The In Vitro Effect of Steroid Hormones, Arachidonic Acid, and Kinases Inhibitors on Aquaporin 1, 2, 5, and 7 Gene Expression in the Porcine Uterine Luminal Epithelial Cells during the Estrous Cycle

Abstract

Aquaporins (AQPs) are integral membrane proteins, which play an important role in water homeostasis in the uterus. According to the literature, the expression of aquaporins in reproductive structures depends on the local hormonal milieu. The current study investigated the effect of selected PKA kinase inhibitor H89 and MAPK kinase inhibitor PD98059, on the expression of AQP1, 2, 5, and 7, and steroid hormones (E2), progesterone (P4), and arachidonic acid (AA) in the porcine endometrium on days 18–20 and 2–4 of the estrous cycle (the follicular phase where estrogen and follicle-stimulating hormone (FSH) are secreted increasingly in preparation for estrus and the luteal phase where the ovarian follicles begin the process of luteinization with the formation of the corpus luteum and progesterone secretion, respectively). The luminal epithelial cells were incubated in vitro in the presence of the aforementioned factors. The expression of mRNA was determined by the quantitative real-time PCR technique. In general, in Experiment 1, steroid hormones significantly increased expression of AQP1, 2, and 5 while arachidonic acid increased expression of AQP2 and AQP7. On the other hand, MAPK kinase inhibitor significantly decreased the expression of AQP1 and 5. In Experiment 2, E2, P4, or AA combined with kinase inhibitors differentially affected on AQPs expression. E2 in combination with PKA inhibitor significantly decreased expression of AQP1 but E2 or P4 combined with this inhibitor increased the expression of AQP5 and 7. On the contrary, E2 with PD98059 significantly increased AQP5 and AQP7 expression. Progesterone in combination with MAPK kinase inhibitor significantly downregulated the expression of AQP5 and upregulated AQP7. Arachidonic acid mixed with H89 or PD98059 caused a decrease in the expression of AQP5 and an increase of AQP7. The obtained results indicate that estradiol, progesterone, and arachidonic acid through PKA and MAPK signaling pathways regulate the expression of AQP1 and AQP5 in the porcine luminal epithelial cells in the periovulatory period.

Oridonin alleviates lipopolysaccharide-induced acute lung injury in mice through inhibiting apoptosis, oxidative stress, and inflammation by modulating VIP/cAMP/PKA/AQPs signaling pathway

“…cAMP can bind to the regulatory subunit of PKA to promote phosphorylation of PKA, and a complete VIP/cAMP/PKA activation pathway is formed [31]. AQPs, which are closely related to the water and liquid metabolism of tissue cells, are one of the main downstream target proteins in this pathway [32]. VIP/cAMP/PKA/AQPs play a critical role in the regulation of water and liquid metabolism of lung histiocytes [33].…”

“…AQP1 is highly dependent on the phosphorylated PKA regulatory pathway. p-PKA regulates AQP1 by binding to phosphorylation sites in AQP protein, affecting water-liquid transport and cell endocytosis and exocytosis [32]. VIP/cAMP is also the activator of p-PKA, which will increase the water and liquid transport capacity of AQP1, and enhance the re ux absorption of the lymphatic and venous system, thereby reducing the interstitial lung and alveolar edema [36].…”

Time Series Ovarian Transcriptome Analyses of the Porcine Estrous Cycle Reveals Gene Expression Changes during Steroid Metabolism and Corpus Luteum Development

“…Although we previously analyzed the whole estrous cycle using RNA-seq, we focused on the interaction and connectivity between three reproductive tissues (ovary, oviduct, and endometrium), rather than on examining the changes that occurred during the estrous cycle in a particular tissue [8]. Furthermore, few studies have been carried out on the entire gene expression changes during the development of follicles and CL in the porcine estrous cycle [8,9]. In this study, we aimed to provide a comprehensive understanding of the gene expression patterns in porcine ovarian transcriptomes during the entire estrous cycle through DEG profiling, cataloging, and description of molecular mechanisms.…”