Sutdy On Extraction And Anti-inflammatory Effect Of Anthocyanin Extracted From Raspberry

Raspberry contains a variety of functional ingredients,such as anthocyanins,ellagic acid,flavonoids and other phenols,salicylic acid,vitamin C,etc.It has a high research value and food value.Anthocyanins are natural pigments that we all know for their ability to resist oxidation and scavenge free radicals.Some studies have shown that anthocyanins are toxic to fungi,yeasts,bacteria and other microorganisms.They can prevent DNA mutations,and have anti-cancer,anti-inflammatory and other functions.In this paper,the content of anthocyanin in raspberry was determined by HPLC.The HPLC system was:mobile phase A:acetonitrile:methanol:formic acid:water=22.5:22.5:8.5:41.5(V/V/V/V);mobile phase B:formic acid:water=8.5:91.5(v/v).Gradient elution,0 min,7%A;25 min,40%A;50 min,93%A.Detection wavelength:535 nm;flow rate:1.0 mL/min;column temperature 30?;each injection volume was 10 ?IL.The results showed that the anthocyanin was not contained in the raspberry leaves from the seedling stage to the fruiting stage.For the berry fruit,the single factor experiment was used to measure the yield of the anthocyanin.It was 119.2269 ?g/g,when the extraction time was 70 min.when the ratio of material to liquid was 1:10,the highest yield of anthocyanin was 133.8927 ?g/g.When the concentration of methanol was 80%,the highest yield of anthocyanin was 119.9516 ?g/g.Further orthogonal experiments were conducted to obtain the best process for extracting anthocyanins.It was that the extraction time was 70 minutes,the methanol concentration was 90%,and the ratio of solid to liquid was 1:11.The final yield of anthocyanins was 156.2897?g/g.In this study,mouse myeloid macrophages(BMDM)were used as materials to study the effects of anthocyanins on the expression and production of inflammatory cytokines related to BMDM.The effect of different concentrations of anthocyanins on the activity of BMDM cells was tested by MTT assay.The results showed that treatment of BMDM cells with 1-30 ?g,mL anthocyanin for 24 h had no effect on the proliferation of BMDM in vitro.Effects of anthocyanins on LPS(1.0 ?g/mL)stimulation of BMDM-transformed M1 macrophages was texted.After treatment of M1 macrophages with 5 ?g/mL,10?g/mL and 20 ?g/mL anthocyanins respectively,the test results showed that anthocyanin can down-regulate the expression and production of TNF-?,IL-6,IL-1?,MCP-1,iNOS and other M1 macrophage marker molecules,and different concentrations of anthocyanin on different Cytokines have different effects and the results of mRNA and protein levels are consistent.The effect of anthocyanins on the conversion of M2 macrophages to BMDM cells stimulated by IL-4(20 ng/mL)was treated with 5 ?g/mL,10 ?g/mL,and 20?g/mL anthocyanins respectively.The phagocytes showed that the anthocyanins up-regulated the expression of IL-10 and Arg-1 marker molecules of M2 macrophages and the secretion of production factors.Anthocyanins of different concentrations had different effects on different cytokines.The effect was basically a dose effect,and the results of mRNA and protein levels were consistent.The above results suggest that anthocyanins can inhibit the secretion of the M1 macrophage marker and promote the secretion of the M2 macrophage marker,indicating that the anthocyanins have significant anti-inflammatory effects and are associated with inflammation.Metabolic diseases have preventive and palliative effects.