| Purpose:Network pharmacology was used to predict the potential molecular biological mechanism of huanglian jiedu decoction in preventing and treating lipid metabolism disorders,and to investigate the effect and mechanism of heat-clearing and detoxification on lipid metabolism and inflammation in liver of hyperlipidemia rats.Material and method:1.Through the traditional Chinese medicine system pharmacology analysis platform(TCMSP),all the chemical components and active components related to the 4 traditional Chinese medicine in huanglian jiedu decoction were found and the drug molecule-target network was constructed.TTD,Dis Ge NET and Drug Bank databases were used to screen hlp-related targets,and a protein-protein interaction network of drug-disease targets was constructed by using STRING database(functional protein association network database).The core targets were identified for network topology analysis,and the core target proteins were analyzed by GO analysis and KEGG pathway enrichment analysis using DAVID database.2.40 SPF grade healthy SD rats were fed water freely and were randomly divided into normal group,model group,coptis antidotal decoction group and simvastatin group on average after7 days of adaptive feeding,with 10 rats in each group.The normal group was given normal diet daily,while the model group,the huanglian jiedu decoction group and the simvastatin group were given high-fat diet daily,free feeding and drinking water.Eight weeks later,the model group,huanglian jiedu decoction group and simvastatin group to continue feeding high-fat feed,at the same time to fill the stomach,to medicine dose conversion coefficients according to the equivalent dose conversion,huanglian jiedu decoction group was given traditional Chinese medicine yinpian,270 mg/kg/day a lavage,simvastatin group simvastatin for a 1.8 mg/kg/day a lavage,model group,the daily volume of saline lavage.After 4 weeks of treatment,serum lipid levels of TC,TG,LDL-c and HDL-c were detected.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of the liver tissues in each group,and the lipid deposition in the liver tissues in each group was observed by oil red O staining(ORO)method.The expressions of NLRP3,caspase-1,IL-1β,IL-18,and protein in liver were detected by Western Blot and the m RNA expression of NLRP3,caspase-1,GSDMD,IL-1β,IL-18,TNF-α,and genes in liver were detected by RT-PCR.Results:1.According to the screening conditions OB(oral bioavailability)≥ 30% and DL(drug similarity)≥0.18,34 Cortex phellodendri,16 Gardenia Jasminoides,14 Rhizoma coptidis and 36 Radix Scutellariae were screened for the active ingredients of the 4 drugs in huanglian jiedu decoction.There are 254 hyperlipidemia disease targets in TTD database,Dis Ge NET database and Drug Bank database.There were 41 mapping targets of huanglian jiefu decoction group and disease targets,mainly involving CRP,TNF,IL1 B,IL6,MAPK1,NOS3,VCAM1,etc.According to GO function enrichment analysis,GO items are divided into three categories,among which 809 items are biological process(BP),98 items are Cellular Components(CC)and 231 items are molecular function(MF).KEGG enrichment results showed that the top 30hlp-related pathways were selected according to PValue ascending sequence,including nod-like receptor signaling pathway,TNF signaling pathway,cell Apoptosis pathway,Fox O signaling pathway,and Platelet activation pathway.Nod-like receptor signaling pathway is the main inflammatory response pathway in the NLRs family,and the pyroptosis involved by NLRP3 inflammomes is also in this pathway.2.The results of blood lipid level showed that compared with the normal group,TC,TG and LDL in the model group were significantly higher than those in the normal group(P < 0.01),and HDL was significantly lower than those in the normal group(P < 0.01).Compared with the model group,the levels of TC,TG and LDL in the huanglian jiedu decoction group and simvastatin group were significantly reduced(P < 0.01)and the levels of HDL were significantly increased(P < 0.05).3.HE staining showed that: compared with the normal group,the model group had a large number of fat vacuoles in the liver.Compared with the model group,the liver fat vacuoles in the huanglian jiedu decoction group and the simvastatin group were significantly reduced.4.The results of oil red O staining showed that: compared with the normal group,the liver cell structure of the model group was not clear,and a large number of red lipid droplets of different sizes were seen in the visual field,with lipid droplets aggregation.Compared with the model group,the cytoplasm of huanglian jiedu tang group and simvastatin group was light blue,and the number of red lipid droplets was significantly reduced.5.Western blot detection of liver NLRP3,caspase-1,IL-1β,IL-18 protein expression results:compared with the normal group,the model group showed significantly higher liver NLRP3,caspase-1,IL-1β,IL-18 protein expression levels than the normal group(P < 0.01).Compared with the model group,the expression levels of NLRP3,caspase-1,IL-1β and IL-18 proteins were significantly decreased in the treatment group and simvastatin group(P < 0.01).6.Rt-pcr was used to detect the expression levels of NLRP3,GSDMD,caspase-1,TNF-α,IL-1β,and IL-18 m RNA in liver(P<0.01).Compared with the model group,the expression levels of NLRP3,GSDMD,caspase-1,TNF-α,IL-1β,and IL-18 m RNA in the treatment group and simvastatin group were significantly reduced(P < 0.01).Conclusion:1.Huanglian jiedu decoction plays an anti-hlp role through multi-component,multi-target,synergistic regulation of nod-like receptor signaling pathway,lipid metabolism,inflammatory response,oxidative stress and other signaling pathways.2.Huanglian jiedu decoction can effectively regulate the lipid level of hyperlipidemia rats,reduce inflammatory factors,and improve liver tissues and cells.3.Huanglian jiedu decoction improves the mechanism of liver tissue damage in hyperlipidemia rats,which may be related to the inhibition of nlrp3-mediated inflammatory response and cell pyroptosis. |
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