Study On The Action Mechanism Of Xuefu Zhuyu Decoction With Different Compatibility Forms Of Meridian-Introducing Drugs On NAFLD

Objective:By establishing a C57BL/6J Apoe^-/-mouse non-alcoholic fatty liver disease（NAFLD）animal model,and giving different forms of compatible preventive intervention with Xuefu Zhuyu Decoction to evaluate the effect of Xuefu Zhuyu Decoction on the NAFLD model.The effect of neutrophil infiltration in mouse liver tissues and exploring its mechanism of action provide laboratory evidence for Xuefu Zhuyu Decoction to treat NAFLD.Method:36 C57BL/6J Apoe^-/-mice were selected,and 6 were randomly selected as the Control group.The remaining mice were injected with high-fat diet combined with subclinical dose of LPS（4ng/kg）intraperitoneally to establish NAFLD model,and the model animals were randomly selected divided into Xuefu Zhuyu decoction group（XFZY group）,Xuefu Zhuyu decoction minus achyranthes bidentata group（XFZY-N group）,Xuefu Zhuyu decoction minus Platycodon grandiflorum group（XFZY-J group）,and Xuefu Zhuyu decoction minus Achyranthes bidentata and Platycodon grandiflorum group（XFZY-NJ）,model group（Model group）were given an equal volume of saline intragastric intervention for 4 weeks.HE staining and oil red O staining were used to observe the pathological changes of liver tissue,immunofluorescence method was used to detect the number of Ly6G⁺cells in mouse liver tissue,flow cytometry was used to detect the proportion of detect the concentration of KC and MIP-2 in mouse plasma and the concentration of MIP-2 and MPO in liver tissue.Western blot was used to detect the protein expression of p38MAPK,phospho-p38MAPK,MKP7,Caspase-3,and PARP in mouse liver tissue.The amount of Bcl-2,Bax,Fas,Fas L gene expression in mouse liver tissue was detected by RT-PCR.Results:Compared with the Model group,the proportion of CD14⁺CD11b⁺Ly6G⁺cells in the peripheral blood of mice in the XFZY,XFZY-N,and XFZY-J groups all decreased（P<0.05）.Compared with the XFZY group,in the XFZY-NJ group,the percentage of CD14⁺CD11b⁺Ly6G⁺cells increased（P<0.05）;Compared with the Model group,the KC content in the peripheral blood of mice in the XFZY group,XFZY-N group,XFZY-J group,and XFZY-NJ group decreased group（P<0.05）;And compared with Model group,XFZY group,XFZY-N group,XFZY-J group had lower levels of MIP-2 in peripheral blood（P<0.05）.Compared with XFZY group and XFZY group,XFZY-NJ group had MIP-2 content Increased（P<0.05）;Compared with the Model group and XFZY-NJ group,the XFZY,XFZY-N,and XFZY-J groups all have different degrees of reduction in liver steatosis and inflammatory infiltration.The XFZY group is the most obvious;Compared with the Model group,XFZY-N group,XFZY-J group and XFZY-NJ group,the degree of neutrophil infiltration in the liver tissue of the XFZY group was significantly improved;Compared with the Model group,the content of MIP-2 in the liver tissues of mice in the XFZY and XFZY-N groups decreased（P<0.05）,and the XFZY group was lower than the XFZY-NJ group（P<0.05）;Compared with the XFZY group,the XFZY-J group mice MPO content in liver tissue increased（P<0.05）;Compared with the Model group,the relative expression of p38MAPK and phospho-p38MAPK in the liver tissue of mice in the XFZY,XFZY-N,and XFZY-J groups decreased（P<0.05）;Compared with the XFZY group,the relative expression of MKP7in the Model group and the XFZY-NJ group increased（P<0.05）;Compared with the Model group,the relative expression of PARP in the XFZY group and the XFZY-N group decreased（P<0.05）;Compared with the Model group,the relative expression of Caspase-3 in the XFZY group and the XFZY-J group was reduced（P<0.05）.Compared with the XFZY group,the Caspase-3 in the XFZY-N group and the XFZY-NJ group were relatively expression level increased（P<0.05）.Compared with the XFZY-J group,the relative expression of Caspase-3 in the XFZY-NJ group increased（P<0.05）;Compared with the Model group and the XFZY-NJ group,the relative expression of Bax in the XFZY group,the expression decreased（P<0.05）,the relative expression of Bcl-2 in the XFZY group increased（P<0.05）;Compared with the Model group,the relative expression of Fas in the XFZY,XFZY-N and XFZY-J groups decreased（P<0.05）,the relative expression of Fas L in the XFZY group,XFZY-N group and XFZY-J group decreased（P<0.0 5）;Compared with the XFZY group and the XFZY-J group,the relative expression of Fas L in the XFZY-NJ group increased（P<0.05）.Conclusion:1.Xuefu Zhuyu Decoction can prevent liver tissue steatosis and liver inflammation in NAFLD model animals,improve general symptoms of mice,reduce the relative expression of p38MAKP,inhibit LPS/p38MAPK signaling pathway,inhibit neutrophil activation,and reduce the tendency.The concentration of chemical factors KC and MIP-2 prevents the recruitment of activated neutrophils to the inflammation site,reduces the expression of pro-apoptotic protein Caspase-3 and pro-apoptotic genes Bax and Fas,and promotes the expression of apoptosis-inhibiting gene Bcl-2Expression;2.Compatible with the meridian drugs Achyranthes bidentata and Platycodon grandiflorum in the Xuefu Zhuyu Decoction,have a certain synergistic effect in inhibiting the LPS/p38MAPK signaling pathway,reducing the release of inflammatory factors and inhibiting cell apoptosis;3.Yin Jing The drug Achyranthes bidentata significantly promotes the expression of MKP7 protein and can inhibit the expression of Caspase-3 protein.