N-acetylcysteine effectively mitigates cadmium-induced oxidative damage and cell death in Leydig cells in vitro.

CONTEXT: Cadmium (Cd) is known to cause severe damage to various organs including lung, liver, kidney, brain and reproductive system. Several studies have reported the induction of oxidative stress pathways following Cd exposure. OBJECTIVE: Since oxidative stress is also deemed responsible for inducing male infertility, a growing worldwide concern, we tried to understand whether the antioxidant N-acetylcysteine (NAC) can be a potential therapeutic agent to counter Cd toxicity using primary Leydig cells. MATERIALS AND METHODS: This study highlights the initial cellular alterations which culminate in cell death induction. Primary Leydig cells were isolated from 28-day-old male Wistar rats, exposed to various concentrations of Cd in vitro and biochemical and cell death parameters were evaluated to understand the effect of Cd. NAC pre-treatment was done to understand its protective efficacy. RESULTS: Following Cd exposure to Leydig cells in vitro, we found simultaneous intracellular calcium (Ca(2+)) increase and reduction in mitochondrial membrane polarization at 30 min, followed by significant induction of reactive oxygen species and MAPK-extracellular-regulated kinases with concurrent glutathione depletion at 1 h, and significant cell death (both necrotic and apoptotic) at 6 and 18 h, respectively. Pre-treatment with NAC abrogated all these toxic manifestations and showed significantly reduced cell death. NAC also rescued the expression of 3-ßHSD, a major steroidogenic protein. DISCUSSION AND CONCLUSION: Taken together, these data illustrated that NAC can be used as a potential protective agent against Cd-induced testicular toxicity, especially with regards to oxidative stress-induced Leydig cell toxicity.

Consequences of tributyltin chloride induced stress in Leydig cells: an ex-vivo approach.

Tributyltin (TBT), a member of the organotin family, is a known endocrine disruptor. It persists long in the environment and is widely used in various industrial applications. This study was planned to understand its toxic influence on Leydig cells isolated from 28 day old wistar rats. In-vitro exposure to TBT-Chloride (TBTC) (300-3000 nM) reduced cell viability (DNA fragmentation, nuclear condensation and MTT assay) and affected testosterone production. TBTC induced both apoptotic and necrotic cell death (AnnexinV/PI binding assay). Involvement of calcium (Ca(2+)), redox imbalance (ROS, GSH and TBARS) and mitochondria in TBTC toxicity was evaluated by using Ca(2+) inhibitors (BAPTA-AM, EGTA, Ruthenium Red), free radical scavengers (NAC, C-Phycocyanin) and mitochondrial permeability transition pore inhibitor (Cyclosporine A). Protein expression analysis of phosphorylated MAPKinases (ERK1/2, JNK1/2, & p38), steroidogenic proteins (3ß-HSD, StAR & TSPO) and apoptotic proteins (Bax, Bcl2) illustrates the cytotoxic and anti-steroidogenic activity of TBTC.

Interplay of early biochemical manifestations by cadmium insult in sertoli-germ coculture: an in vitro study.

Cadmium is a common environmental and occupational hazard and its adverse effect on reproductive organ has been well documented. The present study is planned to delineate the mechanism of Cd toxicity in rat testes. Our study shows that Cd causes apoptosis in sertoli-germ cells which is governed by oxidative stress. We assayed ROS, GSH and MMP to ensure the role of oxidative stress, further confirmed it by thiol modulators. The initial biochemical response shown in sertoli-germ cells was a significant rise in intracellular calcium followed by a drastic fall in MMP and then ROS generation. The downstream events included cytochrome c release leading to caspase-3 activation and culminating in cell death via apoptosis. Furthermore Cd disrupted the spermatogenic pathway as evident by suppression in tesmin and LDH-X levels.