| High-fat,high-calorie diet has been considered to be the leading cause of NAFLD.However,studies indicate that the pathogenesis of NAFLD may not be limited to high-fat,high-calorie diet,but also involves the interaction between environmental factors and host inheritance.CS may serve as one of the important factors.It is worth mentioning that the relationship between chronic psychological stress and NAFLD at the theoretical level of traditional Chinese medicine provides the possibility to reveal the effect mechanism of Sini Powder in the treatment of chronic psychological stress-related NAFLD,including the theory of emotional pathogenesis of traditional Chinese medicine,the theoretical viewpoint of"viscera and body function related",and the excellent effect of Sini Powder on chronic psychological stress disease and NAFLD in clinical.However,the mechanism of chronic psychological stress affecting NAFLD network is complex,and Sini Powder,as a traditional Chinese medicine compound,has a multi-target and multi-channel effect mechanism.Therefore,this study uses the methods of "network medicine" and "network pharmacology",combined with animal experiments,to predict,analyze and verify the core mechanism of chronic psychological stress inducing and aggravating NAFLD disease process from the system level,and the core mechanism of Sini Powder in the treatment of chronic psychological stress-related NAFLD.A total of 333 NAFLD related targets and 930 chronic psychological stress related targets were collected from the DiseaseGene Network database.After mapping NAFLD related targets to PPI network of chronic psychological stress,79 related interactive targets between chronic psychological stress and NAFLD were obtained,accounting for 23.7%of NAFLD targets.A PPI network with 79 nodes and 239 edges(protein interaction with threshold?0.7)was obtained in STRING Network database.Based on the topological analysis of PPI network in the software of Cytoscape,the average degree of 79 targets is about 6.05,and 31 hypothetical targets with topological importance are obtained by selecting targets whose degree is higher than the average,among which the inflammatory factor related targets account for 22.6%of the core targets,and the highest degree targets are IL6,TNF,IL1?.After enrichment analysis in the David database,21 related pathways(P<0.05)and 6 core pathways(P<0.01,FDR<0.01)were predicted to be involved in the induction and aggravation of NAFLD by chronic psychological stress NLRP3 may play a key role in the process of chronic psychological stress inducing and aggravating NAFLD.Sixty male Wistar rats(12weeks,250±10g)were randomly divided into four groups(n=8)according to the open field experiment at week 0 of the experiment,which were blank control group,chronic stress group(CS group),High fat diet group(HFD group)and Complex model group.The results of General observations showed there were significant depressive behavior in CS group and Complex model group.On the metabolic level,the rats in CS group,HFD group and Complex model group all showed significant pathological manifestations of NAFLD.Compared with the blank control group,the liver index(P<0.01),liver TG content(P<0.01),oil red O staining area(P<0.01),AST(P<0.01),ALT(P<0.05),TC(P<0.01),LDL(P<0.01),FFA(P<0.01)were significantly higher,and HDL was significantly lower(P<0.01)in CS group.In addition,compared with HFD group,liver weight,liver index,liver TG content,oil red O staining area(P<0.01),AST,ALT,FFA significantly increased(P<0.01)in complex model group.It shows that chronic psychological stress can induce and aggravate the disease process of NAFLD.On the nerve level,the blood GC content of CS group and Complex model group was significantly higher than that of blank control group and HFD group(P<0.01),and the relative expression of GR mRNA in hippocampus was decreased,but there was no significant statistical difference.On the inflammatory level,compared with the blank control group,the relative expression of NF-?B and NLRP3 mRNA in the liver of CS group increased significantly(P<0.01),the contents of NF-?B,NLRP3,ASC and caspase-1 in the liver increased significantly(P<0.01),and the contents of IL-1 ?,IL-6 and TNF-? in the liver and serum increased significantly(P<0.01)in CS group.Compared with HFD group,the relative expression of NLRP3 in liver was significantly increased(P<0.05),the contents of NF-?B,NLRP3,ASC and caspase-1 in liver were significantly increased(P<0.01),and the contents of IL-1 ?,IL-6 and TNF-? in liver tissue and serum were significantly increased(P<0.01)in complex model group.In addition,the correlation analysis showed that NLRP3 was significantly correlated with serum GC(P<0.01),and NLRP3,IL-1 ?,IL-6,TNF-? were significantly correlated with