Fig. 3

NPY reduced TUNEL-positivity of granulosa cells of DHT-treated rats, whereas NPY did not affect proliferation of granulosa cells. Granulosa cells were isolated from in vivo DHT treatment model rats, incubated with 10% serum overnight and analyzed Ki67 positivity for proliferation (A) and TUNEL assay for apoptosis (B). Representative pictures are shown A, B Isolated granulosa cells from in vivo DHT treatment model rats were starved for 24 h and treated with NPY (0.1 nM) in vitro for 24 h. Their Ki67 and TUNEL positivity were analyzed C, D DHT and NPY treatment did not affect Ki67 positivity. NPY in vitro treatment did not change TUNEL-positivity in CTL rats, but reduced that in DHT-treated rats. These results suggested hyperandrogenism change the NPY-mediated apoptosis in granulosa cells. Results are expressed as mean ± SEM of independent replicates (A,B; CTL, n = 6; DHT, n = 6) (C,D; CTL, n = 3; DHT, n = 3) and analyzed by unpaired t test (A,B) and two-way ANOVA following multiple comparison, Bonferroni (C,D). *, p < 0.05 (vs. with DHT, in vivo). **, p < 0.05 (vs. with DHT, in vivo, without NPY, in vitro)