Analysis Of The Rule Of NAFLD Prescription And Investigation Of TCM Syndrome Characteristics And Study On The Mechanism Of Bulbus Allii Fistulosi Extract In The Treatment Of NAFLD Rats

This paper discusses the cognition of non-alcoholic fatty liver disease by traditional Chinese medicine through literature analysis,clinical investigation,network pharmacology and animal experiment.Part one.Analysis on prescription rules of treating NAFLD based on traditional Chinese medicine inheritance auxiliary systemsObjective Through using traditional Chinese medicine inheritance auxiliary system to analyze the prescription rules of NAFLD.Methods We collect documents in CNKI and Wan Fang Data used for treating NAFLD.By using Traditional Chinese Medicine Inheritance Platform Management system,we established formula databases based on association rules,cluster analysis method and complex system entropy cluster.Results A total of 320 prescriptions were included,involving 216 kinds of herbs,6 kinds of main syndrome types and 9 kinds of common treatments.The frequency of drug use,four qi and five flavors,meridian return and the association and compatibility of drugs were summarized.Conclusion Data mining is used to conduct correlation analysis on the prescriptions of NAFLD,and it is concluded that the prescriptions are mainly aimed at eliminating phlegm and dispersing constipation,promoting blood circulation and removing blood stasis,and the rules of the prescriptions are further summarized to provide some reference for clinical diagnosis and treatment.Part two.Investigation on the characteristics of TCM syndromes of non-alcoholic fatty liver diseaseObjective To investigate the TCM syndromes,syndromes distribution and characteristics of NAFLD by collecting clinical data.Methods Ninety-eight patients with NAFLD were included in the study.On the basis of TCM dialectics and related guidelines,patients Dialectical classification will be included.Symptoms,general conditions,clinical examination indexes and scale scores of each patient were collected,so as to summarize and discuss the relationship between TCM syndrome types and objectification indexes.Results Among the 98 patients,the incidence of male was higher than female.The morbidity of both sexes tended to be younger;Abnormal elevation of blood lipid(40.8%),hypertension(41.8%)and abnormal glucose tolerance(31.6%)were the most common risk factors.There were 31 cases of liver depression and spleen deficiency syndrome,28 cases of dampness-heat accumulation syndrome,19 cases of phlegm dampness internal obstruction syndrome,13 cases of phlegm and blood stasis interconnection syndrome,and 7 cases of spleen-kidney deficiency syndrome.The 10 syndromes with the most frequent occurrence were limb weakness(66),fullness of ribs(61),loss of appetite(54),epigastric puffiness(48),unpleasant stool(36),dry mouth(31),difficulty sleeping(31),nausea(30),greasy mouth(29),and hypochondria(28).ALT increased compared with other syndromes of liver stagnation and spleen deficiency and phlegm dampness internal obstruction.The levels of triglyceride and total cholesterol were the highest in the syndrome of mutual association of phlegm and blood stasis.The fasting blood glucose and water content of dampness-heat syndromes were higher than other syndromes.In terms of PRO scale score,the syndrome of mutual association of phlegm and blood stasis had the highest score in the physiological field,the syndrome of liver stagnation and spleen deficiency had the highest score in the psychological field,and the syndrome of mutual association of phlegm and blood stasis had the highest total score.Conclusion The incidence of NAFLD is high in males and tends to be younger.Abnormal hyperlipidemia,hypertension and abnormal glucose tolerance were the most common risk factors.Liver stagnation and spleen deficiency were the most common syndromes.Extremities weakness,hypochondriac distension full,inappetence,epigastric puffiness and other syndromes are common witnesses;There were differences in ALT,triglyceride,total cholesterol,fasting blood glucose and PRO scale scores among different syndromes,but these differences were further verified by increasing the