The Study Of Hypoglycemic Material Base Of Picris Japoncia Thunb.

Picris japonica Thunb. belonged to Compositae family. It was often used with heaba and had clear-heat, detumescence and analgesic effect. In Chinese community. Picris japonica Thunb. was devoted to prevente and cured diabetes. The early study of our group showed that the aqueous extract of Picris japoncia Thunb. possessed significant hypoglycemic activitie on diabetic mice. In this paper, we studied the hypoglycemic material base of Picris japoncia Thunb.. The capital contents of this article were summarized as follows:searching for the hypoglycemic utility part of Picris japoncia Thunb.; to be sure the preparation and stabitily of total flavonoids of Picris japoncia Thunb.; to be sure hypoglycemic activity, pharmacodynamics and mechanism of total flavonoids of Picris japoncia Thunb.1.To determine the hypoglycemic effect part of Picris japoncia Thunb.In this part, we did such studies as follows:first, the different of hypoglycemic effects between water extract and 95% alcohol extract of Picris japonica Thunb.:second, the different of hypoglycemic effects and chemical constituent among different parts of water extract of Picris japonica Thunb.. third, the hypoglycemic and hypolipidemic effect of the partⅠof Picris japonica Thunb..The results indicated that, Compared with the diabete model group, the low dose of part I of Picris japonica Thunb. could lowered alloxan-induced diabetic mice's glucose level while the high dose lowered epinephrine-induced diabetic mice's glucose level obviously, and reduced TG, LDL-C content and enhanced HDL-C content. The partⅠof Picris japonica Thunb. had copious flavonoids and polysaccharides. All of above indicated that the partⅠwas the hypoglycemic effect part of Picris japoncia Thunb. while it could regulate blood fat.2. To be sure the preparation of total flavonoids of Picris japoncia Thunb.The extraction technology of total flavonoids was optimized by orthogonal experiment with the content of total flavonoids as index and with ethanol concentration, the ratio of material to solvent, extracting time and extracting frequency, extracting temperature as factors. The optimal extraction technology was that as follow:the concentration of ethanol was 40%; extracting frequency was 2 times, the first extracting time was 90 min and the second extracting time was 60 min; the ratio of material to solvent was 1:20, the extracting temperature was 70℃The macroporous adsorptive resins were taken to purify the active units of Picris japoncia Thunb. We investigated the resin types,the concentration of ethanol precipitation,the water dose and the concentration of ethanol of gradient elution to determine the best purification techniques:D101 type macroporous adsorptive resins,used 70% ethanol to precipitate polysaccharides,the water dose was 1.6 column volume and the concentration of ethanol was 50% of gradient elution. The purity of total flavonoids was ranged from 50% to 53%.The flavonoids from 4 samples of Picris japoncia Thunb. were analysized with HPLC using a step gradient of acetonitrile-0.1% phosphoric acid. The HPLC fingerprint chromatogram of flavonoids from Picris japoncia Thunb. was established using computer software to evaluate the data of chromatographic peaks. It was effective to evaluate the quality of the flavonoids from 4 samples of Picris japoncia Thunb. in virtue of the characteristic peaks of HPLC fingerprint chromatogram. The results indicated that the yield, purity and fingerprint chromatogram of the flavonoids from 4 samples of Picris japoncia Thunb. possessed high semblance and RSD<3% which established material basis of pharmaco-experiment. The semblance of fingerprint chromatogram of the flavonoids from 4 samples of Picris japoncia Thunb. was 92%.3. To be sure hypoglycemic activity of total flavonoids of Picris japoncia Thunb.Impacts of the flavonoids of Picris japoncia Thunb. on alloxan diabetes and adnephrin diabetes mices were observed, while acute toxicity tests were carried out by the largest dosage-concentration and the largest dosage-volume. The results indicated that the flavonoids of Picris japoncia Thunb. could lower the blood glucose level of alloxan diabetes mice and adnephrin diabetes mice with little toxicity and side effect.4. To investigate the hypoglycemic-pharmacodynamics and mechanism of the flavonoids of Picris japoncia Thunb.Diabetes rat model was established by using streptozotocin and biochemical method was applied to examine the influences of Picris japonica Thunb on blood glucose, blood fat. body weight, liver weight, kidney weight, SOD, MDA, BUN of diabetes rat. The HE staining was applied to observe the morphology of pancreatic and kidney, while the immunohistochemisty method was applied to observe the expression of TNF-αand CDlla on kidney. The results revealed that, compared with the diabete model group, the flavonoids of Picris japoncia Thunb. lowered STZ-induced diabetic rat glucose level obviously (p<0.05). while it was better than Picris japonica Thunb aqueous extract and partⅠ(p<0.05) and had no significant difference compared with glibenclamide (p>0.05); hydroposia amount and ingestion of diabetes reduced while their weight aroused; it lowered the content of low density lipoprotein and increased the content of high density lipoprotein significantly.(p<0.05) and better than Picris japonica Thunb aqueous extract, partⅠand glibenclamide(p<0.05); it enhanced SOD-vigor and lowered MDA-content and had no significant difference compared with glibenclamide (p>0.05); it also lowered BUN-content(p<0.05). It had protective effect on pancreatic and kidney and lowered the expression of TNF-a and CD11a on kidney.The reducing prower of the flavonoids of Picris japoncia Thunb. was determined by using the System of K3[Fe(CN)6] and TCA; the activities of scavenging-OH radical by using the system of 1.10-Phenanthroline monohydrate. The results showd that the flavonoids of Picris japoncia Thunb. had the capability of deoxidizing themselves and eliminating the·OH free radical. Moreover, all of above had dose-effect relationship...