Original scientific paper

Inhibitory effect of the plant proteins mixture on the specific sucrase activity in intestine of diabetic mice

ROBERTA PETLEVSKI ; University of Zagreb, Faculty of Pharmacy and Biochemistry Department of Medical Biochemistry and Haematology, Domagojeva 2, Zagreb, Croatia
MIRKO HADŽIJA ; Ruđer Bošković Institute, Laboratory for Experimental Medicine, Bijenička 52, Zagreb, Croatia
ZLATKO SABOL ; Polyclinic for Children's diseases Dr Sabol, IV. Cvjetno naselje 21, Zagreb

Abstract

Background and Purpose: Increased activity of sucrase, one of the
intestinal alpha-glucosidase founded in diabetes mellitus. Inhibition of sucrase activity, plays a major role in preventing rise in postprandial glucose level in diabetics. On peer-reviewed literature could be found regarding investigation the effect of mixture plant proteins, Mw 3 – 15 kDa (MPP), isolated from Astragali radix – Astragalus membranaceus Fisch., Foenugraeci semen – Trigonella foenum graecum L., Cichorii radix – Cichorium Intybus L. and Urticae radix and herba – Urtica dioica L. on sucrase activity. This plants are used in traditional medicine of treatment of diabetes mellitus. The aim of this study was to determine activity of sucrase in small intestinal homogenates of NOD diabetic mice on feeding with and without MPP in chow.

Materials and Methods: In mice diabetes was induced by i.v. injection
of aloxan-monohydrate (75 mg/kg b.m.) seven days before treatment with MPP. The proteins (Mw 3 – 15 kDa), were isolated from ethanol extract, each plants separately, by gel filtration method on Sephadex G-25 column. Eluted fraction which highest absorbance on 280 nm were pooled, dialyzed, lyophilized and mixed (MPP) and before treatment in mice solvent in sterile PBS. After seven days of treatment diabetic NOD mice with MPP (1,8 g/d), the small intestine was removed and divided into three segments, from pylorus to duodenum, and two equal lengths of the jejunum and ileum and homogenized in cold 0.14M KCl. Specific sucrase activity was determined using method of Dahlquist et. al., by sucrose as substrate.

Results and Conclusion: We confirmed the increased specific sucrase
activity in the intestine of diabetic NOD mice. Our results also indicate that
MPP have strongly inhibitory potential on intestinal sucrase activity
(p<0.05) in diabetic mice. Conclusions drawn from this study should be
further supported and our future experiments will be focused on determining the amino acid sequence of each protein from MPP.

Keywords

diabetes mellitus; plant protein; a-glucosidase; sucrase inhibition

Hrčak ID:

135131

URI

https://hrcak.srce.hr/135131

Publication date:

30.9.2014.

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