Astragalus polysaccharide regulates the Sirt1 / FoxO1 pathway and inhibits granulosa cell autophagy in polycystic ovary syndrome rats
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Hunan Maternal and Child Health Hospital, Changsha 410008, China

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R-33

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    Abstract:

    Objective To investigate the effect of astragalus polysaccharides (APS) on autophagy of granulosa cells in polycystic ovary syndrome ( PCOS) rats by regulating the silent mating type information regulation 2 homolog 1 (Sirt1) / forkhead box transcription factor O1 (FoxO1) pathway. Methods Ovarian granulosa cells were isolated from rats and identified by follicle stimulating hormone receptor (FSHR) immunofluorescence. CCK8 was used to assess the effect of APS on the proliferation of ovarian granulosa cells. Ovarian granulosa cells were treated with 1× 10-5 mol / L testosterone propionate and 10 μmol / L C2 ceramide for 24 h to induce the autophagy model of PCOS rat granulosa cells. CCK8 was used to measure cell proliferation. Autophagy was observed by transmission electron microscope. Protein expression of Sirt1, FoxO1, and microtubule-associated protein 1 light chain 3 ( LC3 ) was detected by Western blot. Results Immunofluorescence showed that average positive expression of FSHR protein in rat ovarian granulosa cells was 93. 18% > 90%. In 0, 100, 200, and 400 μg / mL APS-treated normal ovarian granulosa cells, there was no significant difference in OD450 after adherence at 0 or 24 h (P > 0. 05). Compared with the normal group, a large number of autophagy bodies appeared near the nucleus of the model group, and double and single layer membranes wrapped the cytoplasm to form a closed and round structure. Some of the autophagy body inner membrane had dissolved and there was a single membrane structure around the autophagosome, which contained some degraded cytoplasm similar to the autophagosome structure. The cell structure of the 100 μg / mL APS group was similar to that of the model group. With the increase of APS dosage, the number of autophagy bodies decreased and the cells were normal in the 400 μg / mL APS group. There was no significant difference in OD450 between normal, model, and 100, 200, and 400 μg / mL APS groups at 0 h of cell adhesion (P> 0. 05). After 24 h of cell adhesion, compared with the normal group, OD450 was lower in the model group (P< 0. 05) and the protein levels of Sirt1, FoxO1, and LC3II/ LC3I were higher (P< 0. 05). Compared with the model group, OD450 was higher in 100, 200, and 400 μg / mL APS groups (P< 0. 05) and the protein levels of Sirt1, FoxO1, and LC3II/ LC3I were lower (P< 0. 05). Conclusions APS inhibits autophagy of granulosa cells in PCOS rats, which may be mediated by inhibiting the autophagy pathway Sirt1 / FoxO1.

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History
  • Received:November 18,2020
  • Revised:
  • Adopted:
  • Online: November 29,2021
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