失血性贫血小鼠恢复过程中骨髓基质金属蛋白酶变化

目的:观察失血性贫血小鼠恢复过程中骨髓基质金属蛋白酶-2和-9(MMP-2,MMP-9)的变化,并探讨其在造血调控中的作用.方法:采用全自动血细胞分析仪、免疫组化和酶谱电泳法分别检测失血性贫血小鼠恢复过程中外周血RBC和Hb数量、骨髓细胞MMP-2和MMP-9表达以及骨髓造血微环境MMP-2和MMP-9的活性变化.结果:(1)与正常对照组相比外周血RBC和Hb数量失血后第1d急剧下降.随着恢复时间的延长,二者数目逐渐上升,到第9d已接近正常对照组水平.(2)在正常对照组检测到了较弱的MMP-2和MMP-9的表达.与正常对照组相比,失血后1d组、3d、5d和7d组小鼠骨髓细胞中MMP-2和MMP-9表达明显增加,MMP-2和MMP-9表达分别于失血后5d和3d达到峰值.与1d组相比,3d组小鼠骨髓细胞中MMP-2和MMP-9表达以及5d组小鼠骨髓细胞中MMP-2表达明显增加.(3)正常对照组中检测到proMMP-2,proMMP-9和MMP-9 3条酶带,MMP-9的活性最强.失血后1d,proMMP-2,proMMP-9和MMP-9的活性均急剧升高.随着恢复时间的延长,3者的活性逐渐降低,到第9d时恢复到接近正常对照组水平.结论:失血性贫血小鼠可能通过骨髓细胞中MMPs表达增加以及骨髓造血微环境中MMPs活性升高来促进骨髓造血功能增强,使外周血RBC和Hb的数量恢复到正常水平.

Changes of Bone Marrow Matrix Metalloproteinases During the Recovery Process of Blood Loss Anemia Mice

Objective: To examine the changes of the expression and activities of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9) in bone marrow(BM)of blood loss anemia mice,and to explore its mechanism for hematopoietic regulation.Methods: The changes of peripheral blood RBC and Hb,the expressions and activities of MMP-2 and MMP-9 in bone marrow(BM)of blood loss anemia mice were detected by automatic blood cell analysator,immunohistochemistry method and gelatin zymography respectively.Results: It was found that(1) the number of RBC and Hb content were rapid decreased in day 1(1 d)group after blood loss.Compared with control group,there were significant difference.Along with the prolongation of recovery time,both RBC and Hb number increased generally and approached the levels of control group at 9 d.(2) the expression of MMP-2 and MMP-9 were weaker in control group.Compared with control group,the expression of MMP-2 and MMP-9 was obviously increased in 1 d,3 d,5 d and 7 d groups,and reached peak value at 5d and 3d respectively after blood loss.Compared with 1d group,the expression of MMP-2 and MMP-9 was obviously increased in 3d group,and the expression of MMP-2 was obviously increased in 5 d group.(3) gelatin zymography revealed that proMMP-2,proMMP-9 and MMP-9 were detected in control group,and the activity of MMP-9 was stronger among them.At 1d after blood loss,the activities of proMMP-2,proMMP-9 and MMP-9 were rapid elevated,and their activities decreased generally along with the prolong of recovery time,and approached the levels of control group at 9d.Conclusion: These results suggest that the peripheral blood RBC and Hb number returned to normal levels were caused by increasing the expressions and activities of MMP-2 and MMP-9 in BM of blood loss anemia mice to enhance hematopoietic function of BM.