The extract of Seabuckthorn is a Chinese medicine for allergic or inflammatory diseases (including obstructive pulmonary diseases). We have demonstrated the activation of a 36kD MBP kinase, Mst1, during Fas-activating antibody-induced apoptosis of human CD4+ T-cells, as well as human eosinophils. In this study, we determined whether the extracts of Seabuckthorn seeds (SS) and the subfractions (S1, S2, S3, S4, S5 and S6) promote apoptosis, and activate MBP kinase in Jurkat and HL-60 cells. Both Jurkat and HL-60 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum. After 24 h of cell arrest, the cells were treated with the extracts. Apoptotic cells were quantified using Annexin V-FITC/PI, and Western blotting and "In-Gel" assay were performed on the lysate. SS and S3 (10 – 100 μg/ml) induced apoptosis of both Jurkat and HL-60 cells, and also activated the 36kD MBP kinase in the cells. Western immunoblot analysis showed that the 36kD molecule bound with anti-Mst1 antibodies. S3-induced apoptosis of HL-60 was significantly greater than that of Jurkat. These results suggest that the extracts of Seabuckthorn seeds-induced apoptosis is at least partly associated with Mst1 in both Jurkat and HL-60 cells.
This abstract is funded by: This work is supported by a grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan to promote multi-disciplinary research projects.