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目的探讨同基因骨髓移植(BMT)小鼠骨髓基质细胞内皮抑素(endostatin,ES)的表达及川芎嗪对其影响。方法取健康BALBc小鼠,随机分为3组正常组(不做处理),骨髓移植对照组(BMT组)和骨髓移植川芎嗪治疗组(川芎嗪组)。BMT组和川芎嗪组分别胃饲生理盐水02ml只和川芎嗪注射液每次2mg只,2次d。在BMT后第7、14、21、28天处死小鼠,采用免疫组化、Westernblot方法检测骨髓基质细胞中ES蛋白表达水平,RTPCR检测骨髓基质细胞中ESmRNA表达水平。结果BMT后第7、14、21、28天川芎嗪组和BMT组ESmRNA和蛋白表达水平均显著低于正常组(P<005或P<001),随着时间延长其表达逐渐增高。BMT后第14、21、28天川芎嗪组ES表达水平始终低于BMT组(P<005);但骨髓有核细胞计数高于BMT组(P<005或P<001)。结论川芎嗪能反馈调节BMT小鼠骨髓基质细胞新血管生成抑制因子的表达,促进骨髓微环境的修复,加速移植后骨髓的造血重建。
Objective To investigate the expression of endostatin (ES) in bone marrow stromal cells from syngeneic bone marrow transplantation (BMT) and the effects of ligustrazine on it. Methods Healthy BALBc mice were randomly divided into three groups: normal group (no treatment), bone marrow transplantation control group (BMT group) and bone marrow transplantation ligustrazine treatment group (Tetramethylpyrazine group). The rats in the BMT group and the Ligustrazine group were given 2 mg of normal saline and 2 mg of Ligustrazine each time for two days. The mice were killed on the 7th, 14th, 21st, and 28th days after BMT. The expression of ES protein in bone marrow stromal cells was detected by immunohistochemistry and Western blot. The expression of ESmRNA in bone marrow stromal cells was detected by RT-PCR. Results The mRNA and protein expression levels of ES mRNA and protein in the tetramethylpyrazine group and BMT group were significantly lower on the 7th, 14th, 21st, and 28th days after BMT (P<005 or P<001), and their expression gradually increased with time. The expression level of ES in the tetramethylpyrazine group was always lower than that in the BMT group on the 14th, 21st, and 28th days after BMT (P<005), but the nucleated cell count in bone marrow was higher than that in the BMT group (P<005 or P<001). Conclusion Tetramethylpyrazine can feedback regulate the expression of neovascularization inhibitors in bone marrow stromal cells of BMT mice, promote the repair of bone marrow microenvironment and accelerate the hematopoietic reconstitution of bone marrow after transplantation.