Effect And Mechanism Of Sanye Tablet On Blood Glucose For Healthy Subjects After Taking Maltose

Objective:1.To study the effect of Sanye Tablet on blood glucose of healthy people after taking maltose.2.Study on the inhibitory characteristics and types of Sanye Tablet on maltase activity in vitro3.Study on the effect of Sanye Tablet on the secondary conformation of maltase.Methods:1.A two-cycle self-crossover control design was used to select 12 healthy subjects.each subject will receive two kinds of intervention,maltose and maltose+6 pieces of Sanye Tablet,respectively,to observe the effect of three leaves on blood glucose of healthy subjects after taking maltose.2.Using maltose as substrate,the inhibitory effect and mechanism of Sanye Tablet on maltase were studied according to enzyme inhibition kinetics,including:（1）the optimum substrate concentration,reaction time and enzyme concentration were determined first.（2）different concentrations of Sanye Tablet extract reacted with maltase and substrate,and the inhibition rates of different concentrations of Sanye Tablet extract on maltase were calculated,and the half inhibitory concentration(IC₅₀)of Sanye Tablet extract was calculated by curve fitting.（3）the extract concentration and substrate concentration of Sanye Tablet were fixed,and the enzyme quantity-velocity diagram（Emurv diagram）was drawn by changing the amount of enzyme,and the type of inhibitory effect of extract from Sanye Tablet on maltase was judged.（4）fixed the amount of enzyme,changed the substrate concentration,drew the double reciprocal inhibition curve of substrate concentration-reaction rate（[S]-V reciprocal curve）by Lineweave-Burk double reciprocal method,and analyzed the inhibition type of Sanye Tablet on maltase.3.The effects of different concentrations of Sanye Tablet on the molecular conformation of maltase under different reaction time were studied by circular dichroism.Results:1.Clinical study:（1）compared with taking maltose alone,the blood glucose of the subjects after taking maltose+Sanye Tablet decreased,and the blood sugar of 45min,60min and 90min decreased significantly（P<0.05）.（2）the incremental area under the curves of taking maltose was 279.41±111.31,and the AUC of taking maltose+Sanye Tablet was 203.86±61.08,there was significant difference between the two groups（P<0.05）.2.Mechanism studyexperiment one:enzyme inhibition kinetics(1In the selection of the optimal substrate（maltose）concentration,enzyme concentration and time,the maltose concentration 0.25mol/L was determined as the substrate concentration in this experiment,the optimum enzyme reaction concentration was 5mg/m L,and the optimum reaction time was 12min.（2）The half inhibitory concentration(IC₅₀)of extract from three leaves on maltase was（1.64±0.10）×10^-2mg/m L.（3）In the inhibition test,the inhibition curves of trefoil group and acarbose group intersect with the negative control group at the origin,and they are reversible inhibitors.（4）In the experiment of identification of inhibition types,the inhibition curves of different concentrations of Sanye Tablet extract and acarbose intersected on the y-axis with the negative control group,and they were all competitive inhibitors.Experimental two:circle dichroism studyWith the change of the concentration and time of Sanye Tablet,the spectral intensity,peak position and shape of maltase changed,the proportion ofα-helix increased,up to47.90%;the proportion ofβ-folding decreased or even could not be detected;the proportion ofβ-corner increased to 24.20%;the change of irregular crimping was not obvious.Conclusions:1.Sanye Tablet can reduce the blood sugar level of healthy subjects after taking maltose.2.The type of inhibition of Sanye Tablet to maltase was reversible competitive inhibition.3.Sanye Tablet can change the secondary structure of maltase molecules and make maltase lose its natural conformation.4.Sanye Tablet is a potentialα-glycosidase inhibitor.