MiR-26a promotes apoptosis of porcine granulosa cells by targeting the 3β-hydroxysteroid-Δ24-reductase gene

Abstract

Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (<i>DHCR24</i>) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs.Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and <i>DHCR24</i>-siRNA <i>in vitro</i>. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzymelinked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and <i>DHCR24</i> 3′-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the <i>DHCR24</i> mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor.Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of <i>DHCR24</i> also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within <i>DHCR24</i> 3′-UTR. Up-regulation of miR-26a inhibited <i>DHCR24</i> mRNA and protein expression in pGCs.Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of <i>DHCR24</i> in pGCs.

microRNA-26a shuttled by extracellular vesicles secreted from adipose-derived mesenchymal stem cells reduce neuronal damage through KLF9-mediated regulation of TRAF2/KLF2 axis

“…After 72 h of reperfusion, the mice were euthanized for analysis. The dose of oe-KLF2 vector was 100 nM, and n = 8 for mice in each group [47]. *, p < 0.05, vs. EVs-inhibitor NC or oe-NC or EVs-inhibitor NC + oe-NC in mouse brain tissue slices, #, p < 0.05, vs. EVs-miR-26a inhibitor + oe-NC in mouse brain tissue slices.…”

Transcriptional analysis of deoxynivalenol‐induced apoptosis of sow ovarian granulosa cell

“…Fresh porcine ovaries were obtained from a commercial slaughterhouse and transported back to the laboratory within 1 hr. Porcine ovarian granulosa cells (pGCs) were collected from porcine ovarian follicles (3–6 mm diameter) (Zhang et al, ), washed and centrifuged (1,000 g , 5 min), and then inoculated in a 6‐cm dish at a density of 1 × 10 6 cells. Cells were cultured in DMEM and Nutrient Mixture F‐12 medium (DMEM/F‐12), containing 10% FBS, 100 units/mL penicillin and 100 mg/ml streptomycin at 37°C in 5% CO 2 .…”

circSLC41A1 Resists Porcine Granulosa Cell Apoptosis and Follicular Atresia by Promoting SRSF1 through miR-9820-5p Sponging

“…However, SRSF1 function and its regulatory mechanism are rarely studied in reproduction, especially during GC apoptosis and follicular atresia. Interestingly, our previous RNAseq study highlighted a higher expression of SRSF1 in HFs than AFs, which hinted at its regulatory potential in GC apoptosis [4].…”

“…As a major component of the follicle, granulosa cells (GCs) function by closely interacting with the oocyte and theca cells and producing paracrine substances. Research has demonstrated that GCs' proliferation and function are essential in follicle development and maturation, while the GC apoptosis rate increases significantly with the progression of follicular atresia [4].…”