Fig. 8

Inhibition of apoptosis of ovarian granulosa cells in PCOS rats through LncMEG3-mediated regulation of miR-21-3p after acupuncture intervention. A Experimental strategy for acupuncture intervention; B Cleaved caspase-3, caspase-3, Bax, and Bcl-2 protein bands in granulosa cells of PCOS rat ovaries after acupuncture intervention. C Protein blotting for cleaved caspase-3/caspase-3 protein expression in granulosa cells of PCOS rats after acupuncture intervention. D Western blotting for Bcl-2/Bax protein expression in granulosa cells of PCOS rats after needle prick intervention. E Expression of LncMEG3 in ovarian granulosa cells of PCOS rats detected by RT‒qPCR after acupuncture intervention. F Expression of miR-21-3p in ovarian granulosa cells of PCOS rats detected by RT‒qPCR after acupuncture intervention; G Detection of caspase-3 by RT‒qPCR in ovarian granulosa cells of PCOS rats; H Detection of Bax by RT‒qPCR in ovarian granulosa cells of PCOS rats; I Detection of Bcl-2 by RT‒qPCR in ovarian granulosa cells of PCOS rats; B The JC-1 method was used to detect the early stage of apoptosis in PCOS rat ovarian granulosa cells after MEG3 downregulation (scale bar = 100 μm): at high mitochondrial membrane potential, which can produce red fluorescence, at low mitochondrial membrane potential, which produces green fluorescence, and the blue fluorescent DAPI dye labels the nucleus, and Merge is a superimposed image showing the three fluorescent signals; C The ratio of JC-1 aggregate (red) to monomer (green) intensity in myotubes, Cell experiments were independently repeated three times; L Apoptosis of ovarian granulosa cells in PCOS rats after acupuncture intervention was detected by the TUNEL method (scale bar = 100 μm), with green fluorescent FITC dye labelling the apoptotic cells and blue fluorescent DAPI dye labelling the nuclei; the merged image is a superimposed image of both fluorescent signals; M Fluorescence values of ovarian granulosa cells in PCOS rats after acupuncture intervention reflecting apoptosis. B-M, N = 6. These data are expressed as the mean ± standard deviation, ***P < 0.0001 compared to the normal group and ^####P < 0.0001 compared to the sh-NC group, using one-way ANOVA and Tukey's post hoc test for statistical analysis. 1: Normal group, 2: PCOS group, 3: PCOS + A group, 4: PCOS + SA group, 5: PCOS + W group