| Firstly,the inhibition effects of potentilla fruticosa extraction on 3 kinds of enzymes were researched through the extracorporeal screening model of prune-amylase inhibitor?AMI?,post-processing enzyme inhibitor?AGI?and aldose reductase inhibitor?ARI?in vitro.Secondly,the effects of Potentilla fruticosa extraction on the maximum tolerance and postprandial blood glucose of normal little mice were measured,and the effects of potentilla fruticosa extraction on fasting serum insulin?INS?,glucagon?GC?,leptin?LEP?and c-reactive protein?CRP?of big mice suffering from T2DM in vivo were also measured.Finally,the related enzymes expression quantity of insulin?INS?,resistance element?Resistin?,leptin?LEP?and sugar metabolism of big mice suffering from T2DM in vivo were determined and analyzed with fluorescence quantitative rt-pcr technique and Livak method,in addition,the resistance mechanism of potentilla fruticosa extraction to T2DM was expored.1.Water extract?WE?was obtained by soaking with 10 times distilled water,then the different polar parts of potentilla fruticosa were extracted by methyl alcohol back flow extraction,petroleum ether extraction?MEP?and ethyl acetate extraction?MEE?.2.AMI,AGI and ARI screening model In vitro tested that WE and MEE showed the highest inhibitory activity in the three enzyme inhibition experiments,and half of the inhibiting concentration(IC50)of WE and MEE on AM was 0.432 and0.507mg/mL respectively.IC500 of WE and MEE on AG was 0.164 and 0.466mg/mL.IC500 of WE and MEE on AR were 0.742,1.098 mg/mL.the results of the inhibition activity of 11 monomer compounds to enzyme suggested that the inhibition activity the quercetin 3-O-?-D-glucuronide?3?and 1,6-di-O-galloyl-?-D-glucose?6?to AG were much better than others,and the IC500 were 0.058 and 0.362mg/mL.3.The maximum tolerance results showed that the maximum tolerance of WE and MEE to mice were 18.75g·Kg-1/d and 16.5·Kg-1/d,respectively.the blood sugar value of mice with starch load after giving WE and MEE after 30,60,90,90 min in high dose group was reduced?p<0.05,p<0.01?,the blood sugar value of mice with starch load after giving WE and MEE after 30,60 min in low dose groupwas was also reduced?p<0.05,p<0.01?.4.After WE high,medium and low dose gave T2DM rats,the three polys and one little symptoms were improved.The results of blood glucose observation showed that at the 45d,53d and 60d of administration,there was a significant difference between the WE high dose group and the model group?p<0.05?.At the 53d and 60d of administration,there was a significant difference between the WE medium dose group and the model group?p<0.05?.At the 60d of administration,there was a significant difference between the WE low dose group and the model group?p<0.05?.5.In T2DM rats treated with water extract of potentilla fruticosa?WE?,the content of INS was higher than that in the model group,and GC,LEP and CRP were lower than those in the model group.6.Compared with the T2DM rat model group,the mRNA expression of Resistin,LEP and G6pase genes in the WE high dose group significantly decreased?p<0.01?,and the mRNA expression of INS and GK genes significantly increased?p<0.05?.The mRNA expression of Resistin,LEP and G6pase genes in rats in the WE medium dose group was significantly decreased?p<0.05,p<0.01,p<0.05?,and the mRNA expression of INS and GK genes was significantly increased?p<0.05?.The mRNA expression of Resistin,LEP and G6pase genes in rats in the low dose group decreased significantly?p<0.01,p<0.01,p<0.05?,and the mRNA expression of INS and GK genes increased significantly?p<0.05?.The results show that potentilla fruticosa can promote the mutual adjustment between insulin and glucagon,could improve the oral glucose tolerance test in mice,can promote insulin secretion,improve insulin sensitivity,have fall blood sugar effect. |
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