phenanthrenes and Infertility--Male

Eucommia ulmoides has been used for many years as a successful strategy to treat male infertility. Aucubin (AU) is the active ingredient extracted from Eucommia ulmoides. However, its protective action and exact mechanism on testicular injury is not yet known.. Here, the protective effect and the mechanism of action of AU on testis damage under oxidative stress was investigated in vivo and in vitro.. As regard the in vivo experiment, male mice were divided into five groups and testicular injury model was established by Triptolide (TP) (120 μg/kg) intraperitoneal injection for two weeks. Animals in the treatment group were pretreated with an intraperitoneal injection of AU at different doses (5, 10 and 20 mg/kg) for 1 h and subsequently treated with TP (120 μg/kg). At the end of the experimental period, the testis was collected for biochemical and histological examination. As regard the in vitro experiment, Sertoli cells (SCs) were used to investigate the protective effect and mechanism of action of AU against disruption of the blood-testis-barrier (BTB) and apoptosis induced by TP via apoptosis detection, western blot, immunofluorescence analysis, and siRNA transient transfection.. TP-treated animals showed testicular atrophy, BTB disruption, increased ROS levels and spermatogenic dysfunction. Pre-administration of AU resulted in a significant protection on keeping a normal testicular weight, sperm morphology, BTB integrity, and a normal level of oxidative stress markers and antioxidants. Furthermore, AU prevented apoptosis through an effective inhibition of PERK/CHOP and JNK dependent apoptosis pathway, as well as protected the integrity of BTB by up-regulating the expression of tight junction proteins (ZO-1, Occludin, Claudin-11) and gap junction protein (Cx43). The mechanistic study revealed that AU significantly triggered Nrf2 translocation, thus increasing nuclear Nrf2 accumulation and then induced antioxidant enzymes expression in the testis and SCs. Furthermore, Nrf2 silencing unsuccessfully reversed the increased CHOP and p-JNK expression induced by TP, abolishing the protective effect of AU.. These results indicate that AU might be considered as a potential protective agent against testicular injury.

Topics: Animals; Antioxidants; Apoptosis; Blood-Testis Barrier; Cell Line; Diterpenes; Epoxy Compounds; Eucommiaceae; Humans; Infertility, Male; Injections, Intraperitoneal; Iridoid Glucosides; Male; Mice; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Sertoli Cells; Testis; Up-Regulation

Triptolide (TP), derived from the medicinal plant Triterygium wilfordii Hook. f. (TWHF), is a diterpene triepoxide with variety biological and pharmacological activities. However, TP has been restricted in clinical application due to its narrow therapeutic window especially in reproductive system. During spermatogenesis, Sertoli cell cytoskeleton plays an essential role in facilitating germ cell movement and cell-cell actin-based adherens junctions (AJ). At Sertoli cell-spermatid interface, the anchoring device is a kind of AJ, known as ectoplasmic specializations (ES). In this study, we demonstrate that β-actin, an important component of cytoskeleton, has been significantly down-regulated after TP treatment. TP can inhibit the expression of Rho GTPase such as, RhoA, RhoB, Cdc42 and Rac1. Downstream of Rho GTPase, Rho-associated protein kinase (ROCKs) gene expressions were also suppressed by TP. F-actin immunofluorescence proved that TP disrupts Sertoli cells cytoskeleton network. As a result of β-actin down-regulation, TP treatment increased expression of testin, which indicating ES has been disassembled. In summary, this report illustrates that TP induces cytoskeleton dysfunction and disrupts cell-cell adherens junctions via inhibition of Rho GTPases.

Topics: Actins; Adherens Junctions; Animals; Diterpenes; Epoxy Compounds; Germ Cells; Infertility, Male; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley; rho GTP-Binding Proteins; Sertoli Cells

As the unique quality control standard of Tabellae Glucosidorum Tripterygii Totorum, triptolide (CAS 38748-32-2) has a narrow therapeutic window. A significant side-effect of triptolide is its male reproductive toxicity the mechanism of which is still unknown. Therefore, in the present study the male reproductive toxicity and toxicokinetics of triptolide were investigated. Male Sprague-Dawley (SD) rats were treated with triptolide by oral administration (gastric infusion; 0, 100, 200, 400 microg/kg) once daily for 8 weeks. At the end of the treatment, the concentrations of triptolide in blood and testis samples were analyzed with liquid chromatography/mass spectrometry (LC/MS) to obtain the toxicokinetic parameters. Triptolide showed a non-linear kinetics profile and was rapidly absorbed but relatively slowly eliminated in the rats. Specifically, an accumulation of triptolide was seen in the testis. In the hematological study, mean corpuscular hemoglobin concentration (MCHC) had a marginal decrease in all triptolide treated groups. Alkaline phosphatase (ALP) in serum increased significantly only in the 400 microg/kg group during clinical chemistry assays although no histopathological change was found in the hematopoietic system or liver. In the male reproductive toxicity studies, the testis and epididymis weights of all triptolide treatment groups decreased significantly. The cauda epididymis sperm content and motility even decreased to zero. Evident changes were observed in the seminiferous tubules and the epididymides of triptolide treated rats (e.g., intraepithelial vacuoles of varying sizes; increased germ cells de generation, exfoliation and tubular atrophy). These findings provide valuable information to estimate the reproductive risk of triptolide in humans.

Topics: Animals; Area Under Curve; Blood Cell Count; Body Weight; Chromatography, High Pressure Liquid; Diterpenes; Epididymis; Epoxy Compounds; Half-Life; Immunosuppressive Agents; Infertility, Male; Male; Mass Spectrometry; Organ Size; Phenanthrenes; Rats; Rats, Sprague-Dawley; Sperm Count; Sperm Motility; Testis

To observe the effects of chlorotriptolide (T4) and triptonide (T7) on the chromosome aberration and micronuclei rates of bone marrow cell in male SD rats.. Antifertility doses of T4[80 micrograms/(kg.d)] or T7[317 micrograms/(kg.d)] were given to male rats per OS for 10 weeks. Bone marrow slides were then prepared and compared with the controls.. The chromosome aberration and micronuclei rates were not significantly different from those of the controls (P > 0.05).. The results were in accordance with our previous reports about the effects of T4 and T7 on the chromosome aberration and micronuclei rates of rat spermatogenic cells. At the antifertility doses, T4 and T7 did not show a mutagenic effect.

Topics: Animals; Bone Marrow Cells; Chromosome Aberrations; Diterpenes; Epoxy Compounds; Infertility, Male; Male; Micronuclei, Chromosome-Defective; Phenanthrenes; Rats; Rats, Sprague-Dawley; Tripterygium; Triterpenes