Abstract: OBJECTIVE To analyze the characteristics of intestinal flora in rats with chronic obstructive pulmonary disease (COPD) of lung and spleen deficiency based on 16S rRNA gene sequencing technology. METHODS Totally 30 Wistar rats were randomly divided into the blank control group and the model group, with 15 rats in each group. The rats of the model group were treated with fumigation plus cold senna leaf infusion to establish COPD models of lung and spleen deficiency. The stool samples were collected from the two groups after the modeling. 16S rRNA sequencing was carried out for the intestinal flora, and the changes of structure, diversity and abundance of intestinal flora were observed. RESULTS The result of Wayne diagram based on operational taxonomic unit (OTU) showed the number of OUT of the model group was less than that of the blank control group. The analysis of structure of flora showed that Firmicutes was dominant in the blank control group, and Bacteroides was dominant in the model group. At the genus level, Lactobacillus was dominant among Firmicutes in the blank control group, and Bacteroides was dominant in the model group, and the counts of Lactobacillus of the model group were less than those of the blank control group. Principal coordinate analysis (PCoA) indicated that there was significant difference in the composition of intestinal flora between the two groups; at the phyla level, the Actinomycetes counts of the model group were less than those of the blank control group, while the Proteus and Desulfobacter counts of the model group were more than those of the blank control group. At the genus level, the Bacteroides, Rombucella, Trichospirillum and Parabacteroide counts of the model group were more than those of the blank control group, while the Prevotella, Clostridium, Brucella and Rumen coccus counts of the model group were less than those of the blank control group. Diversity of flora showed that Shannon index and sobs index of the model group were lower than those of the control group. CONCLUSION The structure, relative abundance and diversity of the intestinal flora have changed in the rats with COPD of lung and spleen deficiency.