The effect of diet and cardiovascular risk on ovarian aging in cynomolgus monkeys (Macaca fascicularis).

OBJECTIVE: The aim of this study was to determine the relationships among dietary protein source, cardiovascular risk, reproductive hormones, and ovarian aging. METHODS: Adult female cynomolgus monkeys (Macaca fascicularis) were assigned randomly to one of two diets containing saturated fat and cholesterol, differing only by protein source: (1) casein-lactalbumin (n = 29) or (2) soy protein with isoflavones (n = 32). Cardiovascular risk markers and reproductive hormones were measured at baseline and after 32 months of treatment, at which time the ovaries were removed and serially sectioned and ovarian follicles were counted in every 100th section. RESULTS: Casein-lactalbumin-fed monkeys had fewer primordial, primary, and secondary follicles (all P values < 0.05) than did their soy-fed counterparts. Antimüllerian hormone was significantly correlated with all follicle types (r values > or = 0.66, P < 0.001) for casein-fed monkeys and was significantly correlated with primary (rsoy = 0.47, P = 0.005) and secondary (rsoy = 0.45, P = 0.007) follicles in soy-fed monkeys. No significant associations were seen between any of the other reproductive hormones measured and follicle counts. Casein-lactalbumin-fed monkeys had a more atherogenic lipoprotein profile and increased atherosclerosis extent (P < 0.05), but despite these differences in cardiovascular risk between monkeys fed with casein-lactalbumin and monkeys fed with soy, none of the individual cardiovascular risk markers measured in this study explained the relationship between dietary protein source and follicle counts (linear regression, all P values > 0.05). CONCLUSIONS: Diet influences the rate of follicular depletion in cynomolgus macaques; however, the mechanism for this effect remains undetermined.

Experimental induction of reduced ovarian reserve in a nonhuman primate model (Macaca fascicularis).

Chronic diseases including coronary heart disease and osteoporosis represent a substantial health burden to postmenopausal women, yet the initiation of these conditions and their relationships with reproductive aging remain poorly understood. This situation is due, in part, to the lack of animal models reflecting ovarian and hormonal characteristics of peri- and postmenopausal women. Ovaries of women approaching menopause are nearly depleted of primordial follicles but retain a pool of larger developing follicles and androgen-producing stroma, a condition known as reduced ovarian reserve (ROR). The long-term goal of the research presented here was to create a monkey model of reproductive aging, beginning with ROR and progressing to perimenopause and finally postmenopause. Here we sought to develop a method to reduce primordial follicles in cynomolgus monkeys (Macaca fascicularis) and document hormonal changes associated with follicle reduction or ROR. At 30 d after surgical placement of a biodegradable fiber containing approximately 200 mg of 4-vinlycyclohexene diepoxide (VCD) next to one ovary in each of 8 monkeys, primordial follicles were reduced by approximately 70%, with a corresponding decrease (83%) in antimüllerian hormone (AMH, a serum marker of ovarian follicle numbers). At 4 mo after VCD-treatment of both ovaries in 29 monkeys (approximately 200 mg VCD per ovary), AMH was reduced 56% from baseline, testosterone was unchanged, and follicular phase estradiol was slightly increased. These data indicate that VCD treatment markedly reduced primordial follicles while preserving larger estradiol- and testosterone-producing follicles and ovarian stroma, a condition that mimics ROR in women.

Effects of 4-vinylcyclohexene diepoxide on peripubertal and adult Sprague-Dawley rats: ovarian, clinical, and pathologic outcomes.

Young rats treated daily with intraperitoneal 4-vinylcyclohexene diepoxide (VCD) undergo selective destruction of primordial follicles, resulting in gradual ovarian failure resembling the menopausal transition in women. To determine whether VCD has similar effects on ovaries of older rats, adult and peripubertal Sprague-Dawley rats were injected intraperitoneally daily for 30 d with vehicle or VCD at 40 or 80 mg/kg. Body weight, food intake, complete blood counts, and markers of liver injury and renal function were measured during VCD treatment. Complete gross necropsy and microscopic observations were performed on day 31, and ovarian follicles were counted. At 80 mg/kg, VCD destroyed primordial and primary follicles to a similar extent in both adult and peripubertal animals, although adult rats likely started with fewer follicles and therefore approached follicle depletion. Treatment with VCD did not affect body weight, but food intake was reduced in both adult and peripubertal rats treated with 80 mg/kg VCD. Adult rats treated with 80 mg/kg VCD had neutrophilia and increased BUN and creatinine; in addition, 4 of these rats were euthanized on days 25 or 26 due to peritonitis. VCD treatment did not increase alanine aminotransferase levels, a marker of liver injury, although the 80-mg/kg dose increased liver weights. In conclusion, VCD effectively destroys small preantral follicles in adult Sprague-Dawley rats, making them a suitable model of the menopausal transition of women. However, because adult rats were more sensitive to the irritant properties of VCD, the use of a lower dose should be considered.