三七总皂苷对斑马鱼胚胎初级和次级造血的抑制作用

Panax notoginsenosides inhibit primitive and definitive hematopoiesis during the zebrafish embryonic development

目的探讨三七总皂苷(Panax notoginsenosides,PNSs)对斑马鱼胚胎造血的作用,为三七的药理学应用提供实验依据。方法通过乙醇提取得到PNSs,用50、100μg/mL的PNSs从75%外包时期开始处理斑马鱼胚胎。收集发育至不同时期的胚胎,检测药物处理后,斑马鱼初级造血和次级造血的分子标记的变化。结果 PNSs处理后,初级造血的分子标记gata1、hbbe3明显下降,生成的红细胞显著减少;PNSs处理还可以抑制造血干细胞(hematopoietic stem cell, HSC)发育。次级造血的分子标记runx1,cmyb在PNSs处理下表达下调,由HSC分化生成的T淋巴细胞分子标记rag1表达也显著降低。有意思的是,PNSs对斑马鱼初级和次级造血的抑制作用均呈剂量依赖效应。结论孕期可能需要慎重使用三七总皂苷药物。

Objective To investigate the effects of Panax notoginsenosides (PNSs) on primitive and definitive hematopoiesis of zebrafish embryos, and provide new insights into pharmacological application of PNSs. Methods PNSs were obtained by ethanol extraction from panax notoginseng powder. Zebrafish embryos were treated with 50 or 100 μg/mL PNSs from the 75% epiboly stage until harvest to detect hematopoietic markers by in situ hybridization. Results Compared with the control larvae, the embryos treated with PNSs exhibited obviously reduced expression of primitive hematopoietic markers such as gata1 and hbbe3. Consistently, the formation of erythrocytes was also impaired in the PNSs-treated embryos at later stages. Furthermore, PNSs treatments disrupted the definitive hematopoiesis as indicated by decreased expression of hematopoietic stem cell markers runx1 and cmyb and the T lymphocyte marker rag1. Interestingly, the inhibitory effects of PNSs on the primitive and definitive hematopoiesis were concentration-dependent. Conclusions Our results indicate that PNSs impair primitive and definitive hematopoiesis during zebrafish embryonic development, implying that PNSs should be used with caution during pregnancy.