辨证治疗小儿轮状病毒肠炎的肠道微生态学研究

Study on intestinal Microecology of Syndrome differentiation in treating Pediatric Rotavirus Enteritis

目的:研究中医辨证治疗小儿轮状病毒肠炎的肠道微生态学变化。方法:选择2015年10月至2017年3月广州中医药大学第一附属医院符合入选标准的6~36个月轮状病毒肠炎患儿共18例,中医辨证分型为湿热证10例,伤食证8例。采用16S rDNA Amplicom测序法检测,分析治疗前后大便菌群的多样性及结构变化。结果:通过香农指数与辛普森指数估算,轮状病毒肠炎在治疗前后,肠道菌群多样性差异无统计学意义(P>0.05),操作分类单元(OTU)水平的差异分析,入选患儿治疗前,沃氏嗜胆菌属、柔嫩梭菌属明显增多,乳酸菌属减少,治疗后,乳酸菌属增多,沃氏嗜胆菌属及柔嫩梭菌属减少,治疗前后比较,差异具有统计学意义(P<0.05);在所有水平上,治疗后的芽孢杆菌纲(bacilli)、乳杆菌科(lactobacillaceae)、乳杆菌属(lactobacillus)、乳杆菌目(lactobacillales)比治疗前明显增加,而变形菌纲(deltaproteobacteria)、弧菌科(desulfovibrionaceae)、瘤胃菌科(ruminococcaceae)、脱硫弧菌目(desulfovibrionales)明显下降,治疗前后比较,差异具有统计学意义(P<0.05);湿热证组治疗前轮状病毒基因浓度为(72.24±29.82)ng·μL^(-1),治疗后为(72.08±22.90)ng·μL^(-1),伤食证组治疗前病毒基因浓度为(80.78±23.19)ng·μL^(-1),治疗后(78.53±15.48)ng·μL^(-1),两组治疗前后比较,差异无统计学意义(P>0.05)。结论:轮状病毒肠炎患儿存在菌群紊乱,双向性的菌群如乳杆菌、大肠杆菌、链球菌、韦荣球菌明显增多,经过治疗后,条件致病菌如梭菌属、弧菌类菌明显减少,乳酸菌属明显增多。中医辨证治疗轮状病毒肠炎的机制可能是通过调整人体肠道菌群,增加有益菌,减少有害菌,平衡菌群来发挥作用。但调节肠道菌群丰度及种类与轮状病毒肠炎的关系,需做进一步的后续研究。

Objective To study on the changes of intestinal microecology in the treatment of rotavirus enteritis in children with TCM syndrome differentiation. Methods Children with rotavirus enteritis who met the criteria for 6 to 36 months were selected. There are 10 cases of dampness and heat syndrome in TCM syndrome differentiation. 8 cases of food injury syndrome,Detection by 16S rDNA Amplicom sequencing. The diversity and structure of fecal flora before and after treatment were analyzed. Results According to Shannon index and Simpson index, there was no significant difference in intestinal bacterial diversity before and after treatment with rotavirus enteritis （P 〉 0.05）. The difference analysis of OTU level showed that before the treatment of all the selected cases, the number of g-Biloplila anf g-Faecalibacterium increased, the number of g-lactohacilus decreased, and after treatment, the number of g-lactobacilus increased, g-Biloplila anf g-Faecalibacterium reduce, and there was significant difference before and after treatment （P 〈 0.05）. At all levels, After treatment, Bacilli, Lactobacillus, Lactobacteriaceae, Lactobacillales higher than before treatment, and Deltaproteobacteria, Desulfovibrionaceae, Ruminococcaceae, Bilophila, Faecalibacterium, Desulfovibrionalesdramatic decline. There was significant difference before and after treatment （P 〈 0.05）. The gene concentration of the former rotavirus in the damp- heat syndrome group was （72.24 ± 29.82） ng/p.L, after treatment was （72.08 ±22.90） ng/μL, The concentration of previrus gene in the group of food injury syndrome was （80.78 ± 23.19） ng/μL, After treatment was （78.53 ± 15.48） ng/μL, There was no significant difference between the two groups before and after treatment （P 〉 0.05）. Conclusion Children with rotavirus enteritis have a disorder of bacterial flora. Bidirectional bacteria such as Lactobacillus, Escherichia, Streptococcus, Veronella, after treatment, conditional pathogenic bacteria as Faecalibacterium, Desulfovibrionale decreased sharply. Lactobacillus increased obviously. The mechanism of TCM syndrome differentiation in treating rotavirus enteritis may be to increase beneficial bacteria and reduce harmful bacteria by adjusting intestinal flora of human body.But the regulation of intestinal flora abundance and the relationship between the species and rotavirus enteritis needs to be further studied.