School of Public Health, Shandong University, 44 Wenhua Xi Road, Jinan, Shandong, 250012, PR China.
School of Public Health, Shandong University, 44 Wenhua Xi Road, Jinan, Shandong, 250012, PR China.
Chem Biol Interact. 2019 Oct 1;312:108751. doi: 10.1016/j.cbi.2019.108751. Epub 2019 Jul 29.
To explore the effects of carbon disulfide (CS) and N-carbamoyl glutamate (NCG) on autophagy during the window of embryo implantation in mice and whether dietary NCG supplementation can promote embryo implantation in case of CS exposure.
Pregnant mice that received single intraperitoneal injection of CS on Gestational day (GD)4 were fed basal diet with or without NCG supplementation from GD1 to endpoints. The control mice were injected solvents. There were four endpoints (GD5, GD6, GD7 and GD9 endpoints) in each group. The uterus was collected on endpoints to detect autophagy-related markers by using the methods of transmission electron microscopy (TEM), immunohistochemistry (IHC), quantitative real-time polymerase chain reaction (qRT-PCR) and ELISA.
The P62 brown punctate staining increased in CS exposure group and reduced after dietary NCG supplementation, which was opposite with LC3B, Beclin1 and ATG5 on GD5 endpoint. Simultaneously, P62 protein expression raised 43.33% on GD5 endpoint (p < 0.01) when exposed to CS and descended to the control level after NCG supplementation. The rate of decline of LC3B and Beclin1 proteins were 27.04% (p < 0.01) and 23.27% (p < 0.05) on GD5 endpoint, 20.20% (p < 0.05) and 11.30% on GD7 endpoint in CS exposure group, respectively, then NCG supplementation caused the LC3B and Beclin1 protein expression to rise in different degrees. Comparatively, the mRNA expression of all autophagy-related gene changed more apparently on three endpoints than the protein expression. The images of TEM showed that nearly no autophagosome could be seen in CS exposure group, while dietary NCG supplementation increased the number of autophagosome obviously on GD5 endpoint. The number of implanted embryos which declined due to CS exposure returned to normal in NCG supplementation group.
Dietary NCG supplementation could rescue the suppressed autophagy induced by CS in the window of implantation and increase the number of implanted embryos.
探讨二硫化碳（CS）和 N- 氨甲酰谷氨酸（NCG）在小鼠胚胎着床窗口期对自噬的影响，以及在 CS 暴露的情况下，膳食 NCG 补充是否能促进胚胎着床。
妊娠第 4 天（GD4）给小鼠单次腹腔注射 CS，从 GD1 到终点给小鼠喂食基础饲料或补充 NCG。对照组小鼠注射溶剂。每组有 4 个终点（GD5、GD6、GD7 和 GD9 终点）。在终点时收集子宫，使用透射电子显微镜（TEM）、免疫组织化学（IHC）、实时定量聚合酶链反应（qRT-PCR）和 ELISA 方法检测自噬相关标志物。
CS 暴露组 P62 棕色点状染色增加，补充膳食 NCG 后减少，这与 GD5 终点时的 LC3B、Beclin1 和 ATG5 相反。同时，CS 暴露组 P62 蛋白表达在 GD5 终点时升高了 43.33%（p<0.01），补充 NCG 后降至对照水平。LC3B 和 Beclin1 蛋白在 GD5 终点的下降率分别为 27.04%（p<0.01）和 23.27%（p<0.05），CS 暴露组在 GD7 终点时分别为 20.20%（p<0.05）和 11.30%（p<0.05），然后 NCG 补充使 LC3B 和 Beclin1 蛋白表达在不同程度上升高。相比之下，所有自噬相关基因的 mRNA 表达在三个终点都比蛋白表达变化更明显。TEM 图像显示，CS 暴露组几乎看不到自噬体，而膳食 NCG 补充在 GD5 终点明显增加了自噬体的数量。由于 CS 暴露而减少的着床胚胎数量在 NCG 补充组恢复正常。
膳食 NCG 补充可以挽救 CS 诱导的着床窗口期抑制的自噬，并增加着床胚胎的数量。
