Nerve growth factor alleviates arsenic-induced testicular injury by enhancing the function of Sertoli cells.

Sertoli cells (SCs) maintain testicular homeostasis and promote spermatogenesis by forming the blood-testis barrier (BTB) and secreting growth factors. The pro-proliferative and anti-apoptotic effects of nerve growth factor (NGF) on SCs have been proved previously. It is still unclear whether the damage effect of arsenic on testis is related to the inhibition of NGF expression, and whether NGF can mitigate arsenic-induced testicular damage by decreasing the damage of SCs induced by arsenic. Here, the lower expression of NGF in testes of arsenic exposed mice (freely drinking water containing 15 mg/l of NaAsO2) was observed through detection of Western blot and Real-time PCR. Subsequently, hematoxylin and eosin (HE) staining, Evans blue staining and transmission electron microscopy were used to evaluate the pathology, BTB permeability and tight junction integrity in testes of control mice, arsenic exposed mice (freely drinking water containing 15 mg/l of NaAsO2) and arsenic + NGF treated mice (freely drinking water containing 15 mg/l of NaAsO2 + intraperitoneal injection with 30 µg/kg of NGF), respectively. Evidently, spermatogenic tubule epithelial cells in testis of arsenic exposed mice were disordered and the number of cell layers was reduced, accompanied by increased permeability and damaged integrity of the tight junction in BTB, but these changes were less obvious in testes of mice treated with arsenic + NGF. In addition, the sperm count, motility and malformation rate of mice treated with arsenic + NGF were also improved. On the basis of the above experiments, the viability and apoptosis of primary cultured SCs treated with arsenic (10 µM NaAsO2) or arsenic + NGF (10 µM NaAsO2 + 100 ng/mL NGF) were detected by Cell counting kit-8 (CCK8) and transferase-mediated DUTP-biotin nick end labeling (TUNEL) staining, respectively. It is found that NGF ameliorated the decline of growth activity and the increase of apoptosis in arsenic-induced SCs. This remarkable biological effect that NGF inhibited the increase of Bax expression and the decrease of Bcl-2 expression in arsenic-induced SCs was also determined by western blot and Real-time PCR. Moreover, the decrease in transmembrane resistance (TEER) and the expression of tight junction proteins ZO-1 and occludin was mitigated in SCs induced by arsenic due to NGF treatment. In conclusion, the above results confirmed that NGF could ameliorate the injury effects of arsenic on testis, which might be related to the function of NGF to inhibit arsenic-induced SCs injury.

D-Limonene Alleviates Oxidative Stress Injury of the Testis Induced by Arsenic in Rat.

Long-term exposure to arsenic can lead to testicular damage and lower sperm quality in males, which is mediated by increased arsenic-induced oxidative stress and other damage mechanisms. D-Limonene, which is rich in oranges, lemons, oranges, grapes and other natural fruits, can relieve doxorubicin (DOX)-induced kidney injury and CCL4-induced cardiac toxicity by inhibiting oxidative stress and inflammatory response. The antioxidant and anti-inflammatory properties of D-limonene motivate us to further explore whether it can reduce arsenic-induced testicular injury. To verify this scientific hypothesis, testicular pathology, testicular oxidative stress levels and sperm motility were determined after intervention with D-limonene in rats chronically exposed to arsenic. As expected, long-term arsenic exposure caused testicular tissue structure disturbances, increased levels of oxidative stress, and decreased sperm activation, all of which were significantly inhibited due to treatment with D-limonene. In conclusion, our data reveal a previously unproven beneficial effect of D-limonene, namely that D-limonene can inhibit arsenic-induced testicular injury, and also provide theoretical and experimental basis for the application of D-limonene in the treatment of arsenic-induced testicular injury.