TG(P<0.01).The results show that chronic psychological stress can start and mediate NAFLD disease process through 79 related targets(31 of which are core targets)and 21 related action pathways(6 of which are core action pathways)through the "neuro inflammatory metabolic" network with inflammatory response as the core.The activation of NLRP3 and the release of inflammatory factors play an important role.In addition,NLRP3 and inflammatory factors may be the key targets for the treatment of chronic psychological stress-related NAFLD.The research further uses the method of network pharmacology and experimental verification to clarify the core effect mechanism of Sini Powder as the multi-target and multi-channel effect of traditional Chinese medicine compound in the treatment of chronic psychological stress-related NAFLD.779 compounds of Sini Power were collected in tcmsp,symmap,ETCM and npass databases After ADME parameters and Ribinsky's five rules screening,125 compounds with different chemical structures and druggability were obtained,and 311 potential targets of these compounds were found.20 interaction targets between Sini Powder and chronic psychological stress-related NAFLD were collected by gene mapping,accounting for 25.3%of the targets related to chronic psychological stress-related NAFLD disease,54.8%of the core targets.In addition the target of inflammatory factors accounted for 30%.A PPI network with 20 nodes and 80 sides was established in STRING database(Protein interaction with threshold?0.7).Through topological analysis,8 hypothetical targets with topological importance were obtained for the treatment of chronic psychological stress-related NAFLD by Sini Powder.Of the 8 core targets,50%were inflammatory factors.Enrichment analysis was carried out in David database.The results showed that there were 23 pathways(P<0.05),including 2 core pathways(P<0.01,FDR<0.01).Combined with the results of previous studies and Experiments 1 and 2,the analysis of "target pathway" network predicted by network pharmacology suggests that Sini Power may improve the inflammatory response of chronic psychological stress-related NAFLD by inhibiting NLRP3.Sixty male Wistar rats(12weeks,250±10g)were randomly divided into four groups(n=8)according to the open field experiment at week 0 of the experiment,which were blank control group,composite model group,Sini Power treatment group and positive drug treatment group.The general results showed that the composite model group,Sini Power treatment group and positive drug treatment group all showed significant depressive behavior,and Sini Powder could significantly improve depressive behavior after one week of treatment.On the metabolic level,compared with the blank control group,AST,ALT,ALP,TG,TC,FFA,LDL were significantly higher(P<0.01),and HDL decreased significantly(P<0.01)in the complex model group.After 1 week of treatment with Sini Powder and simvastatin,the liver weight,liver index,liver TG content,oil red O staining area,AST,alt,ALP,TG,TC,FFA,LDL and HDL in SNS group and positive group were significantly improved compared with those in complex model group(P<0.01).On the nerve level,the serum GC content of the complex model group was significantly higher than that in blank control group(P<0.01);the serum GC content of the SNS group was significantly lower than that in complex model group(P<0.01),and there was no significant change in the positive group compared with complex model group.On the inflammatory level,compared with the blank control group,the mRNA expression of NF-?B(P<0.01),NLRP3(P<0.05),ASC(P<0.01),caspase-1(P<0.01)in the liver of the complex model group increased significantly,and the protein content of NF-?B,NLRP3,ASC,caspase-1,IL-1 ?,IL-6,TNF-? in the liver and the content of IL-1?,IL-6,TNF-? in serum increased significantly(P<0.01)in complex model group.Compared with complex model group,the content of NF-?B,NLRP3,ASC and caspase-1 protein in liver decreased significantly(P<0.01),and the content of IL-1 ?,IL-6 and TNF-? in liver and serum decreased significantly(P<0.01)in SNS group and positive group.The results show that the effect of Sini Powder corresponds to the pathogenesis of chronic psychological stress-related NAFLD.125 compounds with different chemical structures and druggability in Sini Powder can improve the "neural-inflammatory-metabolic"network of chronic psychological stress-related NAFLD by acting on 20 related targets(8 core targets)and 23 related action pathways(2 core action pathways).The inhibition of inflammatory response mediated by NLRP3 and inflammatory factor may be the core.In addition,it also involves the regulation of insulin resistance,oxidative stress and other biological processes closely related to NAFLD. |
PDF Full Text Request