痛泻要方对肝郁脾虚型胃溃疡大鼠肠道菌群的影响

Effects of Tongxie Yaofang on intestinal flora of rats with liver depression and spleen deficiency type gastric ulcer

目的探究痛泻要方对肝郁脾虚型胃溃疡大鼠肠道菌群的影响。方法将18只SD大鼠随机分为3组:空白对照组、模型组和痛泻要方组(8.32g/kg)。采用中医病因多因素复合模拟(第1~14天)和乙酸法(第15天)建立肝郁脾虚型胃溃疡大鼠模型。各组大鼠灌服相应溶液(第16~30天)。ELISA法检测各组大鼠的血清胃泌素(GAS)、5-羟色胺(5-HT),HE染色观察胃组织病理改变,采集大鼠粪便,提取肠道菌群DNA,使用NovaSeq 6000测序仪对16S rDNA的V3-V4区测序分析,分析其多样性及结构差异。结果与模型组相比,痛泻要方组血清中GAS和5-HT水平均显著升高(P<0.01);大鼠肠道菌群的α多样性指数中的chao1、observed species、shannon指数均明显升高(P<0.01、P<0.01、P<0.05),其肠道菌群中放线菌门的相对丰度显著降低(P<0.01),考拉杆菌属的相对丰度显著降低(P<0.01);痛泻要方组富集了普雷沃氏菌科Ga6A1组。结论痛泻要方发挥抗肝郁脾虚型胃溃疡的作用与调节肠道菌群多样性与结构密切相关。

Objective To investigate the effect of Tongxie Yaofang on the intestinal flora of rats with liver depression and spleen deficiency type gastric ulcer.Methods Eighteen SD rats were randomly divided into three groups:blank control group,model group and Tongxie Yaofang group(8.32g/kg).The rat model of gastric ulcer of liver-depression and spleen-deficiency type was established by using the multifactorial composite simulation of traditional Chinese medicine etiology(day 1~14)and acetic acid method(day 15).The rats in each group were dosed with the corresponding solutions(days 16~30).Serum gastrin(GAS)and 5-hydroxytryptamine(5-HT)were detected by ELISA in each group,and HE staining was performed to observe the pathological changes in gastric tissues.The feces of rats were collected,and DNA of intestinal flora was extracted,and the V3-V4 region of the 16S rDNA was sequenced using a NovaSeq 6000 sequencer,and analyzed for diversity and structural differences.Results Compared with the model group,the blood serum levels of GAS and 5-HT in the Tongxie Yaofang group were significantly higher(P<0.01);theαdiversity indices of rat intestinal flora of chao1,observed species,and shannon indices were all significantly higher(P<0.01,P<0.01,P<0.05),and the relative abundance of Actinobacteria in its intestinal flora was significantly lower(P<0.01),and the relative abundance of Phascolarctobacterium was significantly lower(P<0.01);the Tongxie Yaofang group was enriched with Prevotellaceae Ga6A1 group.Conclusion The effect of Tongxie Yaofang in counteracting gastric ulcer caused by liver depression and spleen deficiency is closely related to the regulation of the diversity and structure of intestinal flora.