摘要
目的:探讨川芎嗪对放射损伤小鼠骨髓基质细胞(BMSCs)中血管内皮生长因子(VEGF)表达的影响及其意义。方法:采用蛋白免疫印迹(Westernblot)法、免疫组化法及流式细胞术,对放射损伤小鼠BMSCs中VEGF蛋白表达水平、骨髓组织中VEGF受体胎肝激酶1(flk1)表达变化、BMSCs凋亡率及细胞周期改变进行分析,同时观察川芎嗪的影响。结果:放射损伤后小鼠BMSCs中VEGF和骨髓组织中flk1表达均明显低于正常水平,随时间推移而逐渐升高,但照射后第14d仍未恢复正常;而川芎嗪组在第14d时已接近正常水平。放射损伤后BMSCs停滞于G0/G1期,S期合成减少,凋亡率明显增加;随时间推移G0/G1期细胞比例呈由高到低变化,凋亡率也逐渐降低,但第14d时仍未恢复正常;用川芎嗪治疗后,BMSCs的S期合成活跃,凋亡率明显下降,第14d时恢复更明显,接近正常水平。骨髓切片苏木素伊红(HE)染色也证实川芎嗪组小鼠造血恢复明显较对照组快。结论:川芎嗪促进BMSCs中VEGF的表达,通过VEGF/flk1途径改善放射损伤小鼠骨髓微环境,是其促进造血功能恢复的机制之一。
Objective： To evaluate the effect of tetramethylpyrazine on the expression of vascular endothelial growth factor （VEGF） in bone marrow stromal cells （BMSCs） in mice with radiation injury and its significance. Methods： The protein expression of VEGF in BMSCs was assayed by Western blot, each paraffin-embedded section was stained with anti -fetal liver kinase - 1 （flk - 1） ; and the cell cycle and apoptosis rate of BMSCs were tested by flow cytometer （FCM）. The effect of tetramethylpyrazine on the hematopoiesis was evaluated at the same time. Results ： The expression levels of VEGF in BMSCs and flk - 1 in bone marrow tissue decreased significantly after radiation and increased slowly afterwards. The value in tetramethylpyrazine group almost reached normal level, while it remained lower in control group on 14 th day. After ^60Co ray irradiation, the BMSCs were arrested in G0/G1 phase, apoptosis rate increased significantly. These values recovered slowly with the time and remained higher than those in normal group on 14 th day. The recovery of these values in tetramethylpyrazine -treated group was sooner than those in radiation group, and they almost reached the normal levels on 14 th day. The condition in the hematoxylin and eosin （HE） stained bone marrow section also suggested that the recovery of hematopoiesis in tetramethylpyrazine - treated group was sooner than that in the control group. Conclusion： The tetramethylpyrazine can enhance the expression of VEGF in BMSCs, and improve bone marrow microenvironment through VEGF/flk - 1 pathways. This may be one of the mechanisms of tetramethylpyrazine to promote recovery of hematopoiesis after radiation.
出处
《中国中西医结合急救杂志》
CAS
2005年第4期195-198,共4页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
国家自然科学基金资助项目(39870926)
关键词
川芎嗪
放射损伤
骨髓微环境
血管内皮生长因子
胎肝激酶-1
tetramethylpyrazine
radiation injury
bone marrow microenvironment
vascular endothelial growth factor
fetal liver kinase - 1
