Study On Multi-group Toxicological Mechanism Of Compound Nano-realgar

Obigctive:His project intends to study the toxicology of compound nano-realgar prescription and single drug in mice.Through the quantitative macrogenomic study of intestinal flora,the metabolic group of peripheral blood,the transcription group of peripheral blood and main organs,the overall toxicity mechanism of compound nano-realgar and its single drug in mice is revealed from the overall point of view,and hopes to provide the theoretical basis for toxicological prevention for patients in the future.Method:SPF male C57BL/6 mice were randomly divided into four groups.300 mg fecal samples was collected as a control before administration.Compound nano-realgar,indigo naturalis,nano-realgar and 5% sodium carboxymethyl cellulose were orally administered for five weeks,once a day,and 300 mg feces of mice were collected weekly after administration.After 5 weeks,the anesthetized mice were collected peripheral blood and peripheral blood serum by eyeball extraction.The heart,liver and kidney of mice were dissected and collected to prevent RNA degradation.Some liver,kidney and intestine tissues were fixed with formaldehyde for pathological sections,and the rest tissues were stored in cold storage.Total DNA was extracted from the collected fecal samples of mice and sequenced at a depth of more than 50,000 stitched microbial gene sequences per sample.Total RNA was extracted from peripheral blood for sequencing analysis.The depth of sequencing was 6G clean data for each sample.Total RNA was extracted from heart,liver and kidney tissue samples,and the sequencing method was the same as that of peripheral blood RNA.Peripheral blood serum samples from each group were pretreated before the experiment.The metabolomics of peripheral blood serum was analyzed by UPLC-Q/TOF-MS high-resolution liquid-mass spectrometry.Result:1.Compared with the tissue samples taken from the control group,the liver of Indigo Naturalis group was slightly damaged,and no pathological changes were observed under light microscopy in other tissue samples.2.The expression of m RNAs in multiple tissues and organs of the transcriptome was abnormal,and 1943 differential genes were identified,of which 625 were up-regulated and 1251 were down-regulated.GO enrichment analysis of compound nano-realgar blood group showed that differential gene m RNAs mainly activated the negative regulation of angiogenesis,cell response to acidic chemicals and other functions;inhibited red blood cell development,autophagy and other functions.KEGG enrichment analysis showed that differentially expressed genes mainly activated osteoclast differentiation pathway and inhibited ubiquitin-mediated protein hydrolysis pathway.GO enrichment analysis of compound nano-realgar heart tissue showed that differential gene m RNAs mainly activated cell-matrix adhesion,single cell-cell adhesion and other functions;inhibited heart conduction,regulated heart rate,heat response and other functions.KEGG enrichment analysis showed that differentially expressed genes mainly activated platelet activation and PI3K-AKT pathways,and inhibited protein processing pathways in endoplasmic reticulum.GO enrichment analysis of compound nano-realgar kidney tissues showed that differential gene m RNAs mainly activated metabolic processes,lipid storage and other functions;inhibited the negative regulation of RNA polymerase II promoter transcription,hematopoiesis and other functions.KEGG enrichment analysis showed that differentially expressed genes mainly activated retinol and arachidonic acid metabolism pathways,and inhibited meiosis pathways of oocyte.GO enrichment analysis of compound nano-realgar liver tissue showed that differential gene Mrs mainly activated the cell response to lipid,positive regulation of lipid metabolism,inhibition of long-chain fatty acid metabolism,unsaturated fatty acid biosynthesis and other functions.KEGG enrichment analysis showed that differentially expressed genes mainly activated complement and coagulation cascade pathway,and inhibited PPAR signaling pathway.3.Serum metabonomics analysis showed that in the OPLS-DA model,the samples of control group,Indigo Naturalis group,nano-realgar group and compound nano-realgar group could be well distinguished,indicating that the metabolites characteristics of the three groups were significantly different from those of the control group.KEGG analysis showed that there were 16 metabolic pathways in the compound nanorealgar group compared with the control group,which were significantly enriched in the biosynthesis of unsaturated fatty acids,aminoacyl-t RNA biosynthesis,ABC transporter,purine metabolism,metabolic pathway and tyrosine metabolism signaling pathways.4.16 S r RNA gene and macrogenome sequencing showed that the diversity,homogeneity and diversity of bacterial flora in each group changed significantly compared with the control group,but there was no correlation with the time of administration.Compared with the control group,the KEGG pathways involved in the differential metabolites of compound nano-realgar were mainly concentrated in the biosynthesis of sstilbenoid,diarylheptanoid and gingerol,the metabolism of xenobiotics by cytochrome P450,nicotinate and nicotinamide metabolism,MAPK signaling pathway,glutathione metabolism,pentose phosphate pathway and drug metabolism-cytochrome P450 seven signaling pathways.Conclusion:1.After the administration of compound nano-realgar,he abnormal expression of m RNAs and serum metabolites in multiple tissues and organs and the difference of intestinal flora in animals changed significantly.These differential molecules showed various biological functions and participated in various biological reactions and signal transduction pathways.2.Screening of differential molecules has important guiding significance for finding the toxicological mechanism and pharmacological mechanism of compound nano-realgar.