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KMID : 0861020130280040007
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Korea Journal of Herbology
2013 Volume.28 No. 4 p.7 ~ p.16
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Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells
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김근회:Kim Kun-Hoae
조인식:Cho In-Sik/김한영:Kim Han-Young/김승형:Kim Seung-Hyung/김동선:Kim Dong-Seon/이영철:Lee Young-Cheol
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Abstract
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Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-1+ hematopoietic stem cells (Sca-1+ HSCs) as a testing system.
Methods : Sca-1+ HSCs isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of CD117+ /Sca-1+ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed.
Result : When Sca-1+ HSCs were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-1+ HSCs. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-1+ HSCs. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of ranulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro.
Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/ STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.
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KEYWORD
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Hematopoietic, Brassica campestris var narinosa, Canavalia gladiata DC semen, Cyclophosphamide, JAK2, GATA-1, STAT-5a/b
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