川芎嗪改善免疫诱导再生障碍性贫血小鼠骨髓微环境作用的研究

Study on the improvement of bone marrow microenvironment by ligustrazine in immuneinduced aplastic anemia mice

目的：探讨川芎嗪对再生障碍性贫血（再障）骨髓微环境的作用及其机制。方法：建立免疫介导的再障小鼠模型，胃饲川芎嗪注射液每次４ｍｇ，１天２次，第１０天用氧分压传感针测定活体尺骨中段骨髓氧分压（ＰｂＯ２），再观察其骨髓切片组织学，体外成纤维细胞集落形成单位（ＣＦＵ－Ｆ），培养基质细胞层的粘附能力。结果：川芎嗪组ＰｂＯ２为１０．３２±１．２７ｋＰａ，显著高于再障组（４．３２±２．８６ｋＰａ），Ｐ＜０．００１。再障组骨髓微血管扩张、断裂、瘀血，造血组织容量百分率为２４．９％±９．６％，ＣＦＵ－Ｆ为１２．５±７．３／２×１０６骨髓有核细胞（ＢＭＮＣ）。川芎嗪组骨髓微血管较清楚、完整，无断裂及瘀血，造血组织容量百分率为５２．８％±１５．６％，ＣＦＵ－Ｆ为３１．５±１０．６／２×１０６ＢＭＮＣ。川芎嗪组体外培养基质细胞粘附正常小鼠骨髓有核细胞的能力为７２．７％±７．８％，与正常组（７３．４％±３．４％）无差异，明显高于再障组（５６．２％±９．８％），Ｐ＜０．０１。结论：川芎嗪能促进再障小鼠骨髓微血管的修复，增加对骨髓的供氧，促进骨髓基质细胞生长及其粘附功能。川芎嗪通过改善骨髓微环境而使骨髓造血细胞增生。

Objective: To explore the effects of ligustrazine on bone marrow microenvironment and its mechanism in aplastic anemia (AA). Methods:Each immuneinduced AA mouse was gastric fed by 4mg ligustrazine twice a day. On the 10th day, the ulnar bone marrow partial pressure of oxygen (PbO2) was determined in vivo by a PO2 sensory needle. Then the histological features, fibroblastic colony forming unit (CFUF) yields and the adhesive function of stromal cells of the bone marrow were assayed in vitro. Results: The PbO2 in ligustrazinetreated group was 10.32±1.27kPa, while in AA group was 4.32±2.86kPa(P<0.001). In AA group, the microvessels were expanded, broken and being stasis. The percentage of hematopoietic tissue volume was 24.9%±9.6% and the CFUF yields was 12.5±7.3/2×106 BMNC. The microvessels in ligustrazine group were more clear and intact, not being broken and had no stasis. The percentage of hematopoietic tissue volume was 52.8%±15.6% and the CFUF yields was 31.5±10.6/2×106 BMNC. In ligustrazine group, the adhensive function of stromal cell layer cultured with bone marrow nucleated cells from normal mice was 72.7%±7.8%,which was not different from that in normal group(73.4%±3.4%),but much higher than that in AA group(56.2%±9.8%,P<0.01). Conclusion:Ligustrazine can promote the rehabilitationof bone marrow microvessels in AA mice, increasing the oxygen supply for bone marrow microenvironment, promoting the growth of stromal cells and strengthening their adhesive function. Ligustrazine enbances the bone marrow hematopoietic cells proliferation by improving their microenvironment.