银翘散治疗儿童风热伤络型过敏性紫癜

Yinqiao Powder in Treatment of Allergic Purpura with Wind-Heat Injured Collateral Syndrome in Children

目的:采用16S rRNA和宏基因组测序方法,比较过敏性紫癜(henoch-sch9nlein purpura,HSP)患儿与健康儿童以及HSP患儿经银翘散治疗后口腔微生物群的组成差异,探究银翘散治疗风热伤络型HSP的可能机制。方法:共纳入初治HSP患儿20例,采集早晨700—800唾液标本为治疗前组,银翘散治疗2周后,再次采集同时段唾液为治疗后组,同时征集20例健康儿童作为健康对照组。采用Illumina NovaSeq测序平台对各组唾液微生物进行16S rRNA高通量测序,通过α多样性和β多样性分析,比较各组菌群物种组成和群落结构的差异,筛选出差异性菌群;采用宏基因组学比较HSP患儿与健康儿童功能基因差异。结果:与健康对照组相比,HSP治疗前组唾液菌群丰富度和多样性下降,但无显著性差异(P>0.05);与治疗前组比较,经银翘散治疗后组唾液菌群丰富度和多样性显著升高(P<0.05)。与健康对照组相比,治疗前组口腔菌群结构无明显差异(P>0.05);与治疗前组比较,治疗后组口腔菌群结构有显著差异(P<0.01)。使用线性判别分析(LDA)效应大小(LEfSe)和MetaStat方法分析,结果显示在门水平上,与健康对照组比较,治疗前组拟杆菌门(Bacteroidota)显著升高;与治疗前组比较,治疗后组Bacteroidota降低。与健康对照组比较,治疗前组放线菌门(Actinobacteriota)降低,与治疗前组比较,治疗后组Actinobacteriota明显升高。在属水平上,与健康对照组比较,治疗前组卟啉单胞菌属(Porphyromonas)、二氧化碳嗜纤维菌属(Capnocytophaga)和颗粒链球菌属(Granulicatella)升高,治疗后组显著低于治疗前组;与健康对照组比较,治疗前组韦荣氏球菌属(Veilonela)降低,治疗后组明显高于治疗前组。HSP患儿与健康儿童存在明显功能基因差异,HSP组患儿口腔细菌群落功能主要集中在信号转导、外源性生物降解和代谢、三大营养物质代谢、抗生素耐药等,而健康儿童口腔菌群与宿主维持生态平衡,主要功能体现在环境适应性免疫和自身免疫的建立。结论:风热伤络型HSP患儿口腔微生态发生了改变,唾液菌群丰富度和多样性下降,经银翘散治疗后恢复了口腔菌群丰富度和多样性,改善了HSP患儿与健康儿童的口腔菌群差异。HSP患儿与健康儿童存在明显功能基因差异。银翘散可能通过改变Porphyromonas、Veilonela、Capnocytophaga、Granulicatella丰富度及相关代谢通路改善口腔微生态,进而达到治疗效果。

Objective:To use 16SrRNA and metagenomic sequencing methods to compare the composition of oral microbiota between children with allergic purpura(henoch-sch nlein purpura,HSP)and healthy children,as well as HSP children treated with Yinqiao Powder,and to explore the possible mechanism of Yinqiao Powder in treatment of of Wind-Heat injured collateral HSP.Methods:A total of 20 children with newly diagnosed HSP were included.The saliva samples were collected from 7 GA6FA 00 to 8 GA6FA 00 in the morning as the pre-treatment group.After Yinqiao Powder treatment for 2 weeks,the saliva was collected again at the same time as the post-treatment group.20 cases were recruited at the same time.Healthy children served as a healthy control group.The Illumina NovaSeq sequencing platform was used to perform high-throughput sequencing of 16SrRNA in each group of saliva microorganisms.Throughα-diversity andβ-diversity analysis,the differences in the species composition and community structure of each group of flora were compared,and the differential flora was screened out;metagenomics was used for comparison Functional gene differences between children with HSP and healthy children.Results:Compared with the healthy control group,the richness and diversity of the salivary flora in the HSP treatment group decreased,but there was no significant difference(P>0.05);compared with the pre-treatment group,the salivary flora in the Yinqiao Powder treatment group Richness and diversity increased significantly(P<0.05).Compared with the healthy control group,there was no significant difference in the oral flora structure of the pre-treatment group(P>0.05);compared with the pre-treatment group,the oral flora structure of the post-treatment group was significantly different(P<0.01).Using the linear discriminant analysis(LDA)effect size(LEfSe)and the MetaStat method,it was shown that at the phylum level,compared with the healthy control group,the Bacteroidota in the pre-treatment group was significantly increased;compared with the pre-treatment group,the post-treatment group Bacteroidota lowered.Compared with the healthy control group,the Actinobacteriota in the pre-treatment group decreased,and compared with the pre-treatment group,the Actinobacteriota in the post-treatment group increased significantly.At the genus level,compared with the healthy control group,the genera of Porphyromonas,Capnocytophaga and Granulicatella in the group before treatment were significantly higher than those in the group after treatment.Before treatment group;compared with the healthy control group,the Veillonella genus(Veilonela)decreased in the pre-treatment group,and was significantly higher in the post-treatment group than in the pre-treatment group.There are obvious functional gene differences between children with HSP and healthy children.The functions of the oral bacterial community in children with HSP are mainly concentrated in:signal transduction,exogenous biodegradation and metabolism,metabolism of three major nutrients,antibiotic resistance,etc.,while healthy children The oral flora maintains the ecological balance with the host,and its main function is reflected in the establishment of environmental adaptive immunity and autoimmunity.Conclusion:The oral microecology of children with Wind-Heat injured type HSP has changed,and the richness and diversity of saliva flora decreased.There are obvious functional gene differences between children with HSP and healthy children.Yinqiao Powder may improve the oral microecology by changing the abundance of Porphyromonas,Veilonela,Capnocytophaga,Granulicatella and related metabolic pathways,and then achieve the therapeutic effect.