地榆总皂苷对巨核祖细胞增殖分化及相关受体表达的影响

Effect of total saponins from Sanguisorba officinalis on megakaryocyte progenitor cells proliferation,differentiation and relative receptor expression

目的:观察地榆总皂苷(DYS)对Baf3细胞和32D细胞的促增殖和分化作用,以及对IL-3受体和干细胞因子受体c-kit表达水平的影响。方法:培养依赖IL-3生长的Baf3细胞和32D细胞,在加或不加IL-3条件下,用质量浓度为5,10,20,30,40 mg·L-1的DYS分别处理细胞24,48,72,96 h后,采用CCK8方法检测细胞增殖;并用Giemsa染色检测细胞分化;另以RT-PCR方法检测DYS对Baf3细胞IL-3受体及对32D细胞c-kit表达水平的影响。结果:DYS单独处理细胞48 h,明显促进Baf3细胞和32D细胞增殖;与对照细胞比较,DYS处理32D细胞呈现成簇生长并形成许多大的细胞团;在加入IL-3的条件下,DYS也能明显增强IL-3的促细胞增殖作用。此外,DYS高浓度单独处理32D细胞可明显诱导多倍体巨核细胞数增加,促进巨核细胞分化。在mRNA水平,DYS单独处理细胞也能明显上调IL-3受体和c-kit的表达。结论:地榆总皂苷单独作用能促使2种巨核祖细胞的增殖和分化,其作用与上调IL-3受体和c-kit表达水平有关。

Objective： To investigate the effects of total saponins from Sanguisorba officinalis （DYS） on hematopoietic cell proliferation, differentiation and the expression level of IL-3R and c-kit. Method： Baf3 and 32D ceils were cultured with or without IL-3, then the cells were exposed to DYS in different concentrations of 5, 10, 20,30 and 40 mg· L^-1 for 24,48,72 and 96 hours sepa- rately. After that, the cell proliferation and differentiation capacity were determinated by the methods of CCK8 and Giemsa staining sep- arately. The effects of DYS on the expression level of IL-3 receptor in Bar3 cells and the expression level of c-kit in 32D cells were determinated using RT-PCR. Result： DYS promotes alone proliferation of Baf3 cells and 32D cells after 48 h. In contrast to control cells, 32D cells containing DYS without IL-3 form many large clusters. DYS also increases the proliferation when cultured with IL-3. High concentration of DYS induce alone the differentiation of 32D cells and increase alone the number of the polyploidy megakaryocyte. Mo- reover, DYS increases alone the expression level of IL-3R in Bar3 cells and the expression level of c-kit in 32D cells separately. Con- clusion： Our data shows DYS can promote alone proliferation and differentiation of megakaryocyte progenitor cells. The proliferative and differentiative effect of DYS on megakaryocyte progenitor cells is correlated to the up-regulation of IL-3 receptor and c-kit expression.