Studies On The Inhibitory Effects Of Active Component From TCM On α-glucosidase

a-glucosidase can be find in a variety of organisms. They are not only the critical enzyme for the body’ metabolism but also relevant to a lot of diseases. Using a-glucosidase as a target, screening a-glucosidase inhibitors from traditional Chinese medicine (TCM) provides a huge boost to the exploitation of new medicine and the modernization of TCM. In this paper, two experiments were explored to solve the shortcomings in the screening.A novel screening method for a-glucosidase inhibitors has been establish by HPLC in vitro. The enzyme activity of a-glucosidase were determined by monitoring the release of PNP from the substrate PNPG The optional conditions for the enzyme-catalyzed reaction by single factor test was developed as follows:the reaction mixture contained 10 μL of 0.067 mol·L-1 phosphate buffer (pH 6.8) or the indicated concentration of sample solution,30μL of 0.1 U·mL-1 a-glucosidase, shaked until homogeneous, the reaction was processed at incubated at 37℃ for 20 min and added 40 μL of 8.0 mmol·L-1 PNPG, shaked again, the mixture was incubated at 37 ℃ for 30 min and stoped by adding 80 μL of 0.2 mol·L-1 Na2CO3, then diluted by adding 340 μL water, shaked again and filered by 0.45 μm of membrane before detected by HPLC. The inhibitory rates (%) were calculated according to the change of concentration of PNP. Acarbose was used to validate the established method. This methed was marginal, simple and sensitive with quick effectiveness which could reduce the false positive proportion of the screening and increase the odds of searching new compounds from complex mixtures or pigment jamming samples.Fifty species of TCM which have remedial effect to diabetes mellitus traditionally were investigated in this paper. These traditional Chinese medicine were systematic extracted by n-hexane, methylene dichloride, ethylacetate, methanel and aqueous. So a multi-component library was built up which contained 250 species of compounds, and the inhibitory activity of these extracts against a-glucosidase under tree different concentrations (64,6.4,0.64 mg·mL-1) were analysised by HPLC method. This was the first time that screening a-glucosidase inhibitors from TCM by HPLC. The TCM and its extracts which have the higher inhibition activity were determined by the screening, and the characers and laws of TCM were concluded. This screening provided important information in the exploitation and utilization of to find higher and newer inhibitors TCM against a-glucosidase.