sample size.Part three.Study on the mechanism of treatment of nonalcoholic fatty liver disease with Bulbus Allii Fistulosi extract based on network pharmacologyObjective To investigate the potential targets of Bulbus Allii Fistulosi extract in treatment of NAFLD based on network pharmacology.Methods The Traditional Chinese Medicine System Platform(TCMSP)database was used to find out active components and potential targets of Bulbus Allii Fistulosi extract.Then the potential target information of NAFLD was retrieved by Genecards databases.Perl language software was used for preprocessing,and the R language software,Cytoscape software and String network data platform were used data analysis and picture production.Results There were 17 common gene targets of ?-sitosterol and NAFLD.By GO function enrichment analysis,it was found that the functions of common gene targets involved nuclear receptor activity and transcription factor activity.KEGG pathway enrichment analysis showed that common gene targets involved fluid shear stress and atherosclerosis signaling pathway,TNF signaling pathway,apoptosis signaling pathway,IL-17 signaling pathway,diabetes complications signaling pathway,adipokine signaling pathway,insulin resistance signaling pathway,NF-?B signaling pathway,etc.Conclusion Based on the results of network pharmacological analysis and literature review,PPAR? was predicted to be an important target of scallion extract in the treatment of NAFLD.Part four.Experimental study on the improvement of insulin resistance and macrophage polarization in NAFLD rats by Bulbus Allii Fistulosi extract based on PPAR?Objective To investigate the mechanism of improving insulin resistance and macrophage polarization in NAFLD rats by Bulbus Allii Fistulosi extract.Methods 36 male rats were modeled by high fat diet and randomly divided into normal group,model group,Bulbus Allii Fistulosi extract(BAF)group,BAF plus inhibitor group,pioglitazone group and inhibitor group.The body weight and liver wet weight of the rats were measured at the end of the experiment.Serum samples were collected to detect the levels of TC,TG,AST,ALT,FBG,FINS,HOMA-IR.Skeletal muscle specimens were collected for HE staining,and GLUT4 and CD68 protein expressions were detected by immunofluorescence staining.The protein expressions of IRS-1,PI3 K and GLUT4 were detected by Western blot.The expressions of F4/80,i NOS,TNF?,Arg-1 and IL-10 were detected by realtime fluorescence PCR.Liver tissues were collected for HE staining and oil red O staining.The protein expressions of CD68,CD163,TNF?,IL-10,TLR4 and My D88 were detected by immunofluorescence staining.The protein expressions of PPAR?,CD68,TLR4,NF?B and My D88 were detected by Western blot.The expressions of IRS-1,IRS-2,SREBP-1c,PEPCK,G6 Pase,F4/80,MCP-1,i NOS,Arg-1,TNF? and IL-10 were detected by real-time fluorescence PCR.Results Bulbus Allii Fistulosi extract and pioglitazone could control the body weight of NAFLD,and reduce the levels of TC,TG,AST,ALT,FBG,FINS,HOMA-IR.Increase the GLUT4 and CD68 immunofluorescence intensity of skeletal muscle;Upregulate the protein expression of IRS-1,PI3 K and GLUT4 in skeletal muscle;The expressions of Arg-1 and IL-10 in skeletal muscle were upregulated,and the expressions of F4/80,i NOS and TNF? were down-regulated.The fluorescence intensity of CD68,TNF?,TLR4 and My D88 protein in liver tissue was decreased,and the fluorescence intensity of CD163 and IL-10 protein was increased.The expression of PPAR? protein in hepatic tissue was upregulated,and the expression of CD68,TLR4,NF?B,and My D88 protein were down-regulated.The expressions of ARG-1,IL-10 and IRS-2 in hepatic tissue were up-regulated,and the expressions of IRS-1,SREBP-1c,PEPCK,G6 Pase,F4/80,MCP-1,i NOS and TNF? were down-regulated.There was no statistical difference in the above indicators between the BAF plus inhibitor group and the model group.Conclusion Bulbus Allii Fistulosi extract can improve insulin resistance of NAFLD,induce macrophage transformation from M1 type to M2 type,and reduce immune inflammatory response.At the same time,PPAR? is one of the important molecular targets for the therapeutic effect of Bulbus Allii Fistulosi extract.