急性白血病患者骨髓血管新生的研究及临床意义

The clinical significance of angiogenesis in the bone marrow of acute leukemia patients

目的 研究急性白血病 (AL)患者骨髓血管新生、骨髓液血管内皮生长因子 (VEGF)水平并探讨其临床意义。方法 采用免疫组化方法 ,以兔抗人Ⅷ相关抗原多克隆抗体标记骨髓内皮细胞 ,光学显微镜下计数骨髓全切片微血管数 ;用ELISA方法检测骨髓液VEGF水平。结果 骨髓微血管数在初发未治组 (2 2 82 /× 4 0 0视野 )明显高于正常对照组 (7 17/× 4 0 0视野 )及骨髓缓解组(8 5 7/× 4 0 0视野 ) (P值均 <0 0 0 1) ,而骨髓缓解组仍高于正常对照组 (P <0 0 5 ) ,初发未治组与复发难治组 (2 1 83/× 4 0 0视野 )之间的差异无显著性 (P >0 0 5 )。初发未治组中 ,骨髓微血管数在 2 3例急性淋巴细胞白血病 (ALL)患者 (2 3 0 9/× 4 0 0视野 )与 4 3例急性非淋巴细胞白血病 (ANLL)患者(2 2 37/× 4 0 0视野 )、FAB亚型M1～ 3 (2 2 91/× 4 0 0视野 )与M4～ 5患者 (2 1 4 6 /× 4 0 0视野 )之间的差异均无显著性 (P值均 >0 0 5 ) ,而骨髓微血管数与骨髓原始细胞百分数呈正相关性 (r =0 311,P <0 0 1)。骨髓液VEGF水平 ,在初发未治组 (188 88ng/L)显著高于骨髓缓解组 (78 74ng/L)和正常对照组 (79 5 2ng/L) (P值均 <0 0 1) ,而后两者之间的差异无显著性 (P >0 0 5 ) ;在ANLL组(2 70 12ng/L)显著高于ALL组

Objective To study the angiogenesis in bone marrow and the level of vascular endothelial growth factor (VEGF) in bone marrow fluid from acute leukemia (AL) patients and to explore their clinical significance. Methods Microvessels in each bone marrow specimen section were counted by immunohistochemical identification of microvascular endothelial cells with anti-human von Willbrand factor under light microscopy in field 400 times. The VEGF level in bone marrow fluid was measured by ELISA. Results Microvessel counts in untreated AL patients (22.82/×400 field) were significantly higher than those in normal controls (7.17/×400 field) and in the bone marrow remission (BMR) AL patients (8.57/×400 field) ( P <0.001,both), while the controls in the latter were higher than those in controls ( P <0.05). There was no difference in microvessel counts between untreated AL patients and relapse/refractory AL patients (21.83/×400 field) ( P >0.05). In the untreated AL group, the microvessel counts showed difference neither between 23 ALL patients(23.09/×400 field) and 43 ANLL patients (22.37/×400 field) ( P >0.05), nor between patients with M_ 1-3 (22.91/×400 field ) and M_ 4-5 (21.46/×400 field) the two subtypes of FAB ( P >0.05). There was a positive correlation between the microvessel counts and the percentage of blast cells in the bone marrow ( r =0.311, P <0.01). The VEGF level in the bone marrow fluid in untreated AL patients (188.88 ng/L) was significantly higher than that in BMR AL patients (78.74 ng/L) and controls (79.52 ng/L) ( P <0.01), while the latter two groups had no difference between each other ( P >0.05). The VEGF level in the bone marrow fluid from ANLL group (270.12 ng/L) was enhanced significantly compared with that from ALL group (101.70 ng/L) ( P <0.01) and associated with the microvessel counts ( r =0.464, P <0.05). Conclusion Microvessel counts increased in the bone marrow in patients with different types of AL, suggesting that angiogenesis might play an important role in the pathology of AL. Testing the VEGF level in the bone marrow fluid from ANLL patients could reflect the degree of angiogenesis and disease condition.