Study On Safety Evaluation Of Mongolian Medicine TuMuXiangSiWeiTang

Purpose:This study takes Tu Mu Xiang Si Wei Tang as an example drug,and explores the liver or kidney toxicity and toxicity mechanism of Tu Mu Xiang Si Wei Tang through subacute toxicity experiments,providing scientific basis for the safe and reasonable clinical application of Tu Mu Xiang Si Wei Tang.method:1.Subacute toxicity test:SD rats were randomly divided into high dose(16.0884g/kg),medium dose(2.6814g/kg),low dose(0.4469g/kg)groups of Sophora flavescens,high dose(18.2088g/kg),medium dose(3.0348g/kg),low dose(0.5058g/kg)groups of Tu Mu Xiang Si Wei Tang,Lacking sophora flavescens(18.2088g/kg)and normal control group.There were 8 groups in total,and rats in each treatment group were poisoned for 28 days.The biochemical indexes of serum,liver and kidney tissue were detected;Calculate organ index;Liver and kidney histopathological examination was performed.2.The effect of Mu Xiang Si Wei Tang on intestinal microbiota: Take the feces of rats in each group and sequence them using intestinal microbiota sequencing technology.To explore the effect of intestinal flora on hepatotoxicity of Sophora flavescens and Tuxiang Siwei Decoction.3.Based on the analysis and verification of the subacute toxicity mechanism of Mu Xiang Si Wei Tang by transcriptomics: take the liver of rats in each group,analyze the gene expression of liver tissue by transcriptomics analysis method,and verify the correlation between different genes and chemical components by molecular docking.result:1.The results of subacute toxicity test are as follows:(1)Effect of experimental drug on rat organ index:(1)Effect of Sophora flavescens group on liver organ index: compared with the normal group,the liver index of rats in the high and middle dose groups of Sophora flavescens increased significantly(p<0.05,p<0.01);The renal index of the high dose group of Sophora flavescens was significantly higher(p<0.05,p<0.01).(2)The effect of Tu Mu Xiang Si Wei Tang group on organ index: compared with the normal group,the kidney index of rats in the high-dose group of Tu Mu Xiang Si Wei Tang group increased significantly(p<0.05,p<0.01);The lung index of the rats in the middle dose group of Tu Mu Xiang Si Wei Tang group increased significantly(p<0.05).(3)The effect of the Kushen deficiency group on organ index:There was no significant difference compared to the Tu Mu Xiang Si Wei Tang.(2)Effects of experimental drugs on serum biochemical indexes of rats:(1)Effect of Sophora flavescens group on serum biochemical indexes of rats: Compared with the normal group,the AST content of Sophora flavescens was significantly higher(p<0.05).(2)The effect of the group of Tu Mu Xiang Si Wei Tang group on the serum biochemical indexes of rats: compared with the normal group,the ALT content of the high-dose group of Tu Mu Xiang Si Wei Tang group was significantly higher(p<0.05).(3)Effects of lack Sophora flavescens group on serum biochemical indexes of rats:There was no significant difference between the group of lack Sophora flavescens and the group of Tu Mu Xiang Si Wei Tang group.(2)Effects of experimental drugs on pathological changes of rats:(1)Effect of Sophora flavescens on pathological changes of liver and kidney in rats:The Sophora flavescens group showed varying degrees of abnormalities in both the liver and kidneys at high doses.(2)The effect of the group of Civil Fragrance Siwei Decoction on the pathological changes of liver and kidney in rats:There were varying degrees of abnormalities in the liver and kidneys in the group of Tu Mu Xiang Si Wei Tang and in the high-dose group.(3)Effect of lack Sophora flavescens group on the pathological changes of liver and kidney: Compared with the group of Tu Mu Xiang Si Wei Tang group.there was no significant difference in lack Sophora flavescens group.2.The results of the effect of Tu Mu Xiang Si Wei Tang on intestinal flora are as follows:(1)The Venn diagram analysis results of the sample show that: The Venn diagram analysis results of samples showed that:The specific OTUs of the normal group,the Sophora flavescens group,the Tu Mu Xiang Si Wei Tang group and lack Sophora flavescens group were 2059,2862,1952 and 1798.