关键词: 肠道损伤, 红花清肝13味丸, 转录组测序, 信号通路, 差异基因

Abstract: [Objective] The aim of the present study was to screen the important signaling pathways and genes involved in carbon tetrachloride （CCl₄） induced intestinal injury in mice and in the treatment process with a traditional Mongolian medicine Honghua Qinggan 13 Flavor Pills. [Method] A total of 18 male C57BL/6 mice aged 6-8 weeks and weighing 20-22 g were randomly divided into three groups, a blank control group （n=6）, a model group （n=6）, and a treatment group （n=6）. The blank control group and model group were gavaged with physiological saline once a day at a dose of 10 mL/（kg·BW） for 7 consecutive days. The treatment group was gavaged with Honghua Qinggan 13 Flavor Pills once a day at a dose of 616.5 mg/（kg·BW） for 14 consecutive days. Four hours after the last gavage, the blank control group was intraperitoneally injected with olive oil, while both the model group and treatment group were intraperitoneally injected with CCl₄ and olive oil mixture （CCl₄∶olive oil=1∶4, V/V） at a dose of 10 mL/（kg·BW）. On the second day following the intestinal injury treatment, the duodenal tissues of the mice in the three groups were collected for high-throughput transcriptome sequencing. Using the obtained sequencing data, the differentially expressed genes were predicted, functionally analyzed, as well as KEGG signaling pathway enriched. The gene groups enriched in the significantly down regulated signaling pathway after the treatment with Honghua Qinggan 13 Flavor Pills were screened, and interaction analysis was performed with the target genes of Honghua Qinggan 13 Flavor Pills. [Result] ① Transcriptome sequencing analysis showed that compared with the blank control group, the model group and the treatment group obtained 1 419 and 2 875 differentially expressed genes, respectively. Compared with the model group, the treatment group obtained 2 288 differentially expressed genes, of which 1 139 were up regulated and 1 149 were down regulated. KEGG signaling pathways enrichment on the differentially expressed genes in the model group found that the activated signaling pathways included osteoclast differentiation, B cells, chemokines, inflammatory bowel disease, JAK-STAT, etc. KEGG signaling pathways enrichment on the differentially expressed genes in the treatment group found that the activated signaling pathways included FOXO, mTOR, longevity regulation, etc, while the inhibited signaling pathways included osteoclast differentiation, cell apoptosis, tumor necrosis factor, B cells, rectal and colorectal carcinoma, etc. ②Four genes with significantly different expression levels, JUN, STAT1, NFKBIA and FOS, were found in the transcriptome data via the gene groups enrichment in the significantly inhibited signaling pathway in the treatment group and interaction analysis between the target genes of Honghua Qinggan 13 Flavor Pills. [Conclusion] The main signaling pathways involved in CCl₄ induced intestinal injury in mice and the treatment process of Honghua Qinggan 13 Flavor Pills included chemokines, inflammatory bowel disease, JAK-STAT and longevity regulation, and the main genes included JUN, STAT1, NFKBIA and FOS. Our results provided references for clarifying the molecular mechanisms of prevention and treatment of intestinal injury with Honghua Qinggan 13 Flavor Pills.

Key words: intestinal injury, Honghua Qinggan 13 Flavor Pills, transcriptome sequencing, signaling pathway, differential gene