Objective: In this study, we examined changes in serum leptin levels during the estrus cycle and the role of estrogen in these changes. Methods: We measured serum leptin levels during normal estrus cycles in intact rats and estradiol-17β (E₂)-induced artificial estrus cycles in ovariectomized rats. Results: Serum leptin levels increased 1.6-fold from 4.2 ± 0.2 ng/ml during diestrus stage 2 to 6.7 ± 0.9 ng/ml during proestrus stage during the 4-day estrus cycle. During the E₂-induced estrus cycle, serum leptin levels increased 2.3-fold from 2.3 ± 0.1 ng/ml at estrus to 5.4 ± 1.2 ng/ml at proestrus. E₂ also increased serum leptin concentrations and leptin mRNA expression in adipose tissue of immature rats. Discussion: These findings suggest that increased serum leptin induced by estrogen during proestrus may trigger the preovulatory release of luteinizing hormone. Furthermore, our findings indicate that estrogen has a positive effect on leptin production in adipose tissue.

Hypercholesterolemia and diabetes mellitus are known to be accompanied by reproductive dysfunction. In this study, we investigated the effects of hypercholesterolemia, hyperglycemia, and these conditions combined, on testosterone (T) and testicular luteinizing hormone/human chorionic gonadotropin (LH/hCG) binding. Sprague-Dawley rats (8 weeks old) were divided into four groups: Group 1 was the control, group 2 was fed standard chow containing 2% cholesterol (C-diet), group 3 was administered streptozotocin (STZ, 65 mg/kg, i.p.), group 4 was treated with both the C-diet and STZ. After 4 weeks, rats were sacrificed. Serum glucose was significantly higher in the STZ group (304% that of controls) and the C-diet plus STZ group (345%), but there was no difference between the C-diet group (89%) and the control group. Serum cholesterol was significantly higher in the C-diet group (206% that of controls), the STZ group (452%) and the C-diet plus STZ group (2042%). Serum T, testicular T, and LH/hCG binding were significantly lower in the C-diet group (49%, 52%, and 81% that of controls, respectively), the STZ group (15%, 32%, and 72%) and the C-diet plus STZ group (8%, 21%, and 57%). These results suggest that hypercholesterolemia is an independent risk factor for testicular dysfunction and that the reduction of serum and testicular T levels is due at least in part to a reduction in testicular LH/hCG binding in rats with hypercholesterolemia, hyperglycemia, and these conditions combined. It is further suggested that the reduction in LH/hCG binding is mainly related to a rise in serum cholesterol levels.

Abstract. The topoisomerase I inhibitor irinotecan is widely used in anticancer therapy, although the detailed mechanism is still unclear. We investigated the apoptotic mechanisms of irinotecan in human hepatocellular carcinoma (HCC) cell lines (Huh7). SN-38 caused a significant decrease in cell proliferation and induced apoptosis in Huh7 cells and HepG2 cells. SN-38 significantly increased the expression of p53 protein and its phosphorylation at Ser 15 in the nucleus and apoptosis-inducing proteins Bax, caspase-9, and caspase-3, while it significantly decreased the antiapoptosis protein Bcl-xL of Huh7 cells. SN-38-induced apoptosis was recovered after p53 antisense oligodeoxynucleotide (AS ODN) pretreatment, while Huh7 cells were precultured with p53 AS ODN, followed by the addition of SN-38 for 24 h. Furthermore, increases in p53 DNA-binding activity were observed in the nuclei of Huh7 cells after SN-38 treatment as shown by electrophoretic mobility shift analysis. SN-38 binding motifs were detected in the proximal promoter of p53 (bases −433 to −317 and −814 to −711). These results suggest that the p53-mediated apoptosis pathway is important in the anticancer effects of irinotecan in hepatocellular carcinoma.

In the rat, testosterone (T) in the neonatal period plays an important role in sexual differentiation and there is a serum T surge in male rats 2 hours after birth. Pregnant female rats were exposed to various doses of bisphenol A (BPA) from gestational day 1 (GD1) through 2 hours after parturition. About half of the BPA-exposed and control dams were subjected to cesarean section on GD22. The male fetuses on GD22 were immediately sacrificed and blood was collected. The other half of the BPA-treated and control dams delivered at GD23 (parturition day). The male pups were sacrificed 2 hours after birth. Serum T concentration was determined by radioimmunoassay (RIA). The BPA concentration in the fetal serum on GD22 increased inversely to the T levels in the serum. The T concentration in the pups' serum 2 hours after birth decreased inversely to the BPA concentration in the serum. However, there were no differences in the serum T concentration among the various doses of BPA. These results suggest that exposure to BPA in utero inhibits the T surge in the neonatal period and we speculate that exposure to BPA in utero disrupts the endocrine environment in the neonatal male.

Irinotecan, a DNA topoisomerase I inhibitor, is widely used in cancer chemotherapy. However, little is known of the mechanisms of its antitumor effects and the development of drug resistance in human hepatocellular carcinoma (HCC). In this study, we investigated the effects of short-term culture with SN-38, the active metabolite of irinotecan, on apoptosis in Huh7 cells. The cells were cultured with SN-38 for 24, 72, and 120 h, and apoptosis was determined using the terminal dUTP nick-end labeling (TUNEL) assay. The expressions of p53, apoptosis-related proteins, and P-glycoprotein (P-gp), a protein conferring the multidrug-resistant phenotype, were analyzed using Western blotting. Induced expression of P-gp was detected using fluorescence microscopy. SN-38 significantly induced apoptosis in Huh7 cells at 24 h. SN-38 also increased the expression of p53, Bax, and caspase-9 and decreased Bcl-xL expression in Huh7 cells. SN-38 decreased p53 expression and increased P-gp expression after 120 h, resulting in inhibition of apoptosis. This inhibition was reversed by the addition of verapamil to the culture medium during 120 h incubation. SN-38-induced P-gp expression was additionally enhanced by p53 decoy oligodeoxynucleotide. The changes in P-gp expression were directly moderated by p53 gene downregulation, suggesting that it plays a role in the mechanism of drug resistance. These results suggest that the accumulation of irinotecan in HCC leads to the development of drug resistance.