Study On The Mitigating Effect And Mechanism Of Notoginseng R1 On Liver Fibrosis In Mice

Liver fibrosis is a pathological repair response to various liver injuries,and is a common stage of chronic liver disease progression to cirrhosis.The main feature of its pathogenesis is the abnormal secretion and excessive deposition of extracellular matrix after activation of liver muscle fibroblasts,leading to fibrosis in liver tissue.Studies have shown that liver fibrosis is a potentially reversible dynamic process,and if the fibrotic lesions are not promptly alleviated or eliminated at the initial stage of the disease,it may develop into irreversible cirrhosis.Therefore,the early reversal of liver fibrosis is crucial,but currently available clinical treatment methods are limited.Notoginseng R1 is a rich and unique tetracyclic triterpenoid saponin in Panax notoginseng.Modern pharmacological studies have found that it has extensive pharmacological activities.In previous studies,we found that Notoginseng R1 may have the potential to alleviate liver fibrosis in mice,but its dose,efficacy and pharmacological mechanism are not clear,Therefore,this study intends to further explore the effect and mechanism of notoginseng R1 in improving liver fibrosis in mice.The main research contents include the following three aspects:1.Notoginseng R1 effectively alleviates the pathological level of liver fibrosis in mice84 C57BL/6 mice were randomly divided into 7 groups,with 12 mice in each group: blank group(Control group),notoginseng R1 group(NG-R1 group),model group(Model group),positive drug control group(UDCA group),notoginseng R1 low dose administration model group(NG-R1-L group),Notoginseng R1 medium dose administration model group(NG-R1-M group),and Notoginseng R1 high dose administration model group(NG-R1-H group).After successfully constructing a mouse model of liver fibrosis,the experimental mice were continuously administered for 14 days.The experimental mice were killed and tissue samples such as blood,liver,ileum,and spleen were collected.Detection of ALT,AST,AKP γ-GT content;Detection of the content of HYP in the liver;The pathological changes of liver tissue were observed by H&E and Masson staining;Analysis of liver tissue in mice with liver fibrosis using protein immunoblotting and real-time fluorescence quantitative PCR α-SMA/TGF-β 1/Col-1/IL-6/TNF-α Protein and gene expression levels.The results showed that Notoginseng R1 could slow down the weight loss,restore the liver index and spleen index,and improve the vitality of mice with liver fibrosis.Notoginseng R1 reduced ALT,AST,AKP γ-The content of GT can significantly improve the level of liver function,effectively alleviate the damage of liver morphology and liver tissue structure in liver fibrosis mice,reduce inflammatory cell infiltration and collagen deposition in the liver,and reduce inflammatory factors(IL-6,TNF-α)And fibrosis factor(TGF-β 1、 α-The expression of SMA and Collagen Ⅰ can reduce the content of HYP and improve liver fibrosis.2.Notoginseng R1 can improve liver fibrosis in mice by reducing liver bile acid synthesis and promoting liver bile acid excretion.Detect the content of TBA,TBIL,and DBIL in serum;The expression levels of FGFR4/m TOR/TFEB/CYP7A1 gene and protein in liver tissue of mice with liver fibrosis were detected by real-time fluorescence quantitative PCR and Western blot analysis;Protein immunoblotting and real-time fluorescence quantitative PCR were used to analyze the expression level of FXR/FGF15 protein and gene in the mouse ileum;The expression level of NTCP/BSEP/OSTEP liver gene in mouse liver tissue was detected by real-time fluorescence quantitative PCR.The results showed that Notoginseng R1 could reduce the concentration of bile acid in the blood of mice with liver fibrosis.The mechanism was that Notoginseng R1 inhibited the expression of bile acid synthase CYP7A1 in the liver through FXR-FGF15-TFEB,thereby reducing the synthesis of bile acid,reducing the concentration of bile acid in mice,thereby slowing down liver fibrosis induced by bile duct ligation in mice.In addition,Notoginseng R1 inhibits the expression of NTCP,activates the liver expression of BSEP and OSTP mice,reduces the entry of bile acids from the blood into the liver,and promotes the elimination of bile acids from the liver,thereby reducing the accumulation of bile acids in the liver and maintaining the homeostasis of bile acids in the liver.3.3.Effect of Notoginseng R1 on intestinal microflora changes in mice with liver fibrosisH&E staining was used to observe the histopathological changes of the ileum;Analysis of IL-6/TNF in the ileum of mice with liver fibrosis using real-time fluorescence quantitative PCR and Western blotting-α Expression levels of genes and proteins;Changes in intestinal microorganisms in mouse feces were analyzed using16 S r RNA sequencing.The results showed that Notoginseng R1 reduced the inflammatory factor(TNF)in the ileum-α、 The expression of IL-6)can improve the invasion of ileitis,alleviate intestinal damage in mice with bile duct ligation,and help restore the normal structure of the ileum.In addition,Notoginseng R1 can effectively improve the disorder and decrease in diversity of intestinal flora in mice caused by bile duct ligation,improve the species uniformity,richness,and diversity of intestinal flora,increase the beneficial flora of intestinal microorganisms,reduce the pathogenic flora,and alleviate liver fibrosis caused by bile duct ligation by improving the flora changes of intestinal microorganisms.