Department of Bioscience Technology and Center for Nanotechnology, Chung Yuan Christian University, Chung Li 32023, Taiwan.
Reprod Toxicol. 2009 Dec;28(4):530-5. doi: 10.1016/j.reprotox.2009.07.004. Epub 2009 Jul 29.
In this report, we examine the cytotoxic effects of puerarin, an isoflavone compound, on the blastocyst stage of mouse embryos and subsequent embryonic attachment and outgrowth in vitro and in vivo implantation by embryo transfer. Mouse blastocysts were incubated in medium with or without puerarin (2.5, 5 or 10 microM) for 24h. Cell proliferation and growth was investigated by dual differential staining, apoptosis was analyzed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and implantation and post-implantation development of embryos were measured by in vitro development analysis and in vivo embryo transfer, respectively. We found that blastocysts treated with 5 or 10 microM puerarin showed significant increases in apoptosis and significant decreases in total cell number. Interestingly, there was no significant difference of implantation success rate between puerarin-pretreated blastocysts and controls, but in vitro treatment with 5 or 10 microM puerarin was associated with increased resorption of post-implantation embryos and decreased fetal weight. Our results collectively indicate that in vitro exposure to puerarin induced apoptosis and after transfer to host mice retards early post-implantation development. The extent to which puerarin may have teratogenic potential in early human development is not yet known.
在本报告中，我们研究了葛根素（一种异黄酮化合物）对小鼠胚胎囊胚期的细胞毒性作用，以及随后通过胚胎转移进行体外和体内胚胎附着和生长的情况。将小鼠囊胚在含有或不含有葛根素（2.5、5 或 10μM）的培养基中孵育 24 小时。通过双重差异染色研究细胞增殖和生长，通过末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）分析细胞凋亡，通过体外发育分析和体内胚胎转移分别测量胚胎的植入和植入后发育。我们发现，用 5 或 10μM 葛根素处理的囊胚显示出明显的凋亡增加和总细胞数明显减少。有趣的是，预处理囊胚与对照组之间的植入成功率没有显著差异，但在体外用 5 或 10μM 葛根素处理与植入后胚胎吸收增加和胎儿体重下降有关。我们的研究结果表明，体外暴露于葛根素诱导细胞凋亡，并在转移到宿主小鼠后延迟早期植入后发育。葛根素在人类早期发育中是否具有致畸潜力尚不清楚。
