ICAR-National Institute of Animal Nutrition and Physiology, Adugodi, Hosur Road, Bengaluru 560 030, India.
ICAR-National Institute of Animal Nutrition and Physiology, Adugodi, Hosur Road, Bengaluru 560 030, India.
J Reprod Immunol. 2023 Sep;159:104133. doi: 10.1016/j.jri.2023.104133. Epub 2023 Aug 15.
The effect of interleukin-6 (IL-6) supplementation during the different phases of in vitro embryo culturing (IVC) on embryo development and embryonic gene expression was studied in ovine. IL-6 was added to IVC medium during the late phases (72-192 h; 5, 10, and 25 ng/ml IL-6) or entire period (0-192 h; 10 ng/ml IL-6) of IVC to determine its effect on embryo development. Further, the effect of IL-6 (10 ng/ml) supplementation at the 72 h of IVC on gene expressions associated with JAK/STAT signalling and pluripotency in 8-16 cell embryos (1 h post-supplementation) and compact morulae (48 h post-supplementation), and apoptosis and primitive endoderm (PrE) development in compact morulae was investigated. The supplementation of 10 ng/ml IL-6 during the late phases of IVC significantly (P < 0.05) increased blastocyst formation (35.2 ± 1.52%) compared to the control (21.1 ± 1.11%), and 5 ng/ml (25.9 ± 2.98%) or 25 ng/ml (16.5 ± 0.73%) IL-6 groups. Conversely, IL-6 (10 ng/ml) treatment throughout the IVC period significantly (P < 0.05) decreased the rate of cleavage (55.4 ± 1.57%) and blastocyst formation (14.5 ± 1.28%) compared to the control group (65.8 ± 1.35% and 21.5 ± 0.97%, respectively). In 8-16 cell embryos and compact morulae, the IL-6 treatment significantly (P < 0.05) affected the expression of genes associated with JAK/STAT signalling and pluripotency. Further, the treatment significantly (P < 0.05) downregulated BAX and CASP3, and upregulated GATA6 expression in compact morulae. In conclusion, IL-6 supplementation affected the in vitro development of ovine embryos in a dose- and time-dependent manner. The beneficial effect of IL-6 on the development of late-stage embryos was mediated through the changes in gene expressions associated with JAK/STAT signalling, pluripotency, apoptosis and PrE development.
白细胞介素 6（IL-6）在体外胚胎培养（IVC）的不同阶段的补充对绵羊胚胎发育和胚胎基因表达的影响进行了研究。在晚期（72-192 小时；5、10 和 25ng/ml IL-6）或整个 IVC 期间（0-192 小时；10ng/ml IL-6）将 IL-6 添加到 IVC 培养基中，以确定其对胚胎发育的影响。此外，还研究了在 IVC 的 72 小时添加 10ng/ml IL-6 对与 JAK/STAT 信号和多能性相关的基因表达的影响，以及在致密桑椹胚（补充后 1 小时）和致密桑椹胚中的凋亡和原始内胚层（PrE）发育（补充后 48 小时）。在 IVC 的晚期阶段补充 10ng/ml IL-6 与对照（21.1±1.11%）相比，显著增加了囊胚形成（35.2±1.52%），而 5ng/ml（25.9±2.98%）或 25ng/ml（16.5±0.73%）IL-6 组。相反，整个 IVC 期间添加 IL-6（10ng/ml）与对照组相比，显著降低了卵裂率（55.4±1.57%）和囊胚形成率（14.5±1.28%）（分别为 65.8±1.35%和 21.5±0.97%）。在 8-16 细胞胚胎和致密桑椹胚中，IL-6 处理显著影响与 JAK/STAT 信号和多能性相关的基因表达。此外，处理还显著下调了致密桑椹胚中的 BAX 和 CASP3，并上调了 GATA6 的表达。总之，IL-6 的补充以剂量和时间依赖的方式影响绵羊胚胎的体外发育。IL-6 对晚期胚胎发育的有益影响是通过与 JAK/STAT 信号、多能性、凋亡和 PrE 发育相关的基因表达的变化介导的。
