Study On The Flavonoids Content Of Hawthorn Leaves And The Activity Of Regulating Glycolipid Metabolism

Disorder of glycolipid metabolism is an important risk factor for cardiovascular and cerebrovascular diseases.Clinical treatment needs to use a variety of drugs at the same time,which the side effects of long-term drug use are obvious.It is of positive significance to find safe and effective drugs for the prevention and treatment of glycolipid metabolic syndrome in order to reduce the risk of cardiovascular and cerebrovascular diseases,diabetes and its complications.Hawthorn leaves have the effect of regulating glucose and lipid metabolism The planting area is scattered where the climatic conditions are obviously different.In this paper,a HPLC method for the simultaneous determination of six flavonoids was established to investigate the quality of Hawthorn leaves from different producing areas,different varieties and different years.the activity of vitexin glucosides(VOG)isolated from Hawthorn leaves and hypericin(HP),a quality control component of Hawthorn leaves regulated by Chinese Pharmacopoeia,were studied using insulin resistance(IR)HepG2 cell model.The protein and gene expression of adenosine monophosphate activated protein kinase(AMPK),acetyl coenzyme A carboxylase(ACC)and glucose transporter 4(GULT4)were measured by western blotting and Real Time-PCR.This provides basic data for comprehensive evaluation of Hawthorn leaf medicinal materials resources,development and application1.The contents of six flavonoids including VOG,vitexin rhamnoside(VOR),vitexin rhamnoside(VT),rutin(RU),HP,luteoloside and total flavonoids(HLF)in Hawthorn leaf collected in October from 2014 year to 2018 year,from Weifang city,Shandong Province,Yuncheng city,Shanxi Province and Chengde city,Hebei Province,were investigated.The results showed that the composition,proportion and content of flavonoids in hawthorn leaves were significantly affected by the origin and harvest time.The content of VOG,VOR,VT accounted for 60%of the total content of 6 flavonoids.The HLF content of Crataegus pinnatifida Bge.var.major N.E.Br.collected from Chengde city Hebei province was the highest,followed by Crataegus cuneata Sieb.et Zucc.2.Three components were isolated from Hawthorn leaves and were identified to be VOG,VOR,VT.VOG and HP had no effect on the proliferation and activity of HepG2 cells at the concentration of 0.125 ? M-32 ? M.Compared with the model group,VOG and HP could promote glucose consumption and decrease the levels of the lipid droplet accumulation,TG,total cholesterol,leptin and adiponectin in IR HepG2 cells.3.The protein and gene expression of AMPK,ACC and GULT4 in IR HepG2 cells treated with VOG and HP were detected by western blotting assay and Real Time-PCR assay.The results showed that both VOG and HP could increase the protein ratio of p-AMPK/AMPK and p-ACC/ACC,and increase the GULT4 protein level.Both HP and VOG could down-regulate the mRNA expression of AMPK and ACC,and up-regulate GULT4 mRNA expression in a concentration-dependent manner.This suggested that VOG and HP might regulate glycolipid metabolism by activating AMPK signal pathway.