Laboratory of Metabolic Manipulation of Herbivorous Animal Nutrition, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China.
Int J Mol Sci. 2024 Nov 19;25(22):12428. doi: 10.3390/ijms252212428.
The circadian gene is recognized for its regulatory effects on cell proliferation and lipid metabolism across various non-ruminant cells. This study investigates the influence of gene overexpression on goat rumen epithelial cells using a constructed pcDNA3.1- plasmid, assessing its impact on circadian gene expression, cell proliferation, and mRNA levels of short-chain fatty acid (SCFA) transporters, alongside genes related to lipid metabolism, cell proliferation, and apoptosis. Rumen epithelial cells were obtained every four hours from healthy dairy goats (n = 3; aged 1.5 years; average weight 45.34 ± 4.28 kg), cultured for 48 h in vitro, and segregated into control (pcDNA3.1) and overexpressed (pcDNA3.1-) groups, each with four biological replicates. The study examined the potential connection between circadian rhythms and nutrient assimilation in ruminant, including cell proliferation, apoptosis, cell cycle dynamics, and antioxidant activity and the expression of circadian-related genes, VFA transporter genes and regulatory factors. The introduction of the pcDNA3.1- plasmid drastically elevated expression levels by 3471.48-fold compared to controls ( < 0.01), confirming effective overexpression. overexpression resulted in a significant increase in apoptosis rates ( < 0.05) and a notable reduction in cell proliferation at 24 and 48 h post-transfection ( < 0.05), illustrating an inhibitory effect on rumen epithelial cell growth. elevation significantly boosted the expression of , , , and ( < 0.05) while diminishing expression ( < 0.05). While the general expression of intracellular inflammation genes remained stable, expression notably increased. Antioxidant marker levels (SOD, MDA, GSH-Px, CAT, and T-AOC) exhibited no significant change, suggesting no oxidative damage due to overexpression. Furthermore, overexpression significantly downregulated , , , and mRNA expressions while upregulating and . These results suggest that overexpression impairs cell proliferation, enhances apoptosis, and modulates VFA transporter-related factors in the rumen epithelium. This study implies that the gene may regulate VFA absorption through modulation of VFA transporters in rumen epithelial cells, necessitating further research into its specific regulatory mechanisms.
该节律基因在多种非反刍细胞中对细胞增殖和脂质代谢具有调节作用。本研究通过构建 pcDNA3.1-质粒，研究基因过表达对山羊瘤胃上皮细胞的影响，评估其对节律基因表达、细胞增殖以及短链脂肪酸（SCFA）转运体 mRNA 水平的影响，以及与脂质代谢、细胞增殖和细胞凋亡相关的基因。从健康奶牛（n = 3；年龄 1.5 岁；平均体重 45.34 ± 4.28 kg）中每隔 4 小时获得瘤胃上皮细胞，在体外培养 48 h，并分为对照组（pcDNA3.1）和过表达组（pcDNA3.1-），每组 4 个生物学重复。该研究探讨了包括细胞增殖、凋亡、细胞周期动态、抗氧化活性和与昼夜节律相关基因、VFA 转运体基因和调节因子的表达在内的，节律与反刍动物营养吸收之间的潜在联系。pcDNA3.1-质粒的引入使表达水平比对照组高出 3471.48 倍（<0.01），证实了有效过表达。过表达导致凋亡率显著增加（<0.05），转染后 24 和 48 h 细胞增殖显著减少（<0.05），表明对瘤胃上皮细胞生长具有抑制作用。过表达显著促进了（<0.05）、（<0.05）、（<0.05）和（<0.05）的表达，同时降低了（<0.05）的表达。虽然细胞内炎症基因的总体表达保持稳定，但（<0.05）的表达显著增加。抗氧化标志物水平（SOD、MDA、GSH-Px、CAT 和 T-AOC）没有明显变化，表明过表达不会导致氧化损伤。此外，过表达显著下调（<0.05）、（<0.05）、（<0.05）和（<0.05）的 mRNA 表达，同时上调（<0.05）和（<0.05）的表达。这些结果表明，过表达会损害细胞增殖，增强凋亡，并调节瘤胃上皮细胞中 VFA 转运体相关因子。本研究表明，节律基因可能通过调节瘤胃上皮细胞中 VFA 转运体来调节 VFA 的吸收，因此需要进一步研究其具体的调节机制。
