Guo Yuli, Liu Junping, Zhang Shuangyi, Sun Di, Dong Zhiying, Cao Jinshan
Key Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China.
Vet Sci. 2024 Dec 17;11(12):663. doi: 10.3390/vetsci11120663.
(AMK) as an oriental medicine has been used in the treatment of threatened abortion. (AT-I) is one of the major bioactive components of AMK. This study aimed to investigate the effect of AT-I on the secretion of estradiol (E) and progesterone (P) in feline ovarian granulosa cells (FOGCs), which is necessary for pregnancy. At first, the proliferation of FOGCs after AT-I treatment was measured by CCK-8. Then, the synthesis of E and P were measured by ELISA. Lastly, transcriptome sequencing was used to detect the DEGs in the FOGCs, and RNA-seq results were verified by RT-qPCR and biochemical verification. It was found that AT-I could promote proliferation and the secretion of E and P in FOGCs; after AT-I treatment, 137 significant DEGs were observed, out of which 49 were up-regulated and 88 down-regulated. The DEGs revealed significant enrichment of 52 GO terms throughout the differentiation process ( < 0.05), as deciphered by Gene Ontology enrichment analysis. Kyoto Encyclopedia of Genes and Genomes analysis manifested that the DEGs were successfully annotated as members of 155 pathways, with 23 significantly enriched ( < 0.05). A relatively high number of genes were enriched for the cholesterol metabolism, ovarian steroidogenesis, and biosynthesis of unsaturated fatty acids. Furthermore, the contents of the total cholesterol and low-density lipoprotein cholesterol were decreased by AT-I treatment in the cell culture supernatant. The results indicated that AT-I could increase the ability of FOGCs to secrete E and P, which might be achieved by activation of cholesterol metabolism.
当归芍药散（AMK）作为一种传统中药，已被用于治疗先兆流产。阿魏酸-Ⅰ（AT-I）是AMK的主要生物活性成分之一。本研究旨在探讨AT-I对猫卵巢颗粒细胞（FOGCs）中雌二醇（E）和孕酮（P）分泌的影响，这两种激素对妊娠至关重要。首先，通过CCK-8法检测AT-I处理后FOGCs的增殖情况。然后，采用ELISA法检测E和P的合成。最后，利用转录组测序检测FOGCs中的差异表达基因（DEGs），并通过RT-qPCR和生化验证对RNA-seq结果进行验证。结果发现，AT-I可促进FOGCs的增殖以及E和P的分泌；AT-I处理后，共观察到137个显著的DEGs，其中49个上调，88个下调。通过基因本体富集分析可知，这些DEGs在整个分化过程中显著富集了52个GO术语（P<0.05）。京都基因与基因组百科全书分析表明，这些DEGs成功注释为155条通路的成员，其中23条显著富集（P<0.05）。相对较多的基因富集于胆固醇代谢、卵巢类固醇生成和不饱和脂肪酸生物合成。此外，AT-I处理可降低细胞培养上清液中总胆固醇和低密度脂蛋白胆固醇的含量。结果表明，AT-I可提高FOGCs分泌E和P的能力，这可能是通过激活胆固醇代谢实现的。
