Study On The Effect And Mechanism Of Three Plant Active Components On Oxidative Stress In Type 2 Diabetic Mice

Diabetes mellitus is a chronic metabolic disease caused by a variety of factors.Its etiology and pathogenesis are not fully clear,and a lot of studies have shown that it is closely related to oxidative stress.At present,most of the drugs used in diabetes are western medicine,which may bring some side effects for a long time.Pumpkin polysaccharide,puerarin flavonoids,Pu-erh thearubigins extracted from plants is reported to have hypoglycemic,lipid-lowering,antioxidant,anti-inflammatory and other effects,which side effects are small,safe and cheap,causing more and more scholars attention.People have in-depth study on the physiological activity and its effect on the active components of the three kinds of plants,but the research of combined effect in improving type II diabetes is little.And a variety of plant active ingredients used in combination are characterized by multi level and multi targets.Therefore,the type 2 diabetes model was established to analyze the effect and mechanism of pumpkin polysaccharide,pueraria flavonoids and pu-erh theabrowns on glycolipid metabolism and oxidative stress by single and combined use.First of all,the type 2 diabetes model was established to explore the improvement of the three kinds of plant extracts on glycolipid metabolism by single and combined use;Secondly,to explore the effect of single and combined use of the three kinds of plant extracts on oxidative stress,and analyze the relationship of glycolipid metabolism and oxidative stress.Principal component analysis(PCA)was used to obtain the principal components of different indexes,and compare the differences of target points between single and combined groups to explain its potential mechanism.Finally,we used the Western blot method to study the expression of the oxidative stress pathway Nrf2/HO-1 in the single and combination groups.The mechanism of action was analyzed from the view of molecular biology.The main results are as follows:(1)pumpkin polysaccharide,pueraria flavonoids and pu-erh theabrowns extracts single and combined use can effectively improve the typical symptoms and the disorders of glycolipid metabolism in type 2 diabetes mice,and the combination group have more obvious effect than the single group.In the area under blood glucose(AUG),fasting blood glucose(FBG),insulin(INS),total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),the effect of pu-erh theabrowns and pueraria flavonoids combination group was significantly better than pueraria flavonoids group(P<0.05);In the INS 、 TG,the effect of pu-erh theabrowns and pueraria flavonoids combination group was significantly better than that of theabrownine group(P<0.05);the effect of pumpkin polysaccharide and pu-erh theabrownscombination group and pumpkin polysaccharide and pueraria flavonoids group is significantly better than pumpkin polysaccharide group in TG(P<0.05).The correlation analysis showed that there was a significant linear correlation between the glucose and lipid metabolism index(P<0.01).We can speculate that the three extracts may improve insulin resistance,regulating dyslipidemia,so as to form a good circulation of glucose and lipid metabolism,slow metabolism disorder in type 2 diabetes,and the combined use of the three kinds of extracts have synergistic effects through multiple targets.(2)pumpkin polysaccharide,pueraria flavonoids and pu-erh theabrowns extracts single and combined use have significant effects on oxidative stress in type 2 diabetic mice.In addition to pumpkin polysaccharide and pu-erh theabrowns combination group in free fatty acids(FFA)and malondialdehyde(MDA),other extracts combined use have better effect than the single group.The effect of pumpkin polysaccharide and pueraria flavonoids combination group was significantly better than the single groups in FFA、MDA(P<0.05).The effect of pueraria flavonoids and pu-erh theabrowns combination group was significantly better than the pueraria flavonoids group in FFA(P<0.05).The results indicated that the combined use of pumpkin polysaccharide,pueraria flavonoids and pu-erh theabrowns had a synergistic effect on oxidative stress.The correlation analysis of the three kinds of extracts in insulin resistance,glycolipid metabolize,oxidative stress showed that oxidative stress is closely related to glycolipid metabolism and insulin secretion.The principal component analysis explain tentatively the potential mechanism of improved type 2 diabetes using single and combined extracts and its changes in the targets and approaches.We speculated that thebetter effect of extracts combined use may be associated with an improvement in the regulation of blood lipids and insulin.(3)pumpkin polysaccharide,pueraria flavonoids and pu-erh theabrowns extracts single and combined use can increase the Nrf2 and HO-1 protein in the liver and colon,and the effect was significant berrer than the model group(P<0.05).It can be seen that the three extracts single and combined use can enhance the expression of Nrf2 protein and its downstream antioxidant protein HO-1,so as to play its antioxidant effect better,improve insulin resistance and protect the body’s normal metabolism.We also found that the expression of Nrf2 and HO-1 in the combined group was stronger than the single group.In the livers of mice,the expression of Nrf2 and HO-1 protein has significant difference between pumpkin polysaccharide and pueraria flavonoids combination group and polysaccharide group(P<0.05),pueraria flavonoids and pu-erh theabrowns combination group were significant differences in pu-erh theabrowns(P<0.05).The expression of Nrf2 protein inmice colon,there is significant difference between pumpkin polysaccharide and pueraria flavonoids combination group with its single group(P<0.05),pueraria flavonoids and pu-erh theabrowns combination group were significant differences in pu-erh theabrowns(P<0.05);HO-1 protein expression differences between pumpkin polysaccharide and pueraria flavonoids combination group and polysaccharide group(P<0.05),tea + ketone group and tea brown differences between treated group(P<0.05).It indicated that the three extracts of combination may be synergistic through multiple targets in the activation of the Nrf2/HO-1pathway.