2017)Immunomodulatory activity of small molecular (≤3 kDa) Coix glutelin enzymatic hydrolysate, CyTA -Journal of Food, 15:1, 41-48, ABSTRACT In this study, small molecular (≤3 kDa) Coix glutelin hydrolysate (CGH) was obtained by the method of enzymatic hydrolysis and then immunomodulatory activity was evaluated in vitro and in vivo. Stimulation index of mice splenocytes was increased in a dose-dependent manner after treated with 3.125-25 μg/mL CGH. Acid phosphatase in mice peritoneal macrophages was significantly improved after stimulated with CGH at some concentration. The CGH stimulated the secretion of NO from Raw264.7 in a concentration-dependent manner and also significantly inhibited lipopolysaccharideinduced cell excessive activation at some extent. After successive administration of CGH for 21 days, the spleen index of ICR was significantly increased to 3.73 mg/g for the group administrated 800 mg/kg CGH. Malondialdehyde (MDA) assay displayed that 200 mg/kg CGH increased mice serum MDA concentration, while the variation was obsolete in the other CGH groups. All results demonstrated that CGH took essential effects on immunomodulatory activities.Actividad inmunomoduladora de hidrolisato glutelina encimático de moléculas pequeñas de Coix (≤ 3 kDa) RESUMEN En este estudio, se obtuvo hidrolisato glutelina de Coix (CGH) de molécula pequeña (≤ 3 kDa) mediante el método de hidrólisis enzimática y después se evaluó la actividad sinmunomoduladora in vitro y in vivo. El índice de estimulación (SI) de los esplenocitos de ratón aumentó de forma dependiente a la dosis después ser tratados con 3,125-25 μg/mL de CGH. La fosfatasa ácida (ACP) en los macrófagos peritoneales de ratón mejoró significativamente después de que se estimulara con CGH en algunas concentraciones. El CGH estimuló la secreción de NO a partir de Raw264.7 de forma dependiente de la concentración e inhibió de forma significativa la activación excesiva de células inducidas con LPS en cierta medida. Después de una administración sucesiva de CGH durante 21 días, el índice esplénico de ICR aumentó significativamente a 3,73 mg/g para el grupo al cual se había administrado 800 mg/kg de CGH. El ensayo de malondialdehído (MDA) mostró que los 200 mg/kg de CGH hicieron aumentar la concentración de MDA del suero de ratón, mientras que la variación fue obsoleta en los otros grupos de CGH. Todos los resultados demostraron que CGH obtuvo efectos esenciales en las actividades inmunomoduladoras. ARTICLE HISTORY

To analyze the protein composition of Brucea javanica seeds and evaluate the cytotoxicity of its gulbulin hydrolysates. Four protein fractions of albumin, gulbulin, prolamin and glutelin were sequentially extracted and then quantified by Kjeldahl method. Different kinds of proteases were applied to hydrolyze B.javanica gulbulin, and MTT assay was used to evaluate the cytotoxicity of low molecular weight hydrolysates (≤3 kDa) on human breast cancer MCF-7 cell. The results showed that： the total protein content of B.javanica seeds was 17.47%, albumin, gulbulin, coxin, glutelin and residue protein accounted for 15.01%, 8.11%, 2.47%, 44.92% and 23.62% of the total protein content, respectively. The hydrolysates (≤3 kDa) of B.javanica globulin produced by pepsin showed significant growth inhibitory activity on MCF-7 cells, and the IC₅₀ value was（6.52±0.01） mg•L⁻¹ after 72 h of incubation. Protein was abundant in B.javanica seeds, and its peptides demonstrated specific cytotoxicity on MCF-7 cell line in vitro, suggesting antitumor active ingredient can be further generated from B.javanica seeds.