Echinacea purpurea (L.) Moench root lectin nephrotoxicity

Abstract

Introduction. Lectins or hemagglutinins are non-immune origin glycoproteins, which can bind carbohydrate structures in specific and reversible manner. They have big potential for their therapeutical applications such as immunomodulatory [1], anticancer [2], antibacterial [3] and other activities. Hemagglutinins from Echinacea purpurea (L.) Moench roots haven’t been investigated and tested on animal models in vivo. Aim of experiment: To purify hemagglutinins from Echinacea purpurea (L.) (Moench) roots and test their activity on mice kidney morphology in vivo. Experiment tasks: 1. Purify and identify hemagglutinins from purple coneflower roots; 2. Test hemagglutinin impact on mice kidney morphology in vivo. Materials and methods. 1. Affinity column with immobilized D-(+)-mannose ligands was equilibrated with phosphate buffer saline (PBS) pH 7.4. Unbound proteins were washed out with PBS at the elution rate 1 mL/min. Hemagglutinins were eluted out of the column with 0.2 M lactose solution in PBS. Following hemagglutinin fraction was collected and checked for hemagglutinating activity [4]. Hemagglutinating active fraction was separated by polyacrylamide gel electrophoresis and immunoblotted for glycosylated proteins. Glycosylated protein bands were identified with liquid chromatography tandem mass spectrometry by Echinacea transcriptome and sunflower genome database search. 2. Purified hemagglutinin fractions tested on animal models in vivo (Approved by State Food and Veterinary Service, permission No. G. 2-56). Three different test animal groups were selected: animals (n=15) in negative control group got 50 μL peritoneal injections of physiological NaCl solution, animals in tincture group (n=15) got 50 μL peritoneal injection of Echinacea purpurea (L.) Moench root tincture (1 μl of tincture ... [...]