Center for Reproductive Medicine, Jilin Medical University, Jilin 132013, China.
Department of Urology Surgery, The Affiliated 465 Hospital of Jilin Medical University, Jilin 132013, China.
Biomed Res Int. 2022 Feb 21;2022:9775473. doi: 10.1155/2022/9775473. eCollection 2022.
New Wenshen Shengjing Decoction (NWSSJD), a traditional Chinese compound medicine, has significant effect on spermatogenesis disorder and can significantly improve sperm quality. Many components in NWSSJD can induce epigenetic modifications of different types of cells. It is not yet known whether they can cause epigenetic modifications in sperm or early embryos.
This study investigated the effect of NWSSJD on mouse early embryonic development and its regulation of H3K4me3 in mouse sperm and early embryos.
Spermatogenesis disorder was induced in male mice with CPA (cyclophosphamide). NWSSJD was administrated for 30 days. Then, the male mice were mated with the female mice with superovulation, and the embryo degeneration rate of each stage was calculated. Immunofluorescence staining was used to detect the expression of H3K4me3 in sperm and embryos at various stages. Western blotting was performed to detect methyltransferase SETD1B expression. The expressions of development-related genes (, , and ) and apoptosis-related genes ( and ) were measured with qRT-PCR.
Compared with the CPA group, NWSSJD significantly reduced the H3K4me3 level in sperms, significantly increased the number of normal early embryos (2-cell embryos, 3-4-cell embryos, 8-16-cell embryos, and blastocysts) per mouse, and reduced the degeneration rate of the embryos. The expression levels of H3K4me3 and methyltransferase SETD1B in early embryos were significantly elevated by NWSSJD. Additionally, NWSSJD significantly promoted expression, while reducing expression, thus inhibiting embryonic cell apoptosis. Moreover, the expressions of development-related genes and were significantly increased by NWSSJD, but expression had no significant difference.
NWSSJD may promote early embryonic development possibly by maintaining low H3K4me3 levels in sperms and normal H3K4me3 modification in early embryos and by inhibiting embryonic cell apoptosis.
新温肾生精方（NWSSJD）是一种中药复方，对精子发生障碍有显著疗效，能显著改善精子质量。NWSSJD 中的许多成分可以诱导不同类型细胞的表观遗传修饰。目前尚不清楚它们是否会引起精子或早期胚胎的表观遗传修饰。
本研究探讨了 NWSSJD 对小鼠早期胚胎发育的影响及其对小鼠精子和早期胚胎中 H3K4me3 的调控作用。
用环磷酰胺（CPA）诱导雄性小鼠精子发生障碍。给予 NWSSJD 治疗 30 天。然后，雄性小鼠与超排卵的雌性小鼠交配，计算各阶段胚胎退化率。免疫荧光染色检测精子和各阶段胚胎中 H3K4me3 的表达。Western blot 检测甲基转移酶 SETD1B 的表达。用 qRT-PCR 检测发育相关基因（、和）和凋亡相关基因（和）的表达。
与 CPA 组相比，NWSSJD 显著降低精子中的 H3K4me3 水平，显著增加每只小鼠正常早期胚胎（2 细胞胚胎、3-4 细胞胚胎、8-16 细胞胚胎和囊胚）的数量，降低胚胎退化率。NWSSJD 显著上调早期胚胎中 H3K4me3 和甲基转移酶 SETD1B 的表达。此外，NWSSJD 显著促进 表达，降低 表达，从而抑制胚胎细胞凋亡。而且，NWSSJD 显著增加发育相关基因 和 的表达，但 的表达无显著差异。
NWSSJD 可能通过维持精子中低水平的 H3K4me3 和早期胚胎中正常的 H3K4me3 修饰，并抑制胚胎细胞凋亡，从而促进早期胚胎发育。
