Effects of methotrexate on the developments of heart and vessel in zebrafish
Effects of methotrexate on the developments of heart and vessel in zebrafish
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DOI：
10.1093/abbs/gmn010
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发表时间：
2009-01-01
影响因子：
3.7
通讯作者：
Song, Houyan
中科院分区：
文献类型：
--
作者：
Sun, Shuna;Gui, Yonghao;Song, Houyan
叶酸拮抗剂甲氨蝶呤(MTX)能抑制二氢叶酸还原酶(DHFR)，DHFR在四氢叶酸的合成和胚胎发育中起重要作用。在本研究中，我们发现斑马鱼胚胎在受精后6-10小时暴露于1.5 mM的MTX后，无法形成正常的心血管系统。在MTX处理的胚胎中，心室和心房的形态发育被破坏，心脏扭转异常，心率和心室短轴缩短率降低，血管发育受阻。我们还发现，无论是微量注射DHFR-GFP mRNA还是用五水合叶酸钙盐(CF)处理，都能促进MTX处理的胚胎心脏和血管的发育，这证明MTX是通过抑制DHFR而导致畸形的。在MTX处理的胚胎中，hand2、mef2a、mef2c和flk-1基因的转录水平降低。与MTX处理组相比，MTX 1DHFR-GFP mRNA注射组和MTX 1CF组Hand2、mef2a、MEF2C和Flk-1的转录水平均升高。我们的结果表明，MTX处理的胚胎心脏和血管的发育受阻与Hand2、mef2a、mef2c和flk-1的转录水平降低有关。
Methotrexate (MTX), an antagonist of folic acid, can inhibit dihydrofolate reductase (DHFR) which is of great importance in the synthesis of tetrahydrofolic acid and embryonic development. In this study, we found that after being exposed to 1.5 mM MTX at 6-10 hours post-fertilization, zebrafish embryos fail to form normal cardiovascular system. In MTX-treated embryos, the morphological development of ventricle and atrium was disrupted, the cardiac twist was abnormal, the heart rate and ventricular shortening fraction were reduced, and the vascular development was disrupted. We also found that either microinjection with dhfr-gfp mRNA or treatment with folinic acid calcium salt pentahydrate (CF) could cause improved development in the heart and vessels in MTX-treated embryos, which proved that MTX induced the malformations by inhibiting DHFR. The transcript levels of genes such as hand2, mef2a, mef2c, and flk-1 were reduced in MTX-treated embryos. Compared with the MTX-treated group, the transcript levels of hand2, mef2a, mef2c, and flk-1 were increased in the MTX 1 dhfr-gfp mRNA-injected group and in the MTX 1 CF group. Our results indicated that the disrupted development of the heart and vessels in MTX-treated embryos is related to the reduced transcript levels of hand2, mef2a, mef2c, and flk-1.