Abstract
The monocot lectin from the tubers of Arisaema erubescens (Wall.) Schott has been purified by consecutive hydrophobic chromatography and ion exchange chromatography methods. The molecular weight of this A. erubescens lectin (AEL) was determined to be about 12 kDa by high performance liquid chromatography (HPLC) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) methods. AEL could agglutinate rabbit erythrocytes. The haemagglutination activity of AEL was only inhibited by asialofetuin, while monosaccharide did not react. Rat paw edema and neutrophil migration models were used to investigate the pro-inflammatory activity of AEL. AEL (100 and 200 μg/paw) could induce significant rat paw edema. In addition, AEL (100, 200 and 300 μg/mL/cavity) could induce significant and dose-dependent neutrophil migration in the rat peritoneal cavities. Besides, AEL at doses ranging from 100 to 300 μg/mL/cavity could significantly increase the concentration of nitric oxide (NO), prostaglandin E2 (PGE2) and tumor necrosis factor alpha (TNF-α) in peritoneal fluid. As compared with control animals, 75% depletion in the number of resident cells following peritoneal lavage did not reduce the AEL-induced neutrophil migration. However, pre-treatment with 3% thioglycollate which increased the peritoneal macrophage population by 201%, enhanced the neutrophil migration induced by AEL (200 μg/mL/cavity) (p < 0.05). Reduction of peritoneal mast cell population by chronic treatment of rat peritoneal cavities with compound 48/80 (N-methyl-p-methoxyphenethylamine with formaldehyde) did not modify AEL-induced neutrophil migration. The results provided the basis for identifying the toxic components of A. erubescens and AEL could be a new useful tool for pro-inflammatory research.
Highlights
Arisaematis Rhizoma (AR) is the rhizomes of Arisaema erubescens (Wall.) Schott, which has been widely used in Traditional Chinese Medicine for thousands of years
The main peak which was obtained by hydrophobic interaction chromatography was eluted with a linear gradient of 0–0.4 mol/L NaCl at a flow rate of 1 mL/min
It is possible that A. erubescens lectin (AEL) does not directly activate mast cells, but could be releasing inhibitory neutrophil chemotactic factors. These findings suggested that AEL-induced neutrophil migration follows an indirect pathway and may be dependent on the release of neutrophil chemotactic factors from resident macrophages
Summary
Arisaematis Rhizoma (AR) is the rhizomes of Arisaema erubescens (Wall.) Schott, which has been widely used in Traditional Chinese Medicine for thousands of years. Our previous research had proved that AR demonstrated toxicity due to the components of raphide including calcium oxalate, protein and trace carbohydrates [3,4]. Investigations showed that the toxicity of the raphide might closely relate to the protein components [5]. Lectin from Vatairea macrocarpa could induce paw edema in rats [12]. It could induce neutrophil migration in vivo [13]. It was deemed necessary to investigate the toxic components of AR in order to ensure a more safe and effective use in clinical treatment. The pro-inflammatory effects of AEL were evaluated by rat paw edema and neutrophil migration into rat peritoneal cavity models. The relationship of AEL-induced neutrophil migration and the possible involvement of resident cells, macrophages as well as mast cells were investigated
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