Sdg Extraction And Protect The Kidney Of Diabetic Mice

Plant lignans are phenolic secondary metabolites whose basicstructure is a 2,3-dibenzylbutane. They possess oestrogenic/anti-oestrogenic effects. Secoisolariciresinol diglucoside (SDG) is animportant dietary lignan that is found at very high levels in flaxseed (1—4%, w/w). Flaxseed lignans have received much research interest in recentyears because of reported phyto-oestrogenic, anticarcinogenic, andanti-atherogenic effects. Studies of both type 1 and 2 diabetes modelshave reported positive results when using SDG. There are many possiblemechanistic explanations for the observed bioactivities includinginvolvement in hormonal metabolism or availability, angiogenesis,anti-oxidation and gene suppression. But the biology function andprotective effects on Diabetic Mellitus have not been reported in China.Therefore, we establish a sensitive analytical method for detectionsecoisolariciresinol(SECO) in flaxseed using high performance liquidchromatography(HPLC) and extract lignans from flaxseed. Explore theeffects of flaxseed lignan extracts and its mechanism on early renallesions of diabetic rats.PartⅠDetermination of secoisolariciresinol in flaxseed by HPLC The aim was to establish a sensitive analytical method for detection ofsecoisolariciresinol(SECO) in flaxseed using high performance liquidchromatography(HPLC). Orthogonal test was used to evaluate theinfluence of 4 factors, including the extracting concentration ofmethanol and the time, temperature, concentration of HCl hydrolysis.HPLC method was used to determine the SECO in flaxseed. Theoptimum extraction was as follows: 90% methanoland, the temperature,time, concentration of HCl hydrolysis is 90℃, 1h, 3.5mol/L. The linearitywas superior from 14.8-74μg/ml. In flaxseed the content of SECO wasabout 1.56mg/g, the standard deviation was 3.47%. The average recoveryrate was 92.7% and 98.9%. The results suggest that this method is quickand economical. Also it has been satisfactory applied to the routineanalyses.PartⅡStudy on Extracting Technologies ofLignans from FlaxseedTo investigate the technological parameters of the extraction processof lignans from flaxseed. The 70% ethanol or 90% methanol withultrasonic wave or water-bath technology were used. The parameters ofextractive weight and the contents of lignan were compared respectively.Using C18 column, the active component is isolated and purified.Identification of the component was complished by UPLC/MS. The results showed the alcohol extract method with ultrasonic wavetechnology was best. The content of lignan was 201mg/g. C18 columnwas considered as a suitable medium for purification of SDG.PartⅢStudy on the Protective Effect of Flaxseed LignanExtract on Kidney in Diabetic RatsThe purpose of this study is to investigate the protective effect ofdiaetic mice. ICR mice were randomly divided into 6 groups: normalmice control, STZ diabetic mice control, STZ diabetic mice fed with FLEat dose of (600mg/(kg·d), 1.2g/(kg·d), 3.6g/(kg·d)), and STZ diabeticmice fed with metformin. The above components were administedintragastrically for 8 weeks. Then their blood glucose, kidney weight tobody weight, 24-hour urinary protein were detected. Glomerular structureof kidney were examined by light microscopy. The results were that thekidney weight to body weight, 24-hour urinary protein in high,middledose of FLE groups were significantly lower than those in diabeticcontrol group(P＜0.05). The content of MDA in all dose of FLE groupswere significantly lower than those in diabetic control group(P＜0.05).And the activities of SOD, GSH-Px in high, middle dose of FLE groupswere significantly enhanced than those in diabetic controlgroup(P＜0.05). The results indicate that FLE has no effect in decreasingblood glucose, while it can ameliorate the renal function maybe by inhibiting the oxidative stress of kidney.PartⅣA Experimental Study on Acute Toxicity ofthe Flaxseed Lignan ExtractTo study acute toxicity of the flaxseed Lignan Extract on mice andoffering evidence for clinical safe dose. An acute toxicity test wasconducted. LD50 was carried out by stomach feeding with 7 dosagegroups, and the mice were observed for 14 days after first amount wasgiven to the mice by stomach feeding. The behavioral indexes andtoxicity degree were continuously observed. Then in the end of the test,the mice were executed in order to get primary toxicity materials. Allmice healthily survived during the period of test, and no obvioussymptoms were observed. The LD₅₀ dose is more than 21.500g/kg. Thedrug had no harmful effect on body weight as well as organ coefficients.Observed by naked eyes, there were no abnormal changes on surfaces andtransects of organs. The results of pathological examination on all organswere also normal. The results indicate that no acute toxicity is found with,and it may be safe to be used in clinic.