动物造模,抑制多囊卵巢综合征 z-bio 2022-03-23 325

　　Objective To investigate the effect of astragalus polysaccharide (APS) regulating the silent information regulator 1/forkhead transcription factor O1 ( Sirt1 / FoxO1) pathway on autophagy in granulosa cells of polycystic ovary syndrome (PCOS) rats.

　　Methods Ovarian granulosa cells were isolated from rats and identified by follicle-stimulating hormone receptor (FSHR) immunofluorescence, and the effect of APS on the proliferation of ovarian granulosa cells was detected by CCK8. 10-5 mol/L testosterone propionate and 10 μmol/L C2-ceramide acted on rat ovarian granulosa cells for 24 h to induce PCOS rat granulosa cell autophagy model, CCK8 was used to detect cell proliferation; transmission electron microscopy was used to observe autophagosomes; The expression of Sirt1, FoxO1 and microtubule-associated protein 1 light chain 3 (LC3) protein in cells was detected by western blotting.

　　Results Immunofluorescence detection showed that the average positive expression of FSHR protein in rat ovarian granulosa cells was 93.18% > 90%, and the following experiments could be carried out. Compared with normal ovarian granulosa cells treated with 0, 100, 200, and 400 μg/mL APS, there was no significant difference in the OD450 of cells at 0 and 24 h after adherence (P>0.05). Compared with the normal group, a large number of autophagosomes appeared near the nucleus of the model group, double-layer and single-layer membrane wrapped the cytoplasm to form a closed, round structure, some autophagosome inner membrane was dissolved, and a monolayer was seen around the autophagosome. The membrane structure wrapped some degraded cytoplasm, similar to autophagosome structure; the 100 μg/mL APS group had similar cell structure to the model group. Cells were normal in the mL APS group. There was no significant difference in cell OD450 between normal group, model group, 100, 200, and 400 μg/mL APS group at 0 h after cells adhered (P>0.05). After the cells adhered for 24 h, compared with the normal group, the OD450 of the cells in the model group decreased (P<0.05), and the protein levels of Sirt1, FoxO1, LC3II/LC3Ⅰin the cells increased (P<0.05). Compared with the 100, 200, and 400 μg/mL APS groups, the OD450 of the cells was increased (P<0.05), and the protein levels of Sirt1, FoxO1, LC3II/LC3Ⅰin the cells were decreased (P<0.05).

　　Conclusion APS can inhibit the autophagy of granulosa cells in PCOS rats, which may be achieved by inhibiting the autophagy pathway Sirt1/FoxO1.