Biological Mechanisms Induced by Soybean Agglutinin Using an Intestinal Cell Model of Monogastric Animals

Soybean agglutinin (SBA) has a toxic effect on most animals. The anti-nutritional mechanisms of SBA are not fully understood, in terms of cell survival activity and metabolism of intestinal cells. This study aims to investigate the effects of SBA on the cell cycle, apoptosis, and to verify the mechanism of SBA anti-nutritional characters based on proteomic-based analysis. The IPEC-J2 cell line was cultured with medium containing 0.0, 0.5, or 2.0 mg/mL SBA. With increasing SBA levels, the percentage of the cells at G0/G1 phase, cell apoptosis rates, expressions of Bax and p21, and the activities of Casp-3 and Casp-9 were increased, while cyclin D1 and Bcl-2 expressions were declined (p < 0.05). The proteomic analysis showed that the numbers of differentially expressed proteins, induced by SBA, were mainly enriched in different pathways including DNA replication, base excision repair, nucleus excision repair, mismatch repair, amide and peptide biosynthesis, ubiquitin-mediated proteolysis, as well as structures and functions of mitochondria and ribosome. In conclusion, the anti-nutritional mechanism of SBA is a complex cellular process. Such process including DNA related activities; protein synthesis and metabolism; signal-conducting relation; as well as subcellular structure and function. This study provides comprehensive information to understand the toxic mechanism of SBA in monogastrics.

Vapor-Stripping and Encapsulating to Construct Particles with Time-Controlled Asymmetry and Anisotropy

An innovative chemical vapor sublimation and deposition (CVSD) process was shown to produce nanoscale anisotropic hybrid materials. Taking advantage of controlled thermodynamic properties and the mass transfer of molecules, this process allowed for water vapor sublimation from an iced template/substrate and stagewise vapor deposition of poly-p-xylylene onto the sublimating ice substrate. In this study, the use of sensitive soybean agglutinin (SBA) protein tubes was demonstrated as an example to prepare the anisotropic hybrid material based on the CVSD process. The rationale of a timing parameter, Δt, was controlled to program the sublimation of the SBA-ice templates and the deposition of poly-p-xylylene during the CVSD process. As a result of this control, a stripping stage occurred, during which SBA tubes were exposed on the particle surface, and a subsequent encapsulation stage enabled the transformation of the ice templates into a nanometer-sized anisotropic hybrid material of poly-p-xylylene as the matrix with encapsulated SBA tubes. The timing parameter Δt and the controlled stripping and encapsulating stages during CVSD represent a straightforward and intriguing mechanism stemming from physical chemistry fundamentals for the fabrication of hybrid materials from sensitive molecules and with predetermined sizes and asymmetrical shapes. A simulation analysis showed consistency with the experimental results and controllability of the timing mechanism with predictable particle sizes.

The Whole Cell Proteome-based Study on The Mechanism of Apoptosis Induced by Soybean Agglutinin on IPEC-J2

Background: Soybean agglutinin (SBA), a major anti-nutritional factor in soybean, may induce abnormal health and metabolism of intestinal cells, resulting in the reduction of the production performance of animals. The anti-nutritional mechanisms of SBA are not fully understood, in terms of the cell life activities and metabolism of intestinal cells. This research aims to find the effects of SBA on the cell cycle, apoptosis and proteomic, and furtherly to get more findings for verifying the mechanism of SBA anti-nutritional characters.Methods: The IPEC-J2 cell line was cultured with the medium containing 0.0, 0.5 or 2.0 mg/mL SBA, respectively, for 24 h. The percentage of the cells at different cell cycle phases (G0/G1 phase, S phase and G2 phase) and cell apoptosis rates were measured with flow cytometry. The expressions of Cyclin D1, active p21, Bcl-2, and Bax were determined by western blotting. The activity of caspase-3 (Casp-3) and caspase-9 (Casp-9) were tested with ELISA. The whole-cell quantitative proteome were detected by TMT/iTRAQ Labeling, HPLC fractionation and LC-MS/MS Analysis. The functions and characteristics of the differential expressed proteins in the proteome results were analyzed from the aspects of GO annotation and KEGG pathway. The relationship between the results of proteomics and apoptosis or cell cycle were analyzed and discussed.Results: The percentage of the cells at G0/G1 phase, cell apoptosis rates, expressions of Bax and p21, and the activities of Casp-3 and Casp-9 were increased, cyclin D1 and Bcl-2 expression were declined with the increased of the SBA treatment levels (p<0.05). The proteomic measurements showed that a numbers of differentially expressed proteins, caused by SBA treatment, were mainly enriched in the DNA replication, base excision repair, nucleus excision repair, mismatch repair, ubiquitin-mediated proteolysis pathway, cell structural-proteins, and structures and functions of mitochondria. Moreover, the differential expressed proteins enriched in AMP-activated protein kinase (AMPK) pathway and the process of synthesis and metabolism of proteins were only found in 2.0 mg/mL SBA treatment.Conclusion: The results of this experiment demonstrated that cell cycle arrest and apoptosis induced by SBA may be resulted from the downregulating the expression of the proteins related to DNA replication and repair, protein translation, signal-conducting relation, cell structure, and subcellular structure and function.

FEATURES OF DISTRIBUTION OF SOYBEAN AGGLUTININ (SBA) RECEPTORS IN THE EXTRACELLULAR MATRIX OF THE MENISCI OF RAT KNEE JOINT AFTER INTRAFETAL INJECTION OF ANTIGENS

FEATURES OF DISTRIBUTION OF SOYBEAN AGGLUTININ (SBA) RECEPTORS IN THE EXTRACELLULAR MATRIX OF THE MENISCI OF RAT KNEE JOINT AFTER INTRAFETAL INJECTION OF ANTIGENS