Inhibitory Activity And Hypoglycemic Effect Of Actinidia Chinensis Planch Roots On ?-glucosidase

Diabetes is a metabolic disease caused by absolute or relative deficiency of insulin secretion,the various complications it causes have become major public health problems affecting human health.Traditional Chinese medicine has a long history of treating diabetes in China,and it shows good curative effect.Therefore,it is hot to select the hypoglycemic drugs with good curative effect from Chinese herbal medicines.Actinidia chinensis Planch is a common ethnic medicinal material in Guizhou.Studies have shown that Actinidia chinensis Planch roots has anti-inflammatory,anti-tumor,anti-oxidation,hypolipidemicandother pharmacological effects,it has a strong inhibitory effect on?-glucosidase.Therefore,this paper is devoted to screen natural?-glucosidase inhibitors from the Actinidia chinensis Planch roots,in order to find new safe and effective drugs.The main research contents are as follows:1.In this paper,3kg of Actinidia chinensis Planch roots was extracted by heating with 75%ethanol,the extract was extracted with petroleum ether,ethyl acetate and n-butanol in that order.At a concentration of 100?g/ml,the inhibition rate of Actinidia chinensis Planch roots was 96.77%by in vitro activity inhibition test,and the competitive inhibition of?-glucosidase by Actinidia chinensis Planch roots was determined to be competitive inhibition by Lineweave-Burk double reciprocal method.The inhibition rates of the three extraction sites were 86.62%?74.91%?96.55%.Among them,the inhibitory activity of the n-butanol extraction site was the most obvious,and the IC₅₀0 was 3.12?g/mL.2.In order to further screen the monomeric compound which has strong activity,the n-butanol fraction was obtained by silica gel column chromatography and preparative thin layer chromatography to obtain two compounds,which were identified by chromatographic method and literature comparison as compound I??-Sitosterol?,compound II?catechin?,At a concentration of 100?g/ml,the IC₅₀0 of the two was 66.88?g/mL and 19.33?g/mL.3.Screening of?-glucosidase inhibitors in Actinidia chinensis Planch roots by affinity ultrafiltration combined with liquid chromatography?UF-LC-MS?:Seven compounds were screened by affinity ultrafiltration.Combined with LC-MS technical literature reference,three compounds were identified,namely isozyme,?-sitosterol and chlorogenic acid,and the structure of the remaining four compounds was to be determined.The three compounds analyzed were docked online by the Systemsdock method,and the Docking scores were 7.228?4.992?4.918.This further demonstrates that all three compounds have good binding activity to?-glucosidase.4.The toxic and glucose consumption of kiwi root and its active site on HepG2cells:MTT assay showed that the medium and low concentrations of kiwi root and n-butanol were not toxic to HepG2 cells,high concentration with low toxicity;glucose consumption test results indicated:after the total extract of kiwi root and its n-butanol extract acted on HepG2 cells,the glucose content of the cell supernatant decreased,which proved that Actinidia chinensis Planch roots total extract and n-butanol extract could promote the glucose consumption of normal HepG2 cells.The n-butanol site has a tendency to promote insulin resistance to HepG2 cells.In summary,this paper screens the active sites and reactive monomer compounds that produce?-glucosidase inhibitory activity in the roots of kiwi by using traditional separation methods and rapid efficient method of UF-LC-MS.Vitro activity assay,Molecular docking method,HepG2 cell experiment demonstrated that the extract of kiwi root n-butanol and several monomeric compounds can inhibit the activity of?-glucosidase and promote the glucose consumption of HepG2 cells.Reveal the material basis of hypoglycemic action and mechanism of kiwi roots.