探讨不同浓度的清郁和降汤对反流性食管炎大鼠肠道菌群的影响

Different concentrations of Qingyu Hejiang Decoction regulate intestinal flora in rats with reflux esophagitis

摘要:

【目的】观察不同浓度的清郁和降汤对反流性食管炎（reflux esophagitis，RE）的治疗效果，并探讨其对肠道菌群的影响。【方法】将36只健康雄性SD大鼠随机分为6组，其中1组为假手术组，剩余5组大鼠采用“前胃结扎+外置幽门部分结扎术”手术造模方法建立反流性食管炎模型。造模2周后将术后全部存活的30只大鼠随机分成对照组、中药高剂量组（予高剂量清郁和降汤）、中药中剂量组（予中剂量清郁和降汤）、中药低剂量组（予低剂量清郁和降汤）、西药组（予泮托拉唑钠肠溶胶囊+枸橼酸莫沙比利分散片+复方嗜酸乳杆菌片），每组6只。于术后第15天开始灌胃，其中假手术组及对照组予蒸馏水灌胃，其他组分别给予相应的药物灌胃，持续灌胃14 d后将所有大鼠处死后进行取材。以苏木精-伊红（hematoxylin-eosin，HE）染色观察大鼠食管组织的病理学改变；采用16S rRNA基因高通量测序检测其肠道黏膜的菌群构成。【结果】反流性食管炎大鼠存在着较为明显的肠道菌群结构变化及肠道菌群多样性较低的情况，B组（对照组）中厚壁菌门和拟杆菌门占比减少，变形菌门占比增多，且假单胞菌属、青枯菌属等细菌增多。低、中、高3种浓度的清郁和降汤均能够提升RE大鼠的肠道菌群多样性，增加拟杆菌门及厚壁菌门，降低变形菌门的占比，从属水平上看，清郁和降汤能够提升大鼠肠道中拟杆菌属、乳杆菌属、瘤胃球菌属、颤螺旋菌属、双歧杆菌属和狄氏副拟杆菌属等益生菌占比。以D组（中剂量组）对大鼠肠道菌群多样性提升最为明显，其效果最接近假手术组。特征微生物方面，B组以变形菌门为特征性微生物，D、E两组出现了放线菌门及拟杆菌门下属细菌为特征微生物的情况，在门水平与F组相同。【结论】清郁和降汤能够有效地治疗反流性食管炎，其机制可能与改变RE大鼠的肠道菌群结构、减少有害菌、提升益生菌的占比和改善肠道菌群多样性有关。

Abstract:

[Objective] To observe the effects of different concentrations of Qingyu Hejiang Decoction on reflux esophagitis (RE) and intestinal flora.[Methods] Thirty-six healthy male SD rats were randomly assigned into six groups. Except the sham operation group, the remaining five groups received anterior gastric ligation+external partial pyloric ligation for the modeling of RE. Two weeks after modeling, all the 30 surviving rats were randomly assigned into a sham operation group, a control group, a western medicine (pantoprazole sodium enteric-coated capsules+mosapride citrate dispersible tablets+compound lactobacillus acidophilus tablets) group, and high-, medium-, and low-dose Qingyu Hejiang Decoction groups (n=6). The rats in the sham operation group, the control group, and other groups were administrated with distilled water, distilled water, and corresponding drugs, respectively, by gavage. After 14 days of intervention, all the rats were sacrificed for the collection of tissue samples. The pathological changes of the esophageal tissue were observed via hematoxylin-eosin (HE) staining, and 16S rRNA gene high-throughput sequencing was employed to determine the bacterial composition in the intestinal mucosa. [Results] The RE rats showed significant changes and low diversity of intestinal flora. The control group showed decreased relative abundance of Firmicutes and Bacteroidetes, increased relative abundance of Proteobacteria, and increased Pseudomonas and Ralstonia. Low-, medium-, and high-dose Qingyu Hejiang Decoction increased the diversity of intestinal flora in RE rats. Specifically, they increased the relative abundance of Firmicutes and Bacteroidetes and reduced that of Proteobacteria. At the genus level, Qingyu Hejiang Decoction increased the relative abundance of probiotics such as Bacteroides, Lactobacillus, Ruminococcus,Oscillospira, Bifidobacterium, and Parabacteroides in the intestinal tract of RE rats. Moreover, the medium-dose group demonstrated the best performance, with the intestinal microflora diversity close to that of the sham operation group. Proteobacteria was the characteristic microorganisms in the control group, while Actinomycetes and Bacteroidetes were the characteristic microorganisms in the medium- and high-dose Qingyu Hejiang Decoction groups, which were the same as those in the western medicine group. [Conclusion] Qingyu Hejiang Decoction can effectively treat RE by reshaping the intestinal flora structure (reducing harmful bacteria, increasing probiotics, and increasing the diversity of intestinal flora).