基于16S rRNA测序与非靶向代谢组学探究光甘草定抗结直肠癌作用OA

Objective By constructing a MC38 tumor-bearing mouse model,combined with 16S rRNA high-throughput sequencing and non-targeted metabolomics techniques,the effects of glabridin(Gla)on intestinal flora and metabolites in mice were detected to explore the anti-tumor effect of Gla on colorectal cancer(CRC).Methods Twenty-four 6-week-old C57BL/6 male mice were randomly divided into blank group,model group,glabridin group and 5-fluorouracil group.The blank group and the model group were intraperitoneally injected with 100 μL normal saline,and the glabridin group was intraperitoneally injected with 100 μL glabridin solution(50 mg/kg).After 7 days of continuous administration,once every 2 days.The general condition of the mice was observed,and the samples were taken on the 22nd day.The tumor mass of each group was measured.HE staining was used to observe the changes of tumor tissue.IHC staining was performed to detect the expression of Ki-67,Cyclin D1,and Cleaved Caspase-3 in tumor tissues.16S rRNA was used to detect the changes of intestinal flora structure.Non-targeted metabolic detection of metabolite differences.Results Gla can significantly inhibit the growth of tumor in mice;HE staining showed that the density of tumor tissue in mice decreased after Gla intervention,the nucleus was small and lightly stained,and significant apoptotic areas were observed.IHC staining revealed that after Gla intervention,the expression of Ki-67 and Cyclin D1 was downregulated,while the expression of Cleaved Caspase-3 was upregulated(P<0.01)in tumor tissues.The results of 16S rRNA sequencing showed that Gla could change the structure of intestinal flora in mice,and the abundance of dominant bacteria was significantly changed.A total of 51 differential metabolites were screened by non-targeted metabolomics,and 9 potential pathways were obtained by enrichment analysis,including terpenoid skeleton biosynthesis,glutathione metabolism,peroxisome proliferator-activated receptor,HIF-1 signaling pathway,lysine degradation,GABAergic synapse,glycolysis/gluconeogenesis,cholesterol metabolism,nicotinic acid and nicotinamide metabolism.Conclusion Gla can inhibit the progression of CRC by improving the diversity of intestinal flora,regulating intestinal microecological balance and intestinal metabolites.