Effects Of Red Raspberry And Its Extracts On Fat Accumulation In HepG2 Cells

Red raspberry is mainly cultivated in the northeastern part of China.It is rich in a variety of biological active ingredients.At the same time,it has high medicinal,edible and therapeutic value,and has a certain positive effect on the regulation of lipid metabolism.In this paper,the content of active ingredients and the composition of polyphenols in red raspberry juice,red raspberry wine,red raspberry extract,seed extract and the effects of red raspberry juice,red raspberry wine,red raspberry extract,seed extract,ellagic acid and raspberry ketone on the fat accumulation of HepG2 cells were studied.The main conclusions are as follows:The contents of total phenolic,flavonoids,proanthocyanidins and anthocyanins in red raspberry extract,red raspberry seed extract,red raspberry juice and red raspberry wine were determined.The results showed that the contents of total polyphenols,flavonoid,proanthocyanidins and anthocyanins in red raspberry juice were 14.90±0.78 mg GAE/g,7.45±0.52 mg RE/g,75.42±3.79 mg CE/g,3.53±0.26 mg CGE/g;in red raspberry wine were 35.01±2.06 mg GAE/g,21.21±1.03 mg RE/g,149.20±4.90 mg CE/g,4.69±0.32 mg CGE/g;in red raspberry extract were 15.18±0.94 mg GAE/g,1.25±0.05 mg RE/g,20.35±1.06 mg CE/g,1.15±0.70 mg CGE/g,respectively;in raspberry seed extract were 15.35±1.55 mg GAE/g,11.22±0.66 mg RE/g,27.08±0.99 mg CE/g,but anthocyanins not detected.Ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was used to analyze the active ingredients in red raspberry juice,red raspberry wine,red raspberry extract and seed extract.The results showed that six active ingredients such as rutin,quercetin,isorhamnetin,gallic acid,caffeic acid and ellagic acid were identified in red raspberry juice,red raspberry wine,red raspberry extract and seed extract.The total content of polyphenols in raspberry wine was the highest.Red raspberry juice contained 10 active ingredients such as rutin,catechin,cryptochlorogenic acid and isorhamnetin.Red raspberry wine contained 9 kinds of active ingredients such as gallic acid,quercetin,ellagic acid and caffeic acid.Red raspberry extract contained 13 active ingredients such as catechin,epicatechin,rutin and ellagic acid.Red raspberry extract contained 12 active ingredients such as epicatechin,catechin,ellagic acid and quercetin.Oil red O staining method was used to compare the fat accumulation of HepG2 cells which were induced by different oleic acid modeling agents with the lipid content and triglyceride(TG)content as indicators,in order to screen out suitable modeling agents to establish a fat accumulation model.The results showed that oleic acid modeling agent A with dimethyl sulfoxide(DMSO)as solvent was selected to have a concentration of 500 ?mol/L,at this time,the intracellular lipid content and TG content were 1.75 times and 1.71 times of that in the control group;the oleic acid modeling agent B with bovine serum albumin(BSA)as the solvent was 1000 ?mol/L,at this time,the intracellular lipid content and TG content were 2.10 times and 1.93 times of that in the control group respectively;no significant fat accumulation of the cells was observed after the alcohol-forming agent induced the cells;finally,1000?mol/L oleic acid modeling agent B was used to induce HepG2 cells to establish a fat accumulation model.The results of the MTT assay showed that red raspberry extract at a concentration of 200?g/mL,red raspberry seed extract at 4 mg/mL,2%red raspberry juice,1%red raspberry wine,40 ?mol/L ellagic acid and 250 ?mol/L raspberry ketone were used as the upper limit mass concentration of red raspberry and its active ingredients in the fat accumulation of HepG2 cells.Red raspberry extract could significantly reduce hepatocyte fat accumulation induced by 1000 ?mol/L oleic acid.When 200 ?g/mL red raspberry extract induced cells,the intracellular lipid content decreased by 25.93%,TG content decreased by 37.23%,LDL-C content decreased by 32.03%and HDL-C content increased by 32.36%in comparison with the model group.When 4 mg/mL raspberry seed extract induced cells,the intracellular lipid content decreased by 47.12%,TG content decreased by 46.23%,LDL-C content decreased by 35.32%,HDL-C content increased by 35.51%in comparison with the model group.When 2%red raspberry concentrated juice induced cells,the intracellular lipid content decreased by 27.61%,TG content decreased by 27.20%,LDL-C content decreased by 23.30%and HDL-C content increased by 23.60%.When 1%red berry wine was treated,the intracellular lipid content decreased by 15.13%,TG content decreased by 11.22%,LDL-C content decreased by 11.33%and HDL-C content increased by 12.36%.When treated with 40 ?mol/L ellagic acid,the intracellular lipid content decreased by 29.64%,TG content decreased by 29.40%,LDL-C content decreased by 20.09%and HDL-C content increased by 14.61%.When treated with 250?mol/L raspberry ketone,the intracellular lipid content decreased by 23.18%,TG content decreased by 28.24%,LDL-C contained decreased 13.64%and HDL-C content increased by 19.27%.It indicated that red raspberry and its extract can reduce the accumulation of fat in HepG2 cells.