Basic Research On Hypoglycemic And Antibacterial Substances In Zingiber Striolatum Diels

Zingiber striolatum Diels,a perennial herb of the genus Zingiberaceae,is a kind of perennial herb Zingiber striolatum Diels.It can be directly cooked and eaten,and is rich in dietary fiber,protein and essential amino acids.More and more scholars have begun to study Zingiber striolatum Diels.Zingiber striolatum Diels has the functions of lowering blood sugar,inhibiting bacteria and treating dysmenorrhea,but the related substance base and its role in pharmacodynamics is still unclear.At present,the application field of Zingiber striolatum Diels is relatively narrow,especially in the research and application of its biological activity.In order to make better use of Zingiber striolatum Diels' s natural resources,this study used Zingiber striolatum Diels as raw material to ethanol extract and volatile components,and analyzed the chemical constituents of volatile components from Zingiber striolatum Diels,and carried out the ethyl acetate extract of Zingiber striolatum Diels.Simple isolation and purification were carried out to screen the anti-diabetic activity of ethanol extract and polar parts in vitro,and to explore the mechanism of ethanol extract and its polar sites on glucose metabolism in HepG2 cells.Finally,the antibacterial activity of ethanol extract and its polar parts in vitro was studied.(1)Orthogonal experiment was used to study the effects of ethanol concentration,the ratio of material to solution and extraction time on the content of total flavonoids and the extraction rate of total extract from Zingiber striolatum Diels.The results showed that the content of total flavonoids from Zingiber striolatum Diels,the best was 70% ethanol,the ratio of material to solution of 1:12,and the extraction time was 2h.(2)Single factor experiment,multi factor and multi level orthogonal experiment and ultrasonic assisted distillation were used to extract volatile components from Z.striolatum Diels.Taking the extraction rate of the Z.striolatum Diels volatile components as the evaluation index,the effects of ultrasonic time,distillation time and solid-liquid ratio on the extraction rate of volatile components were studied respectively,The chemical constituents and types of volatile components are analyzed by gas chromatography-mass spectormetry(GC-MS)and NIST4.0 mass spectrometry.The relative percentile content of each component were calculated by using peak area normalization method.The optimal extraction process of the volatile components were sultrasonic 60 min,distillation time 4h,material liquid ratio 1:3g/ml,extraction rate reached 0.0368%;and Identified 17 major chemical composition,which accounted for 85.074% of the total volatile oil,of which were eight kinds of alkenes,and three of the highest content were palmitic acid(30.508%),beta parsley(12.359%)and erucate(7.376%).(3)The Zingiber striolatum Diels was washed and dried,and the powder is filtered through a 60 mesh sieve to obtain a Zingiber striolatum Diels powder.70% ethanol,ratio of material to liquid 1:12,reflux extraction for 2h,to obtain ethanol extract(E001),and then extract E001 with petroleum ether,ethyl acetate and n-butanol to obtain petroleum ether extract(F001),acetic acid B Ester extract(F002),n-butanol extract(F003)and water soluble(F004).The F002 was isolated and purified by silica gel column chromatography with thin layer chromatography,isolation and identification of thymidine in the first time from Zingiber striolatum Diels.(4)Inducing insulin resistance cell model in HepG2 cells induced by high concentration glucose medium as an inducing factor,and detecting the glucose consumption of the Zingiber striolatum Diels ethanol extract and various polar partsof the insulin resistance cell model by glucose oxidase method,tetrazolium salt Cell viability was measured by colorimetric assay(MTT).The experimental results showed that the ethanol extract of Zingiber striolatum Diels had hypoglycemic activity,and the effect of protein expression of insulin signaling pathway was analyzed by Western blot: 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;Glucose consumption was significantly higher than other polar sites and blank control group,but lower than the positive control group;cell survival rate could be maintained at about 85%,and cytotoxicity was small;cation ethanol extract against IRS-1,IR? The expression levels of P-IRS-1 and GLUT2 proteins have a certain enhancement effect.(5)The antibacterial activity of different polar extract was measured with a filter paper,and the minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)were further measured at the sites with antibacterial activity.The experimental results showed that the Zingiber striolatum Diels E001?F001?F002?F003?F004had significant effects on Staphylococcus aureus,MIC value between 3.125-25mg/mL,MBC value 25mg/mL,Among them,petroleum ether extract and ethyl acetate had the best antibacterial activity against Staphylococcus aureus;the Zingiber striolatum Diels ethanol extract,petroleum ether extract,ethyl acetate extractand n-butanol extract had strong antibacterial activity against Bacillus subtilis,MIC values between 3.125-25 mg/mL and MBC values of 25 mg/mL,Among them,ethyl acetate extract has the best antibacterial activity against Bacillus subtilis;ethanol extract and petroleum ether extract from the Zingiber striolatum Diels had antibacterial effects against Escherichia coli,Klebsiella pneumoniae,Enterococcusfaecalis,Pseudomonas aeruginosa,and Acinetobacter baumannii,Petroleum ether extract has better antibacterial activity;ethanol extract,petroleum ether extract,ethyl acetate extract,n-butanol extract and water extract hadn't bacteriostatic effect on Bacillus thuringiensis.In order to provide theoretical basis for the in vitro antibacterial activity and natural food preservatives of Zingiber striolatum Diels.