Figure 2
HA protects the immunosuppressive agent-induced damage to GCs in vitro.
(a) After treatment with HA (0, 100, 200 and 500 μg/ml) for 24 h, 36 h and 48 h, cell viability of KGN was tested using a CCK8-kit. *P < 0.05, **P < 0.01 compared with the cells not treated with HA. (b) KGN cells were treated with CYC (100 μM) or TG (50 nM) in the presence or absence of HA (200 μg/ml) for 24 h, 36 h and 48 h respectively. The cell proliferation was analyzed using a CCK8-kit. *P < 0.05 vs. control; #P < 0.05 CYC+HA vs. CYC; &P < 0.05 TG+HA vs. TG. (c) The proliferation of HA-treated KGN cells was analyzed using the CFSE staining method (upper panel). The data were normalized to the proliferative index (PI) of control cells (lower panel). *P < 0.05 compared with control. (d) Primary GCs were obtained from rat ovary and an optical microscope image was taken (×60). (e) Rat GCs cultured on coverslips were fixed and immunostained with specific antibodies for rat FSHR (×60). (f) After treatment with the indicated concentration of HA for 48 h, rat GC viability was analyzed using CCK8 kit. **P < 0.01vs control. (g) Rat primary GCs were treated with CYC (100 μM) or TG (50 nM) in the presence or absence of HA (200 μg/ml) for 24 h, 36 h and 48 h respectively. The cell viability was analyzed using a CCK8-kit. *P < 0.05 vs. control; #P < 0.05 CYC+HA vs. CYC; &P < 0.05 TG+HA vs. TG. (h) Primary GCs were treated with CYC (100 μM) or TG (50 nM) in the presence or absence of HA (200 μg/ml) for 48 h. The concentration of E2 and P4 was assessed using ELISA. #P < 0.05 CYC+HA vs. CYC; $P < 0.05 TG+HA vs. TG. Average results from three independent experiments are shown.
