Objective:To explore the intestinal flora diversity and endogenous metabolites in fecal of rats with cold coagulation and blood stasis syndrome

and to investigate gut flora

metaboites and metabolic pathways closely related to cold coagulation and blood stasis syndrome. Method:The rats model of cold coagulation and blood stasis syndrome induced by ice-water bath was established

16S rRNA high-throughput gene sequencing coupled with UPLC-TOF-MS based on fecal metabonomics were used to analyze relativity between intestinal flora and endogenous biomarkers. Result:Based on Illumina MiSeq platform

V4 region was selected as the ideal variable regions.Compared with the blank group

it was found that Firmicutes was markedly up-regulated in model group(P<0.05) with Bacteroidetes decreased(P<0.01)

there were 23 genus with significant differences.Based on fecal metabonomics technology

seven biomarkers were identified in model rats

of which 6 metabolites was markedly up-regulated except for docosapentaenoic acid.Three key metabolic pathways including histidine metabolism

biosynthesis of valine

leucine and isoleucine

biosynthesis of pantothenic acid and coenzyme A(CoA) were founded.There was a strong correlation between gut microbiota genera and fecal metabolites. Conclusion:It is suggested that 16S rRNA high-throughput gene sequencing combined with metabolomics can be further applied to assess pathogenesis of cold coagulation and blood stasis syndrome.