环磷酰胺所致血虚证小鼠骨髓CD34^+细胞的变化

The changes of CD34^+ cells in bone marrow of blood deficiency mice induced by cyclophosphamide

目的 :研究环磷酰胺 (CTX)诱发的血虚证小鼠骨髓中干祖细胞的数量和骨髓细胞周期的动态变化。方法 :采用CTX大小两个剂量分别对两组小鼠进行腹腔注射制作小鼠血虚证模型。在不同时相点 ,双荧光标记测定CD34+ 细胞在骨髓有核细胞中的比例 ,骨髓细胞固定后PI染色测定细胞周期。结果 :①注射CTX后CD34+ 细胞在骨髓有核细胞中的比例下降后迅速上升 ,然后又下降。②注射CTX后骨髓细胞增殖受抑 ,然后动员骨髓细胞增殖 ,但药后 10d细胞增殖又受抑。与CD34+ 细胞在骨髓有核细胞中比例的变化规律有一致性。结论 :CTX可以动员骨髓造血干祖细胞进入细胞周期而增殖 ,增加了骨髓中造血干祖细胞的数量 ,但此过程加速了干祖细胞池的耗竭 ,从而对骨髓造成更大损伤 ,这可能是CTX诱发小鼠血虚证的机理之一。

Objective:To observe the changes of CD34 + cells and characterize the cell cycle status of bone marrow cells of blood deficiency mice induced by cyclophosphamide (CTX). Methods:Establishment of the blood deficiency model in mice by injected i.p with CTX. Flow cytometric enumeration of CD34 +hematopoietic stem and progenitor cells by double fluorescent labeling, and cell cycle detection by PI labeling at the indicated detection time. Results:①Compared with normal group, the rate of CD34 + cells in bone marrow nuclear cells decreased at first, then increased for some days, and then decreased. ②Bone marrow cell cycle perturbed after CTX injected, a large fraction of bone marrow cells are relatively more quiescent at first, then are in an actived state and proliferated, then a significant increase in the number of quiescent bone marrow cells. These changes matched those changes of CD34 + cells. Conclusion: CTX mobilize more hematopietic stem and progenitor cells into S/G2/M cycle, and the number of stem and progenitor cells increased, this result in the exhaustion of pool of stem and progenitor cells and a lot of damage to bone marrow. This may be one of causes of blood deficiency induced by CTX.