼠肠道菌群的变化。结果·实验组大鼠的卵巢质量较对照组明显降低(P=0.010)。大鼠动情周期紊乱,同时伴有卵泡异常扩张、卵巢多囊样改变等卵巢形态学改变,提示DHEA成功诱导大鼠出现类PCOS样表现。实验组大鼠肠道菌群在属水平的相对丰度明显改变,其中拟普雷沃菌属(Alloprevotella)及副萨特菌属(Parasutterella)的相对丰度较对照组升高(P=0.040,P=0.009)。LEfSe分析结果提示在2组大鼠之间多种肠道菌存在明显差异,实验组大鼠肠道中α/β/δ-变形杆菌纲(Alphaproteobacteria/Betaproteobacteria/Deltaproteobacteria)、伯克氏菌目(Burkholderiales)、迷踪菌门/纲/目/科(Elusimicrobia/Elusimicrobia/Elusimicrobiales/Elusimicrobiaceae)、Elusimicrobium菌属、副萨特菌属(Parasutterella)及Allobaculum菌属菌群明显富集,而嗜冷杆菌属(Psychrobacter)、Odoribacter菌属及莫拉氏菌科(Moraxellaceae)菌群的丰度显著降低(LDA≥2.0)。结论·PCOS大鼠模型的肠道菌群发生明显改变,多种肠道菌群的丰度发生显著变化,提示PCOS的发生发展过程与肠道菌群的变化有关。

Objective · To explore the changes of gut microbiota in polycystic ovary syndrome (PCOS) model rats and to study the possible role of gut microbiota in the pathological progress of PCOS. Methods · Six-week-old female Sprague Dawley rats were randomly divided into control group and experimental group (n=10 per group). Subcutaneous injection with dehydroepiandrosterone (DHEA) in 0.2 mL phosphate buffer saline (PBS) was adopted to establish PCOS model rats, while the control group rats were subcutaneously injected with the same amount of PBS. After treatment for 4 weeks, the estrous cycle, ovarian weight and morphology were detected. The change of relative abundance of gut microbiota was detected with highthroughput Illumina sequencing technique. Results · Ovarian weight in experimental group was lower than that in control group (P=0.010). The estrous cycle was disrupted and ovarian morphology was greatly changed with enlarged follicles and polycystic ovaries, indicating successful PCOS rat model induced by DHEA. Relative abundance of gut microbiota was significantly altered in genus level, with enrichment of genus Alloprevotella (P=0.040) and Parasutterella (P=0.009) in experimental group. Several kinds of microbial taxa, such as Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Burkholderiale, Elusimicrobia, Elusimicrobiales, Elusimicrobiaceae, and microbial genera Elusimicrobium, Parasutterella and Allobaculum, were remarkably enriched in experimental group, while the abundance of Psychrobacter, Odoribacter and Moraxellaceae were reduced compared with control group revealed by LEfSe analysis (LDA ≥ 2.0). Conclusion · The gut microbiota in PCOS model rats is greatly changed compared with that of control group. Many kinds of microbial taxa varies significantly in abundance, suggesting there might be close association between gut microbiota and occurrence and development of PCOS.