| The uygur medicine gulina,also known as pomegranate flower,is the dried petals of Punica granatum L.,a deciduous shrub or small tree of the family Pomegranate.The chemical composition of pomegranate flowers category contains polyphenols,flavonoids,triterpene,saponin and so on.The main active ingredients of pomegranate flower are gallic acid,ellagic acid,corilagin,anthocyanin,oleanolic acid,tricin,daucosterol,etc,and they have convergence and hidroschesis hemostatic effect.In the modern research of pomegranate flowers,In the modern research of pomegranate flower,it is found that it has good anti-type 2 diabetes and complications,protection of circulatory system,anti-inflammatory,anti-oxidation,liver protection and so on.In the early stage,our research group separated and identified the compounds in the extraction sites of ethyl acetate,n-butanol and petroleum ether of pomegranate flowers.However,there are few studies on the active ingredients and mechanism of action of pomegranate flowers.Therefore,this article uses type 2 diabetes and hyperlipidemia as models to study the active ingredients and mechanism of the pomegranate flower’s hypoglycemic and lipid-lowering effects.Objective :(1)To compare the effects of different extraction parts of pomegranate flowers on insulin resistance and study the mechanism of action.(2)To study the active components and mechanism of lipid lowering in pomegranate flower extract.Methods :(1)A high-fat diet combined with streptozotocin was used to establish a rat model of type 2 diabetes.Metformin and pomegranate flower extracts were given for 4 weeks,and serum insulin content and other biochemical indexes were measured.(2)The effects of extracts from different parts of pomegranate flowers on glucose intake and related protein expression in insulin-resistant 3T3-L1 adipocyte were determined.(3)Network pharmacology was used to study the active ingredients andmechanism of reducing blood lipid in pomegranate flowers.(4)Mice hyperlipidemia model was established by gavage with high-fat emulsion,pomegranate flower extract and its active components were given for 8 weeks,and serum and liver lipids as well as the expression of PPARα in liver and PPARγ root were determined.(5)On the basis of the 3T3-L1 preadipocyte,effects of extracts from pomegranate flowers and their compounds on cell proliferation were determined by cck-8 method.The effects of extracts from pomegranate flowers and their compounds on cell differentiation were studied by drug administration in the process of cell differentiation,and intracellular triglyceride content and lipid gene expression were determined after differentiation.Hoechst staining and flow cytometry in combination with Annexin V/FITC PI staining to study the effect of pomegranate flower extract on cell apoptosis.Results:(1)The pomegranate flowers different extraction parts can reduce the type 2 diabetic rats fasting blood glucose,serum insulin,blood lipid level,oxidative damage degree and the insulin resistance index,improve the insulin sensitive index,and improve the insulin resistance of 3T3-L1 cells glucose intake,improve the p-Akt,PI3 K,Glut4 protein expression and water part extract improve the PPARγprotein expression.(2)Five active ingredients and 105 target genes of pomegranate flower for lowering blood lipid were obtained through network pharmacology.The active ingredients were C1,C4,C2,C3 and keampferol.Biological processes were response to lipid regulation and positive regulation of metabolic process.The KEGG pathways were mainly PI3K-Akt,AMPK,MAPK,apoptosia and PPAR signaling pathways.(3)It was found in animal experiments that the extract and five compounds had the effect of lowering blood lipid,among which C2 had the best effect.(4)Pomegranate flower extracts promote or not affect 3T3-L1 preadipocyte proliferation and differentiation in low dose,and can reduce apoptosis 3T3-L1 preadipocyte induced by palmitate,a reduction in the number of early and late apoptosis cells,improve the Bcl-2 m RNA and protein expression of Bax,Caspase 3 m RNA and protein expression.Medium and high dose pomegranate flowers,C1,C2,C4,keampferol treatment to different extent inhibited 3T3-L1 preadipocyte proliferation and differentiation,reduce the accumulation of lipids in the cell,reduce fat factor expression,and the expression ofPI3 K protein,p-Akt inhibition of PI3K-activation of Akt pathway were inhibited,PPARγ protein expression was reduced.Conclusion: Pomegranate flower extract can improve insulin resistance by activating PI3K-Akt pathway,and water extract can also improve insulin sensitivity by activating PPARγ receptor.C1,C2,C3,C4 and keampferol in pomegranate flowers all had a certain effect on lowering blood lipid,among which C2 had the best effect on lowering blood lipid in animal experiments.Pomegranate flower has a concentration-related bidirectional effect on the proliferation,differentiation and apoptosis of 3T3-L1 preadipocytes,which mainly depends on the regulation of PI3K-Akt pathway.This study is the first to study the regulation effect of pomegranate flowers on the PI3K-Akt pathway,which has guiding significance for the treatment of type 2 diabetes,hyperlipidemia,obesity and other diseases of pomegranate flowers. |
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