人参总皂甙对人GM-CSF和GM-CSFR表达的调控机制

王莎莉1, 陈笛1, 王亚平2, 刘永刚2, 姜蓉2

重庆医科大学基础医学院 1生理教研室, 2组织胚胎学教研室, 重庆 400016

摘要: 为探讨人参调控粒细胞发生的生物学机理, 采用造血祖细胞、 骨髓基质细胞体外培养、 造血生长因子生物学活性检测、 免疫细胞化学、 核酸分子原位杂交、 免疫沉淀和蛋白印迹等现代生物学技术, 研究人参总皂甙(total saponins of Panax Ginseng, TSPG)对人粒-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor, GM-CSF)和粒-巨噬细胞集落刺激因子受体α (GM-CSFRα)表达的影响。结果: (1) 经TSPG (50 μg/ml)诱导制备的骨髓基质细胞、 胸腺细胞、 脾细胞、 血管内皮细胞和单核细胞条件培养液可显著提高粒单系造血祖细胞(CFU-GM)的集落产率; (2) 经TSPG (50 μg/ml)诱导后, 上述细胞的GM-CSF蛋白(诱导24 h)和mRNA(诱导12 h)表达显著提高; (3) 经TSPG (50 μg/ml)诱导24 h骨髓造血细胞的GM-CSFRα蛋白表达增强; (4) 经TSPG (50 μg/ml)刺激后2 min, GM-CSFRα和Shc发生酪氨酸磷酸化, 5 min时达高峰, 随后去磷酸化。上述结果表明, TSPG可能通过直接和/或间接途径促进淋巴细胞与骨髓基质细胞合成与分泌GM-CSF, 诱导骨髓造血细胞表达GM-CSFRα, 并刺激GM-CSFRα和Shc的酪氨酸可逆磷酸化, 从而通过调控GM-CSF的信号转导过程, 促进CFU-GM的增殖。

关键词: 人参总皂甙; 粒单系祖细胞; 粒-巨噬细胞集落刺激因子; 粒-巨噬细胞集落刺激因子受体α; 信号转导

中图分类号: R331.1

The modulating mechanism for expression of human GM-CSF and GM-CSFRαby total saponins of Panax Ginseng

WANG Sha-Li1, CHEN Di1, WANG Ya-Ping2, LIU Yong-Gang2, JIANG Rong2

1Department of Physiology, College of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016

Abstract:The purpose of the present study was to investigate the biological mechanism for modulating granulocytopoiesis by Panax Ginseng. The techniques of culture of hematopoietic progenitor cells and hematopoietic stromal cells in vitro, biological assay of hematopoietic growth factor (HGF), immunocytochemistry, nucleic acid in situ hybridization, immunoprecipitation and Western blot were used to explore the effect of total saponins of Panax Ginseng (TSPG) on expression of human granulocyte-macrophage colony stimulating factor (GM-CSF) and granulocyte-macrophage colony stimulating factor receptor α (GM-CSFRα). The results indicated that (1) bone marrow stromal cell (BMSC),thymocyte (TC), splenocyte (SC), endothelial cells (EC), and monocyte (MO) conditioned media prepared with TSPG (50 μg/ml) could significantly enhance the proliferation of CFU-GM; (2) the expressions of GM-CSF in protein and mRNA level in BMSC, TC, SC, EC and MO induced by TSPG (50 μg/ml) were much higher than that of the control; (3) the expression of GM-CSFRα protein in hematopoietic cells induced by TSPG (50 μg/ml) was stronger than that of the control; (4) TSPG (50 μg/ml) could stimulate the transient tyrosine phosphorylation of GM-CSFR and Shc protein. We speculate that TSPG may directly and /or indirectly promote stromal cells in hematopoietic inductive microenvironment and lymphocytes to produce GM-CSF and other cytokine, induce bone marrow hematopoietic cells to express GM-CSF receptors (GM-CSFRα), regulate the GM-CSFR-mediated signals transductive pathway, and then proceed to proliferation of human CFU-GM.

Key words: total saponins of Panax Ginseng; CFU-GM; GM-CSF; GM-CSFR α; signal transduction