Study On The Effect Mechanism Of Traditional Chinese Medicine Compound Prescription GC In The Treatment Of Experimental Nonalcoholic Steatohepatitis Based On Liver Immune Regulation

Objective:Based on liver immune regulation,the mechanism of action of experimental NASH in the treatment of experimental NASH by the combination of Geniposide and Chlorogenic acid.Methods:Induced experimental steatohepatitis(NASH)model in mice by high trans fat high cholesterol high bile salt feed compound with high sugar drinking water for 20 weeks.Dosing started at week 20,the experimental mice were randomly divided into normal group(Control),high fat high glucose group(HFHC)and medication group(GC),8 in each group.After 5 weeks of administration,the materials were harvested,and observed:(1)pharmacodynamic indicators,including:(1)liver tissue triglyceride content(TG);(2)the activity of serum alanine aminotransferase(ALT),serum aspartate aminotransferase(AST);(3)serum low density lipid protein cholesterol(LDL-C),serum total cholesterol(Tch);(4)fasting blood glucose(GLU),fasting insulin(Insulin)and insulin resistance index(HOMA-IR);(5)liver histopathology(HE,Sirius red);(6)Liver tissue type I collagen(COL-?),? smooth muscle actin(?-SMA);(2)liver immune related indicators,including:(1)flow cytometry analysis of liver immune cells CD3+T cells,CD4+T cells,CD8+T cells,B cells,NK cells,NKT cells,DC cells;(2)Liver tissue macrophage marker F4/80 and neutrophil marker MPO immunohistochemical staining;(3)liver tissue interleukin-like pro-inflammatory and anti-inflammatory factors,chemokines and Colony stimulating factor protein content.Results(1)Pharmacodynamic results:(1)GC significantly decreased the TG content in liver tissue of NASH mice(P<0.05);(2)GC significantly decreased serum ALT and AST activity(P<0.01);(3)GC significantly decreased serum LDL-C and serum Tch levels(P<0.01);(4)GC significantly decreased serum fasting INS level(P<0.05);(5)histologically GC significantly improved liver tissue steatosis,inflammation and balloonlike degeneration,reduced the positive staining area of liver Sirius red;(6)GC reduced the expression of COL-I and ?-SMA in liver tissue;(2)Liver immunity related results:(1)GC significantly down-regulated the proportion of liver tissue NKT cells,CD3+T and CD4+T cells(P<0.05);(2)GC reduced the protein expression of liver tissue macrophage surface marker F4/80 and neutrophil marker MPO;(3)GC significantly down-regulated the m RNA level of Th1 nuclear transcription factor T-bet(P<0.05);(4)Liver tissue inflammatory factor results: In terms of interleukin-like inflammatory factors,GC significantly reduced pro-inflammatory IL-1?,IL-1?,IL-2,IL-6,IL-12 protein levels(P < 0.01),reduced anti-inflammatory IL-10 protein levels(P < 0.05);in terms of tumor necrosis factor and interferon-like inflammation,GC significantly reduced the protein levels of TNF? and interferon gamma(IFN?)(P<0.05);in terms of chemokines,GC significantly decreased the protein levels of monocyte chemoattractant protein 1(MCP-1),CXC motif chemokine 1(CXCL-1),CC motif chemokine 11(CCL-11)(P<0.01 or P<0.05);In terms of colony stimulation factors,GC significantly reduced the protein levels of granulocyte-macrophage colony-stimulating factor(GM-CSF)and granulocyte colony-stimulating factor(G-CSF)(P<0.05).Conclusion(1)Traditional Chinese medicine compound of GC significantly improved liver lipid deposition,inflammation and fibrosis in NASH mice;(2)GC significantly reduced the proportion of liver NKT cells,CD3+ T cells and CD4+ T cells in NASH mice,inhibit the expression of macrophages,and regulated liver immunity.