Figures & data
Table 1. qRT-PCR primers.
Figure 1. HOTAIR downregulation regulated the weights of the body and ovary as well as the levels of serum hormones in PCOS rats. (A) The changes in body weight in rats; (B) Photographs of ovaries and the changes of ovarian weight in rats; (C) The expression of HOTAIR mRNA in the ovarian tissues among the rats. (D) The levels of serum hormones in rats. Unless otherwise specified, all results were presented as mean ± SD. +p < .05, and ++p < .01 vs.Control; *p < .05, and **p < .01 vs. Model; #p < .05, and ##p < .01 vs. si-HOTAIR in all figures. Note: PCOS, polycystic ovarian syndrome; TT, testosterone; E2, estradiol; LH, luteinizing hormone; FSH, follicle-stimulating hormone.
Figure 2. HOTAIR downregulation alleviated abnormal estrus cycle of the PCOS rats. The estrus cycle of the rats was evaluated by HE staining. Magnification, ×200. Note: P, proestrus; E, estrus; M, metestrus; DI, diestrus.
Figure 3. HOTAIR downregulation improved the ovarian morphology of the PCOS rats. The ovarian morphology of the rats was observed by HE staining, and the number of total follicle, preantral follicle, antral follicle, atretic follicle as well as ovarian vacuole were also counted under microscope. PF, preantral follicle; AF, atretic follicle; OV, ovarian vacuole; antral follicle was indicated by the red arrow. Magnification, ×200 and ×400.
Figure 4. HOTAIR downregulation inhibited the apoptosis of ovarian tissues in PCOS rats. A: The cell apoptosis of the ovarian tissues in the rats was detected by TUNEL. Magnification, ×200. B: Bcl-2, Bax, and Bad protein expressions of the ovarian tissues in the rats were measured by western blotting.
Figure 5. HOTAIR downregulation inhibited IGF-1 expression and PI3K/Akt pathway in PCOS rats. The expression of IGF-1 protein as well as the phosphorylation of PI3K and Akt in rats’ ovarian tissues was tested by western blotting.
Figure 6. HOTAIR downregulation elevated viability and inhibited apoptosis in ovarian GCs of PCOS. A: The expression of HOTAIR mRNA in ovarian GCs was assessed by qRT-PCR. B: The cell viability of ovarian GCs was evaluated by CCK-8. C: The apoptosis of ovarian GCs was measured by flow cytometry. D: The Bcl-2, Bax, and Bad protein expressions of the ovarian GCs were detected by western blotting. Note: GCs, granulosa cells.
Figure 7. HOTAIR downregulation inhibited IGF-1 protein expression and PI3K/Akt pathway in ovarian GCs of PCOS. The expression of IGF-1 protein as well as the phosphorylation of PI3K and Akt in ovarian GCs was tested by western blotting.
Figure 8. HOTAIR downregulation improved PCOS in vitro via IGF-1-mediated PI3K/Akt pathway. A: The viability of the ovarian GCs was tested by CCK-8. B: The apoptosis of the ovarian GCs was detected by flow cytometry. C: The Bcl-2, Bax, and Bad protein expressions of the ovarian GCs were detected by western blotting.
Figure 9. The role of LY294002 and IGF-1 on IGF-1 protein expression and PI3K/Akt pathway. The expression of IGF-1 protein as well as the phosphorylation of PI3K and Akt was measured by western blotting.
Data availability statement
All the data are contained in the manuscript.