摘要
Objective:To explore the effect and mechanism of Xuebijing injection (XBJ) on the hematopoietic homeostasis of bone marrow (BM) in septic mice. Methods:BM cells stimulated with XBJ were analyzed by inverted optical microscope and qPCR, to evaluate the effect of XBJ on differentiation function of BM cells in vitro. Lipopolysaccharide (LPS) 055:B5 at the dose of 20mg/kg was used to establish the sepsis model. To determine the hematopoietic stem cells homeostasis affected by LPS in mice, BM cells were isolated and analyzed by flow cytometry based on the immunophenotypic surface markers. 20ml/kg of XBJ was administered by tail vein once/day after peritoneal injection of LPS. All animals were sacrificed on the fifth day. The frequency of hematopoietic stem cells (HSC) and immune cells in mice were quantified by flow cytometry. The gene expression of transcription factors were detected by qPCR. Results:XBJ promoted myeloid differentiation of BM cells in vitro, which may be related to the mRNA expression of the transcription factors. The results in vivo showed the percentage of BM Lin-SCA-1+c-KIT+(LSK) cells, and Long-term HSCs (LT-HSC) were significantly increased in LPS group. But the percentage of multipotential progenitors (MPPs), granulocyte-monocyte progenitor (GMP) and megakaryocytic-erythroid precursor (MEP) were significantly decreased in LPS group. Whereas, XBJ improved the immune function in sepsis mice by suppression the LSK expansion in vivo. The result of qPCR showed that LSD1 and PU.1 mRNA expression in XBJ group were significantly higher than LPS group and control group respectively. Conclusion:Xuebijing injection can improve immune function in sepsis mice by regulating hematopoietic homeostasis. Its mechanism may be related to the up-regulation of LSD1 and PU.1 gene expressions.
目的:研究血必净注射液对脓毒症小鼠骨髓造血稳态的影响及其机制。方法:血必净注射液刺激离体小鼠骨髓细胞,观察其对骨髓细胞增殖分化功能的影响。采用20mg/kg 剂量的LPS 构建脓毒症小鼠模型,流式细胞术检测LPS 对小鼠骨髓造血干细胞及祖细胞含量的影响。腹腔注射LPS 后,采用尾静脉注射的方法每天给予20ml/kg 的血必净注射液治疗,第5 天处死动物,从腿骨提取骨髓细胞,利用流式细胞术检测血必净注射液对脓毒症小鼠骨髓造血干细胞及免疫细胞的影响,通过荧光定量PCR 检测血必净注射液对小鼠骨髓细胞转录因子的影响。结果:体外实验结果显示,血必净注射液可通过上调CD11B 基因的表达,下调GFI1B、GATA2 及FOS 基因的表达,促进小鼠骨髓细胞的分化。动物实验结果发现,LPS 诱导了小鼠骨髓造血干细胞LSK 的病理性增殖,导致多能祖细胞MPP、粒系-巨噬系祖细胞GMP 及巨核系-红系祖细胞MEP 的含量显著降低,从而改变了小鼠骨髓的造血稳态。而血必净注射液可通过抑制LSK 的病理性增殖,抑制髓源性抑制细胞MDSC 的含量,改善脓毒症小鼠的免疫功能。其机制可能与其上调LSD1 及PU.1 基因的表达有关。结论:血必净注射液可通过调节脓毒症小鼠骨髓细胞的造血稳态改善免疫功能,其机制可能与其上调LSD1 及PU.1 基因的表达有关。
