The Research On Hypoglycemic And Antibacterial Biological Activity Of Allium Fistulosum L

Allium fistulosum L was one of the common foods and complementary food flavors,and it was favored by people for its antibacterial,detoxifying and detoxifying effects.Chinese medicine believed that Allium fistulosum L was warm,spicy,with the effect of dispelling cold and stomach,phlegm,bactericidal,lungs,yang,sweating,bleeding,and pain relief.Scallions were widely used as medicine and food.At present,the biological activity research and practical application fields of Allium fistulosum L were relatively narrow.In order to more fully utilize and develop the natural resources of green onion,this study used Allium fistulosum L as raw material to optimize the ethanol extract(E001)of the onion by organic solvent extraction.The alcohol extract of Scallion was roughly divided into four solvent polar parts of petroleum ether extract(F001),ethyl acetate extract(F002),n-butanol extract(F003)and water soluble(F004),and ethanol extract of green onion.The in vitro hypoglycemic activity and antibacterial activity of each polar site were screened,and the mechanism of the action of F002 on the glucose metabolism of HepG2 cells in insulin resistance model was also discussed.The antibacterial activity of ethanol extract and polar parts of Scallion was studied.The antibacterial activity and chemical composition of the volatile components extracted from the green onion were optimized.(1)Taking the dried common onion as the raw material identified as Allium fistulosum L as the raw material,the total flavonoid content and the extract yield were evaluated.The ethanol extract(E001)was extracted by reflux extraction with ethanol assolvent.The three factors of ethanol concentration,solid-liquid ratio and reflux time were investigated by single factor experiment and orthogonal experiment.The optimal reflux extraction conditions were as follows: the ratio of material to liquid ratio was 1:20 with60% ethanol,The mixture was heated under reflux at 80 ? for 4 hours.The ethanol extract(E001)was obtained by the optimized extraction scheme,and the fractions were gradually extracted with petroleum ether,ethyl acetate and n-butanol to obtain four different polar sites,namely petroleum ether extract(F001)and ethyl acetate extract.(F002),n-butanol extract(F003)and water soluble fraction(F004).(2)Inducing insulin resistance cell model in HepG2 cells induced by high concentration glucose medium as an inducing factor,screening ethanol extract(E001)and four different polar sites by glucose oxidase method,and finally selecting to increase insulin resistance Ethanol extract(F002)of glucose uptake in cell model was used as an experimental object,and glucose of F002 against insulin resistance cell model was detected by glucose oxidase method,tetrahydronium salt colorimetric assay(MTT)and Western blotting(Western blot).Effects of consumption,cell viability,and protein expression of the insulin signaling pathway.RESULTS: Compared with the blank control group,when the glucose concentration in the induction medium was 40 mmol/L,the difference in absorbance was the largest,so 40 mmol/L glucose medium was selected to establish the insulin resistance cell model;F002 was used to treat insulin resistance cells.When the concentration is 80-20 mg/mL,the glucose consumption is slightly lower than that of the model control group;when the F002 concentration is 80-20 mg/mL,the cell survival rate can be kept above 80%,and the cytotoxicity is small;F002 can beRegulating the decrease in IRS-1 and IR? levels due to high glucose induction,reducing the expression level of p-IRS-1,and also preventing the decrease in GLUT2 levels.(3)8 kinds of broad-spectrum bacteria including Staphylococcus aureus,Klebsiella pneumoniae,Pseudomonas aeruginosa,Enterococcus faecalis,Acinetobacter baumannii,Bacillus subtilis,Escherichia coli and Bacillus thuringiensis,using filter paper method The ethanol extract and the high and low concentrations(0.1 g/mL and 0.05 g/mL)of different polar sites were tested for antibacterial activity,and the sites with antibacterial activity were further determined for minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC).Results: High and low concentrations of E001 and F001 had strong antibacterial activity against 8 kinds of bacteria,especially F001 was strong against Pseudomonas aeruginosa and Bacillus subtilis,MIC value was3.125 mg/mL,MBC value was 50 mg/mL.;F002 has obvious antibacterial effect on 8kinds of bacteria,especially against Klebsiella pneumoniae,Pseudomonas aeruginosa and Escherichia coli;F003 has the worst antibacterial effect,only pneumonia at low concentration Klebsiella has an inhibitory effect,and at high concentrations,it has no inhibitory effect on Pseudomonas aeruginosa and Enterococcus faecalis.F004 inhibits bacterial growth,but it has the most obvious inhibitory effect on Staphylococcus aureus.(4)Ultrasonic-assisted steam distillation was used to extract the volatile components of the onion.The extraction rate of volatile components was evaluated.The ultrasonic time,solid-liquid ratio and extraction time were investigated by single factor experiment and orthogonal experiment.GC-MS identified its main chemical components and evaluated the antibacterial activity of the volatile components of the onion.RESULTS:The best extraction process for volatile components of green onion was ultrasonic 40 min,ratio of material to liquid 1:3,distillation time 3 h.The actual extraction rate of volatile components of green onion was 0.0416%.Through GC-MS analysis,NIST mass spectrometry library search identified 42 major compounds,accounting for 99.99% of the total volatile components.The peak area normalization method was used to determine the content of each component.The main substance was sulfur compounds.,the content of more is fluorocytosine,allyl isopropyl sulfide,propylene methyl sulfide,dimethyl trisulfide,dipropyl trisulfide,tetrasulfide,thiophene,thiazane,thiazole class.The biological activity evaluation experiment showed that the volatile components of the onion had good activity against the selected 8 strains.(5)Using starch and dextrin as auxiliary materials,90% ethanol as a wetting agent to granulate the total extract of green onion ethanol,and study the preparation process of hard onion extract hard capsule.RESULTS: According to the best extraction process,a large amount of extraction was carried out.After removing impurities with purified water,the ethanol was recovered and dried in a vacuum drying oven to constant weight.16.2 g of dried onion total extract powder was weighed,4.9 g of starch was added,and 4.9 g of dextrin was added.,mixing,wet granulation with 90% ethanol as wetting agent,vacuum drying at 60 °C,granulation,passing through 40 mesh sieve,200 mesh sieve to remove fine powder,and loading the powder particles under the environment of humidity below60% Capsule No.3,granulating 100 capsules,measuring 5 mg of total flavonoids per capsule.Serve the onion extract capsule.