| Objective: Using three different methods of semi-bionic, waterextraction and ethanol extraction and comparing those extractionmethods, extraction technology of corn-silk total saponins wasresearched, so as to provide a basis of industrialization andtechnology for corn-silk total saponins. Adopting purifiedcorn-silk total saponins for treatment diabetic mice to explore theprevention of diabetes complications.Methods:1. Total Saponins Extract Process Comparison of Stigma MaydisUltraviolet visible spectrophotometry was used to determine thecontent of total saponin in Stigma Maydis. We studied the effectof single factor such as acid extraction time, base extraction time,ultrasonic power, liquid solid ratio to the extraction rate of totalsaponin. orthogonal experiment method was used to optimize theprocess parameter of Semi-bionic, water extraction and ethanolextraction, and extraction ratio of total saponin using thesemethods were compared.2. Purification Process of Total Saponins Extract in Stigma MaydisMacroporous adsorbing resin D101was used to purified totalsaponins by ethanol extraction. We discussed the purificationprocess in the aspects of extract concentration, flow velocity, ethanol eluent in different concentration, etc.3. Diabetic Complication Prevention of Stigma MaydisKunming mice (Certificate of Conformity: SCXK (hubei)2008-0003),male, weighing18-22g, provided by the Wuhan Institute ofBiological Products.50mice were treated with Streptozotocin200mg/kg by intraperitoneal injection after overnight fasting of12h. Fasting blood-glucose was detected3days later.Streptozotocin-induced diabetic mice model used the mice withblood-glucose value between14to20mmol/L. Model mice wererandomly divided into3groups. Positive control mice were treatedwith rosiglitazone1mg/kg by oral gavage. Experimental group micewere treated with total saponin extract of Stigma Maydis400mg/kgby oral gavage, and high fat diet for daily diet. Wild Type micewere treated with ordinary diet. After four weeks of oral gavage,collecting the Orbital blood to determine the related index.Results:1. The optimized extract process of Semi-bionic was A2B2C2D3,acid(pH=2.5）extraction time was90min, base（pH=7.5）extractiontime was90min, ultrasonic power was70W, solid liquid ratio was1:40，Effect of factors were in the order of A acid extraction time>Dsolid liquid ratio>C ultrasonic power> base extraction time. Theextraction rate of total saponin was0.676%.The optimized condition of water extract was A3C2B2, extractiontime was120min, ultrasonic power was70W, solid liquid ratio was1:30，the effect of factors were in the order of A>C> B, Theextraction rate of total saponin was0.329%. The optimized condition of ethanol extract was A1C3B3D3, ethanolconcentration was60%，extraction time was120min, ultrasonic powerwas90W, solid liquid ratio was1:40，the effect of factors werein the order of D>C>A>B, The extraction rate of total saponin was0.564%.The extraction rate of total saponin of Semi-bionic was highest.The reason are likely the acid-base interaction leading StigmaMaydis Cell wall rupture, which result in a large number of activecomponent dissolving.2. Optimized condition of Macroporous adsorbing resin D101was：sample volume was20ml, flow was2ml/min,sample concentration was44.96mg／ml, optimized eluent concentration was70%ethanol. Theextraction of total saponin in this condition was36.91mg/ml, andthis process was stable and efficient.3. The average value of blood glucose of model mice was16.2mmol/L.The distinct difference（P<0.01） between model and wild typeindicated modeling successfully.Blood glucose index：experimental group(total saponin of StigmaMaydis), positive group(rosiglitazone) were distinctivelydecrease the blood glucose index compared with WT group （P<0.01）,indicating that total saponin of Stigma Maydis has a effect ofdecreasing diabetic bloof glucose.TC and TG index：The distinct difference between model and WTindicated streptozotocin diabetes mice initiating lipid metabolicdisorder. The distinct difference between experiment group and WTindicated total saponins of Stigma Maydis can ameliorate lipidmetabolic disorder in streptozotocin diabetes mice.CR and BUN index：CR and BUN level in the model and positive group were distinctively different with that in the wild type, indicatingthat total saponins of Stigma Maydis can improve renal function ofthe diabetes mice.Weight, kidney weight, Weight/kidney weight index: The Weight,kidney weight, Weight/kidney weight of model and WT aredistinctively different, indicating that total saponins of StigmaMaydis has a kidney protection by decreasing renomegal.Kidney morphology observation: WT’s glomerular volume was normal,the glomerular capsule was clear. Model mices glomerular wasswelling, and capsule was disformed. Experiment group mice’sglomerular were partly swelling, while glomerular capsule wasnormal, which indicated that total saponins of Stigma Maydis hasa prevention effect to diabetes mice.Conclusion: We studied on extract process of total saponins ofStigma Maydis by using Semi-bionic method at the first time, andcompared with water extract and ethanol extract. Results indicatedthat Semi-bionic method was better than water extract and ethanolextract, providing a basis for industrial production of totalsaponins of Stigma Maydis. Purified total saponins has a preventioneffect to lipid metabolic disorder of diabetes mice and diabeteskidney disease. |
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