Anthraquinones in the aqueous extract of Cassiae semen cause liver injury in rats through lipid metabolism disorder.

BACKGROUND: Cassiae semen has been used as the tea or medicine component to treat hyperlipidemia or for hepatoprotection. However, Cassiae semen was reported to be a potentially hepatotoxic herb, and the underlying hepatotoxicity mechanisms or specific hepatotoxic components of Cassiae semen are unknown. PURPOSE: In this study, we aimed to explore the potential hepatotoxicity mechanisms and the hepatotoxic components of Cassiae semen. METHODS: Both young adult male and female SD rats were orally administrated with the aqueous extract of the seeds of Senna obtusifolia (L.) H.S.Irwin & Barneby at doses of 4.73, 15.75, 47.30 g/kg for 28 days, and the body weight, liver coefficient, bile acids, histopathology, serum levels of TC, TG, LDL, HDL, ALP, ALT, AST, and LDH were examined. Lipidomic analysis of rat serum was performed by LC-MS to investigate the specifically changed lipids caused by the aqueous extract treatment. The components absorbed in plasma were detected by UHPLC-Q-Exactive-MS. MTT assay was used to evaluate the cytotoxicity of these components absorbed in plasma. RESULTS: The serum levels of ALP, AST, ALT, LDH were increased on day 7 with some of which gradually dropped to normal level on day 28. In high dose of the aqueous extract treated group, the histopathological changes were observed based on the cytoplasmic vacuolation in the liver and the increase of bile acids, indicating the hepatotoxicity of the aqueous extract. The changes of TC, TG, LDL, HDL indicated the disorder of lipid metabolism. By comparing the difference in lipids between high dose group and control group, the results showed that the alterations were primarily focused on glycerophospholipid metabolism in both male and female rats. In addition, the glycerolipid metabolism in female rats also changed. Further analyses found that PC (18:2/20:4) and LysoPC 18:0 were significantly increased. Among these phytochemicals detected in plasma, nine components in the aqueous extract were considered to have the highest concentrations, particularly some types of anthraquinones (AQs) existing in Cassiae semen (AQs-in-CS), such as obtusifolin, aurantio-obtusin, and obtusin. The MTT assay showed that emodin, obtusifolin, rhein, aurantio-obtusin, and obtusin inhibited cell viability. Considering plasma concentrations and cytotoxicity of these components, our study indicates that the AQs-in-CS (obtusifolin, aurantio-obtusin and obtusin), emodin and rhein are the potential hepatotoxic phytochemicals in the aqueous extract.

Olfactomedin 1 Deficiency Leads to Defective Olfaction and Impaired Female Fertility.

Olfactomedin 1 (OLFM1) is a glycoprotein highly expressed in the brain. Olfm1(-/-) female mice were previously reported to have reduced fertility. Previous microarray analysis revealed Olfm1 among the most highly upregulated genes in the uterine luminal epithelium upon embryo implantation, which was confirmed by in situ hybridization. We hypothesized that Olfm1 deficiency led to defective embryo implantation and thus impaired fertility. Indeed, Olfm1(-/-) females had defective embryo implantation. However, Olfm1(-/-) females rarely mated and those that mated rarely became pregnant. Ovarian histology indicated the absence of corpora lutea in Olfm1(-/-) females, indicating defective ovulation. Superovulation using equine chorionic gonadotropin-human chorionic gonadotropin rescued mating, ovulation, and pregnancy, and equine chorionic gonadotropin alone rescued ovulation in Olfm1(-/-) females. Olfm1(-/-) females had a 13% reduction of hypothalamic GnRH neurons but comparable basal serum LH levels and GnRH-induced LH levels compared with wild-type controls. These results indicated no obvious local defects in the female reproductive system and a functional hypothalamic-pituitary-gonadal axis. Olfm1(-/-) females were unresponsive to the effects of male bedding stimulation on pubertal development and estrous cycle. There were 41% fewer cFos-positive cells in the mitral cell layer of accessory olfactory bulb upon male urine stimulation for 90 minutes. OLFM1 was expressed in the main and accessory olfactory systems including main olfactory epithelium, vomeronasal organ, main olfactory bulb, and accessory olfactory bulb, with the highest expression detected in the axon bundles of olfactory sensory neurons. These data demonstrate that defective fertility in Olfm1(-/-) females is most likely a secondary effect of defective olfaction.

Postweaning dietary genistein exposure advances puberty without significantly affecting early pregnancy in C57BL/6J female mice.

An epidemiological study indicates higher plasma level of genistein in girls with earlier puberty. This study tests the hypothesis in C57BL/6J mice that postweaning (peripubertal) dietary genistein exposure could result in earlier puberty in females assessed by vaginal opening, estrous cyclicity, corpus luteum and mammary gland development. Newly weaned female mice were fed with 0, 5, 100, or 500 ppm genistein diets. Decreased age at vaginal opening, increased length on estrus stage, and accelerated mammary gland development were detected in 100 and 500 ppm genistein-treated groups. Increased presence of corpus luteum was found in 5 ppm genistein-treated group at 6 weeks old only. Increased expression of epithelial-specific genes but not that of ERα or ERß was detected in 500 ppm genistein-treated mammary glands at 5 weeks old. No significant adverse effect on embryo implantation was observed. These data demonstrate causal effect of dietary genistein on earlier puberty in female mice.