(2)The results of significant difference analysis of sample flora show that:Compared with the normal control group,the significantly different bacterial groups of Sophora flavescens group is composed of 36 flora including Clostridia、Clostridiales 、 Ruminococcaceae 、 Proteobacteria 、 Deltaproteobacteria 、Desulfovibrionaceae 、 Desulfovibrionales 、 Lachnospiraceae 、 Gemmataceae 、Haliangiaceae 、 Epsilonproteobacteria 、 Helicobacteraceae 、 Campylobacterales 、Odoribacteraceae、Microbacteriaceae、Dehalobacteriaceae、Anaeroplasmataceae、Mollicutes 、 Anaeroplasmatales 、 Tenericutes 、 Caulobacterales 、Caulobacteraceae 、 Rhizobiaceae 、 Peptococcaceae 、 Veillonellaceae 、Christensenellaceae 、 Barnesiellaceae 、 Deferribacteraceae 、 Deferribacteres 、Mycobacteriaceae、Nitrospiraceae、Syntrophobacteraceaeo、Syntrophobacterales、Rhodospirillaceae 、 Acidimicrobiia.etc.The significantly different microflora in the Tu Mu Xiang Si Wei Tang are composed of 19 microflora,including Epsilonproteobacteria,Helicoproteobacteraceae,Campylobacteres,Bacteroidaceae,Christensenellacea,Mogibactriacea,Turcicibecteraceae,Desulfovibrionacea,Deltaproteobacteria,Proteobacteria,Barnesiellacea,Deferriberaceae,Firmicutes,Lachnospiracea,Clostridia,etc.The significantly different flora of lack Sophora flavescens group are Paraacteroides,Porphyromonadaceae,Prevotella,Prevotella,Bacteroidales,Bacteroidia,Bacteroidetes,rc4＿4.Peptococcaceae,Bilphila,Dosulfovibrionaceau,Desufovibrionales,Deltaprotenhacleria)and other 11 bacterial groups.(3)Correlation analysis results show that:AST is negatively correlated with Helicobactercea,Gemmataceae,Desulfovibrionacea,Dehalobactriacea,Peptococcaceae,Caulobacteracea,Deferribaceracea,Odoribacteracea and other microflora,and positively correlated with Erysipelotrichaceae,Bifidobacteriaceae,Lactobacillaceae,Clostridiacae,Coriobacteriacese and other microflora.ALT is positively correlated with the flora of Ellin6075,Verrucomicrosbiaceae,Metrolocaceae,Christensenellaceae,and negatively correlated with the flora of Burkholderiaceae.3.Based on transcriptomics,the analysis and validation results of subacute toxicity mechanism of Tu Mu Xiang Si Wei Tang are as follows:(1)Differentially expressed genes: Compared with the normal control group,the differentially expressed genes in Sophora flavescens group were down-regulated by Cyp2b1,Grin2 c,Slc16a5,Gpat3,Akr1b7,Cyp2b2,Aldh1a7,Sult2a2,etc.,and up-regulated by Obp3,Mst1 r,Pnpla5,Mup4,Dhrs7l1,RGD1559960,Kcnj15,Scd,Lgsn,etc;The down-regulated differentially expressed genes in the Tu Mu Xiang Si Wei Tang were Cyp2b1,Grin2 c,Akr1b7,Cyp2b2,Sult2a2,Slc16a5,Acot1,Abcc3,Aldh1a7,etc.The up-regulated ones are Obp3,Mst1 r,Stac3,Fras1,Pnpla5,Kcnj15,Mup4,Dhrs7l1,etc.Compared with Tu Mu Xiang Si Wei Tang,the differentially expressed genes in lack Sophora flavescens group were down-regulated by Obp3,Acot1,Pbk,Slc22a12,Lexm,Rmi2,Cdca3,Hmmr,Top2 a,Wfdc6c,Rad51,Cdk1,etc.,and up-regulated by Pnpla3,Ppp1r3 g,Noct,Pkdrej,edil3,etc.(2)GO and KEGG enrichment analysis results show that:It is related to bile secretion,cytochrome P450 metabolism of exogenous substances,arachidonic acid metabolism and glutathione metabolism and other enrichment pathways.(3)The results of molecular docking of rat transcriptomic differential genes and chemical components showed that the differential genes successfully docking with five chemical components including matrine,oxymatrine,sophoridine,sophocarpine and oxysophocarpine were Kcnj15,Sult2a2,Cyp2b2,Akr1b7 and Aldh1a7.Conclusion:(1)Tu Mu Xiang Si Wei Tang has no toxicity within 28 days of clinical dosage;but36 times the clinical dose has subacute liver toxicity;Hepatotoxicity is mainly related to sophora flavescens.(2)The significant difference in the bacterial flora of the Tu Mu Xiang Si Wei Tang is related to the disorder of bacteria such as Desulfovibrio,Clostridium and Proteus.(3)After molecular docking of differentially expressed genes related to Tu Mu Xiang Si Wei Tang,it was found that Kcnj15,Sult2a2,Cyp2b2,Akr1b7,and Aldh1a7 had higher docking scores with toxic components such as matrine,oxymatrine,sophoridine,sophocarpine,and oxysophocarpine in Sophora flavescens.It is preliminarily believed that Kcnj15,Sult2a2,Cyp2b2,Akr1b7,Aldh1a7 and other differential genes are the molecular mechanism of hepatotoxicity caused by Tu Mu Xiang Si Wei Tang.