Sun Yanmei, Gao Fan, Xu Da, Lu Lei, Chen Qianggen, Yang Zheqing, Wang Xuenan, Pan Xiaoyan
Center for Reproductive Medicine, Jilin Medical University, Jilin 132013, China.
Department of Neonatology, Jilin Central General Hospital, Jilin 132011, China.
Evid Based Complement Alternat Med. 2021 Sep 27;2021:8035997. doi: 10.1155/2021/8035997. eCollection 2021.
Many ingredients in Wenshen Shengjing Decoction (WSSJD) can cause epigenetic changes in the development of different types of cells. It is not yet known whether they can cause epigenetic changes in sperms or early embryos. Here, we investigated the role of WSSJD in epigenetic modifications of sperms or early embryos and early embryo development. A mouse model with spermatogenesis disorders was established with cyclophosphamide (CPA). WSSJD was administrated for 30 days. The male model mice after the treatment were mated with the female mice treated with superovulation. The embryo development rate of each stage was calculated. Immunofluorescence staining was used to detect the expression of H3K27me3 in sperm, pronuclear embryos, and 2-cell embryos. Western blotting was used to detect the expression of histone demethylase KDM6A and methyltransferase EZH2 in 2-cell embryos with developmental arrest. The expressions of zygotic genome activation genes (, , , , and ) in 2-cell embryos with developmental arrest were analyzed with qRT-PCR. Comparing with the control group, CPA destroyed the development of seminiferous epithelium, significantly increased the expression level of H3K27me3 in sperm, reduced the expression ratio of H3K27me3 in female and male pronuclei, delayed the development of 2-cell embryos, and increased the developmental arrest rate and degeneration rate of 2-cell embryos. Moreover, the expressions of EZH2 and H3K27me3 were significantly increased in the 2-cell embryos with developmental arrest, and the expression of zygotic genome activation genes (, , , , and ) was significantly decreased. Compared with the CPA group, WSSJD promoted the development of seminiferous epithelium, maintained a low level of H3K27me3 modification in sperm and male pronucleus, significantly increased the development rate of 2-cell embryos and 3-4 cell embryos, and reduced the developmental arrest rate and degeneration rate of 2-cell embryos. WSSJD may promote early embryonic development by maintaining a low level of H3K27me3 modification in sperm and male pronucleus and regulating the zygotic genome activation in mice with spermatogenesis disorders induced by CPA.
温肾生精汤（WSSJD）中的多种成分可在不同类型细胞的发育过程中引起表观遗传变化。目前尚不清楚它们是否会导致精子或早期胚胎发生表观遗传变化。在此，我们研究了温肾生精汤在精子或早期胚胎的表观遗传修饰以及早期胚胎发育中的作用。用环磷酰胺（CPA）建立了精子发生障碍的小鼠模型。给予温肾生精汤30天。将处理后的雄性模型小鼠与经超排卵处理的雌性小鼠交配。计算各阶段的胚胎发育率。采用免疫荧光染色检测精子、原核胚胎和2细胞胚胎中H3K27me3的表达。采用蛋白质免疫印迹法检测发育阻滞的2细胞胚胎中组蛋白去甲基化酶KDM6A和甲基转移酶EZH2的表达。用qRT-PCR分析发育阻滞的2细胞胚胎中合子基因组激活基因（、、、、和）的表达。与对照组相比，CPA破坏了生精上皮的发育，显著提高了精子中H3K27me3的表达水平，降低了雌雄原核中H3K27me3的表达比例，延迟了2细胞胚胎的发育，并增加了2细胞胚胎的发育阻滞率和退化率。此外，发育阻滞的2细胞胚胎中EZH2和H3K27me3的表达显著增加，合子基因组激活基因（、、、、和）的表达显著降低。与CPA组相比，温肾生精汤促进了生精上皮的发育，维持了精子和雄性原核中低水平的H3K27me3修饰，显著提高了2细胞胚胎和3-4细胞胚胎的发育率，降低了2细胞胚胎的发育阻滞率和退化率。温肾生精汤可能通过维持精子和雄性原核中低水平的H3K27me3修饰以及调节CPA诱导的精子发生障碍小鼠的合子基因组激活来促进早期胚胎发育。
