Study On The Effect Of The Uyghur Medicine Smiz Outi On The Regulation Of S100A8/A9 Expression And Inhibition Of CCl₄ Acute Liver Injury

Background:Various liver diseases caused by liver injury seriously threaten people's health.The continuous oxidative stress and inflammation of the liver play an important role in the occurrence and development of liver injury.Inflammation of the liver has a dual role in the liver and is essential for maintaining tissue health.Once the inflammatory response is excessive or unregulated,it will become a key driver of pathological inflammation and tissue damage,worsening the course of liver injury.The holistic view of traditional medicine and syndrome differentiation have unique advantages in the comprehensive treatment of liver injury.Traditional Chinese medicine has played a huge role in the treatment of acute liver injury due to its multi-target and multi-component effects.Objective:In this project,a model of acute liver injury in mice induced by carbon tetrachloride(CCl4)was used to investigate the role of Portulaca oleracea L(PO)in preventing and treating acute liver injury,and to explore the molecular mechanism of the liver protection effect of PO,The PO component with the lowest toxicity and the best pharmacological activity with S100A8/S100A9 as the target is clearly defined.Methods:Taking the acute liver injury model of mice induced by CC14 as the research object,60 SPF male Kunming mice were randomly divided into normal group,model group,silybin group(200 mg·kg-1)and high(2 mg·kg-1),medium(1 mg·kg-1),low(0.5 mg·kg-1)dose PO group.Five days after intragastric administration,in order to establish a mouse model of acute liver injury,in addition to the normal group of mice,the mice in each group were intraperitoneally injected with 0.2%CCl4 peanut oil solution 10 mg·kg-1 to induce liver injury.After 23 hours of modeling,serum and liver tissue were collected.Automatic biochemical analysis method was used to detect serum liver function indexes such as ALT,AST and TBIL in mice;hematoxylin-eosin(HE)combined staining method was used to stain liver pathological sections to observe liver pathological changes;transcriptome sequencing technology(RNA-seq)analysis of liver differential genes and GO function,KEGG enrichment,and real-time fluorescence quantitative reaction(Real-time PCR)to detect gene expression of Hsp90,C/EBP?,CCL2,S100A8 and S100A9,western blot to detect S100A9 protein expression level,ELISA to detect S100A8,S100A9 and S100A8/A9 concentration.Molecular docking,drug similarity,ADMET and other methods were used.to identify the anti-inflammatory and immunomodulatory chemical components in PO.Results:1.Study on the liver protection effect of POCompared with the normal group,the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),lactate dehydrogenase(LDH),and total bilirubin(TBIL)in the model group were significantly increased(P<0.01);compared with the model group,PO significantly reduced the levels of ALT,AST and LDH in acute liver injury mice(P<0.05,P<0.01).PO significantly improved pathological phenomena such as hepatocellular degeneration,swelling,necrosis,and inflammatory cell infiltration.2.Study on the molecular mechanism of the protective effect of PO on carbon tetrachloride-induced acute liver injury in miceCompared with the normal group,the Hsp90,C/EBP?,CCL2,S100A8 and S100A9 gene expression levels in the liver of the model group were significantly increased(P<0.05,P<0.01),and the protein levels of serum S100A8 and liver S100A9 were significantly increased(P<0.05,P<0.01);compared with the model group,the gene expression of Hsp90,C/EBPP,CCL2,S100A8 and S100A9 in the liver of mice of PO increased dose group was significantly reduced(P<0.05,P<0.01),Serum S100A8,S100A8/A9 and liver S100A9 protein levels were significantly reduced(P<0.05,P<0.01).3.In silico Study of the Anti-Inflammatory Activity of PO IngredientsFrom the scifinder and PubChem databases,70 PO compounds have been collected.There are 16 PO compounds strongly combined with S100A8/A9.Among them,portulacerebroside D,N-trans-hibiscusamide,oleraciamide B can be combined with S100A8/A9 forms hydrogen bonds.The drug-like properties of oleraciamide B,N-cis-hibiscusamide,and portulene are the best,followed by N-trans-hibiscusamide,racemic oleracein E,cycloartenol and lupeol.Conclusion:1.PO may affect the expansion of inflammatory response and recruitment of inflammatory cells by regulating the mRNA expression of C/EBP?,S100A8,S100A9 and CCL2 in mouse liver and serum S100A8?S100A8/A9 protein level,thereby reducing the acute liver induced by CCl4 damage.2.The oleraciamide B and N-trans-hibiscusamide in PO have the lowest toxicity and the best pharmacological activity against the designated target S100A8/A9.These compounds are expected to show great efficacy in combating inflammatory diseases.