Fig 1.
A. Histological comparison of ovarian morphology. B. Comparison of the total number of classified follicles. C. Comparison of the total number of atretic follicles.
Compared with the control group, the number of normal follicles in POF, CH +, CH ++ and CH +++ groups was significantly decreased (* P <0.05 vs. the control group); Compared with the POF group, the number of normal follicles in HPE, CH ++ and CH +++ group was significantly increased (* P <0.05 vs. the POF group); Compared with the Control group, the number of atretic follicles in POF and CH + groups was increased significantly (*P <0.05 vs. the control group); Compared with the POF group, the number of atretic follicles in the HPE, CH ++ and CH +++ groups was significantly reduced (*P <0.05 vs. the POF group).
Fig 2.
Determination of serum hormone levels.
A. Compared with the control group, serum levels of E2 was significantly decreased in POF, CH +, CH + + groups (* P <0.05). Compared with the POF group, serum levels of E2 was significantly increased in HPE, CH++ and CH+++ groups (* P <0.05). B. Compared with the control group, serum levels of FSH was significantly increased in POF, CH +, CH + + and CH+++groups (* P <0.05). Compared with the POF group, serum levels of FSH was significantly decreased in HPE, CH++ and CH+++ groups (* P <0.05). C. Compared with the control group, serum levels of LH were significantly increased in POF, CH + and CH + + groups (* P <0.05). Compared with the POF group, serum levels of FSH was significantly decreased in HPE, CH++ and CH+++ groups (* P <0.05). D. Compared with the control group, serum levels of Progesterone was significantly decreased in POF, CH + and CH + + groups (* P <0.05). Compared with the POF group, serum levels of Progesterone were significantly increased in HPE, CH++ and CH+++ groups (* P <0.05).
Fig 3.
A. Gross morphology of mice and ovaries of each group. B. Compared with the control group, the ovarian weight of POF and CH + groups was significantly decreased, (*P<0.05). Compared with the POF group, the ovarian weight of HPE, CH++ and CH+++ groups was significantly increased (*P<0.05). C. Compared with the control group, the body weight of POF, CH + and CH++ groups was significantly decreased (*P<0.05), Compared with the POF group, the bodyweight of HPE, CH++ and CH+++ groups was significantly increased.
Fig 4.
A. Flow cytometry analysis was performed to measure the apoptotic status of granulosa cell. B. Compared with control group, the apoptotic rate of ovarian granulosa cells in POF, CH +, CH ++ and CH +++ groups was significantly higher (*P<0.05); Compared with POF group, the sum of apoptotic rate of ovarian granulosa cells in HPE, CH ++ and CH +++ groups was significantly lower (*P<0.05).
Fig 5.
HPE inhibited protein expression of p-Rictor, bad, bax, PPAR and promoted protein expression of p-AKT and p-Foxo3a in POF model mice.
A. Ovaries were harvested at day 1 after the final administration of HPE or saline, equivalent amount of whole ovary detergent lysates was western blotted with indicated antibodies. Per lane represents individual animals (5–8 animals per group). B. Compared with control group, the level of p-AKT and p-Foxo3a in POF group was significantly lower, the level of p-Rictor, Bad, Bax, PPAR in POF group was significantly higher, (*P<0.05). Compared with POF group, the level of p-Rictor, Bad, Bax, PPAR, p-AKT and p-Foxo3a in CH+, CH ++ and CH +++ groups was significantly higher (*P<0.05). Data were pooled and represented as mean ± SD, n = 5–8 animals per group.