Investigation of the transport and absorption of Angelica sinensis polysaccharide through gastrointestinal tract both in vitro and in vivo

Abstract

To investigate the absorption and delivery of ASP in gastrointestinal (GI) tract, cASP was successfully synthesized by chemically modifying with succinic anhydride and then conjugating with a near infrared fluorescent dye Cy5.5. Then, the capacity of oral absorption of cASP was evaluated. The results demonstrated that cASP had low toxicity and no disruption on the integrity of cell membrane. The endocytosis of cASP into the epithelial cells was time- and energy-dependent, which was mediated by macropinocytosis pathway and clathrin- and caveolae (or lipid raft)-related routes. Otherwise, the actin filaments played a relatively weak role at the same time. The transport study illustrated that cASP could penetrate through the epithelial monolayer and mainly mediated by the same routes as that in the endocytosis experiment. Moreover, both in vitro Ussing chamber and in vivo ligated intestinal loops models indicated that cASP could be diffused through the mucus barriers and be absorbed in the whole small intestine. Finally, near-infrared fluorescence imaging presented that cASP could be absorbed and circulated into the blood, then distributed into various organs after oral administration. In conclusion, ASP could be absorbed after oral administration through endocytosis process mainly mediated by macropinocytosis pathway and clathrin- and caveolae (or lipid raft)-related routes, then be absorbed and circulated into blood. This study presents a comprehensive understanding of oral delivery of cASP, which will provide theoretical basis for the clinical application of ASP.

Anti-Inflammatory and Hepatoprotective Effects of <b><i>Ganoderma lucidum</i></b> Polysaccharides against Carbon Tetrachloride-Induced Liver Injury in Kunming Mice

“…lucidum, a Chinese medicinal mushroom, has been widely used in the treatment of chronic hepatopathy with little or no side effects of various etiologies [15]. It could be absorbed into the epithelial cells after oral administration through endocytosis process mainly mediated by macropinocytosis pathway and clathrin -and caveolae (or lipid raft) -related routes and then be absorbed and circulated into blood [16]. Polysaccharide in G. lucidum (GLPS) has been proposed as the bioactive constituents responsible for the protective activities against toxin-induced liver injury [15].…”

Immunomodulatory Effects of Lycium barbarum Polysaccharide Extract and Its Uptake Behaviors at the Cellular Level

“…The human colon carcinoma cell line, Caco-2 could slowly differentiate to form a cell monolayer that possesses many functions of the small intestinal villus epithelium. Caco-2 cell monolayer has been widely used in the early screening of drug development and had become an important model to study the mechanism of drug absorption and transport [24]. The above results showed that LBP had no significant effect on the viability of Caco-2 cells, but it could be internalized into Caco-2 cells and partially locate in lysosomes ( Figure S11), suggesting no toxicity of LBP to Caco-2 cells.…”

“…Due to the large molecular mass and high hydrophilicity of polysaccharides, it is difficult to enter cells by passive diffusion [39]. Some studies have shown that clathrin mediates the internalization process of polysaccharides [24]. The clathrin-dependent pathway includes three key steps: cell membrane infolding that depends on clathrin, vesicle formation that relies on dynamin, and endosome maturation.…”

“…As hydrophilic macromolecules, whether polysaccharides can be absorbed by oral administration is a controversial issue [24]. At present, oral administration is the only way to take LBP.…”

“…Because of the structural heterogeneity and the lack of chromophore, the quantitative study of the uptake behavior of polysaccharides by gastrointestinal tract and cells is very difficult. Fluorescence-based bioimaging technology has been widely used in the field of bioimaging because of its inherent high sensitivity, high selectivity, convenience and non-invasiveness [25], and has been used to track cellular uptake and endocytosis of polysaccharides [24,26]. Caco-2 cells are derived from human colonic adenocarcinoma cells and can undergo epithelial differentiation to form a single cell layer with similar structure and function to the intestinal epithelium [27].…”