Omics Study of the Effects of Dachaihu Decoction on mRNA Expression and Non-targeted Metabolism in Ileal Tissue of Sepsis Rats
Objective:To investigate the therapeutic effect and mechanism of Dachaihu Decoction(DCHD)on intestinal injury in septic rats based on high-throughput RNA sequencing(RNA-seq)and ultra-performance liq-uid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometry(UPLC-QTOF-HRMS).Methods:Rat model of septic intestinal injury(SII)was established using the cecal ligation and punc-ture(CLP)method.90 rats were randomly divided into a sham operation group(n=10),a model group,and low,medium,and high dose groups of DCHD(n=20,each).The drugs were administered twice postoperatively,and sur-vival rates and murine severity scores of sepsis(mMSS)were observed after 24 hours.Hematoxylin and eosin(HE)staining was used to assess intestinal pathological changes.Immunofluorescence(IF)and real-time quantitative polymerase chain reaction(rt-PCR)were used to evaluate the expression of the tight junction protein Occludin in the intestine.Biochemical tests were performed for creatine kinase(CK),lactate dehydrogenase(LDH),CK-MB iso-enzyme,alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),blood urea nitro-gen(BUN),urea(UREA),and creatinine(Cr).Enzyme-linked immunosorbent assay(ELISA)was used to measure diamine oxidase(DAO)levels.RNA-seq and non-targeted metabolomics analysis were conducted using ileal tis-sues from the model group and the medium dose DCHD group.Results:The 24-hour survival rate in the model group was lower than that in the sham group,while there was no significant difference in survival rates between the various DCHD dose groups and the sham group.The medium dose of DCHD significantly reduced mMSS scores,improved intestinal injury,and increased Occludin expression.RNA-seq identified 205 differentially ex-pressed mRNAs,while metabolomics identified 968 differential metabolites.These differentially expressed mRNAs and metabolites were enriched in pathways related to lipid digestion and absorption,carbohydrate digestion and ab-sorption,protein digestion and absorption,and vitamin digestion and absorption,all showing positive regulation.Ad-ditionally,DCHD improved heart,liver,and kidney function in SII rats.Conclusion:DCHD can reduce mMSS scores,alleviate intestinal injury,and improve heart,liver,and kidney functions in septic rats.The mechanism may be associated with promoting the digestion and absorption of nutrients.
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