Role of nitric oxide donor in methotrexate-induced testicular injury via modulation of pro-inflammatory mediators, eNOS and P-glycoprotein

Abstract

Methotrexate (MTX) is a widely used chemotherapeutic agent but its clinical use is challenged with different forms of toxicities including testicular injury. The aim of the current study was to evaluate the potential protective effect of potassium channel opener, nicorandil (NIC) (3 and 10 mg/kg/day) on MTX-induced testicular injury in a rat model. Rats were randomly divided into four groups (nine rats each) and treated for 2 weeks as follows: (I) normal control (CON group) received vehicle, (II) model group (MTX group) given MTX (20 mg/kg) single intraperitoneal ( i.p.) injection dose on 11th day, (III) MTX + NLD group treated with NIC (3 mg/kg/day) orally for 2 weeks and MTX (20 mg/kg) single i.p. dose on 11th day, and (IV) MTX + NHD group treated with NIC (10 mg/kg/day) orally for 2 weeks and MTX (20 mg/kg) single i.p. injection on the 11th day. The testicular injury was assessed biochemically via serum testosterone, total antioxidant capacity, testicular oxidative stress parameters, P-glycoprotein, tumor necrosis factor-alpha, and interleukin-1β. Furthermore, histopathological evaluation, endothelial nitric oxide synthase (eNOS) immunoexpression, and detection of p53 expression level using Western blotting were performed. Results showed that MTX induced testicular injury which was proved by both biochemical and histopathological evaluations. Our results concluded that NIC pretreatment attenuated MTX-induced testicular injury via significantly increased eNOS immunoexpression, antiapoptotic, anti-inflammatory, and antioxidant properties. Interestingly, NIC high dose is more protective than low dose.

The Possible Contribution of P-Glycoprotein in the Protective Effect of Paeonol against Methotrexate-Induced Testicular Injury in Rats

“…Testicular injury is a serious adverse effect of MTX therapy, thus the preservation of germinal cells is an essential target during cancer chemotherapy [ 6 , 7 ]. In line with previous reports [ 22 , 23 ], MTX treatment in the current study was associated with a significant decrease in testicular weight, indicative of testicular atrophy. Pretreatment with paeonol, however, prevented the MTX-induced testicular atrophy, as shown by preservation of testicular weight.…”

“…Moreover, MTX resulted in testicular functional impairment in the form of significantly decreased serum testosterone level and impaired spermatogenesis. These results are supported by the findings of previous studies [ 23 , 24 , 25 ], which demonstrated that a single dose of MTX was associated with testicular structural damage, impaired spermatogenesis, and decreased testosterone. Alternatively, pretreatment with paeonol mitigated the MTX-induced testicular injury, with marked preservation of normal testicular architecture as well as the functional ability of the treated testes, which is in agreement with our recent findings showing protection by paeonol against testicular ischemia-reperfusion injury [ 19 ].…”

“…For example, Norris et al [ 26 ] reported that MTX led to increased expression of P-gp in CEM/MTX R cells. On the other hand, MTX resulted in decreased rat testicular P-gp level [ 23 ]. This probably means that there is a differential response of P-gp to MTX treatment in non-cancerous versus cancerous cells.…”

“…The inflammatory cytokine TNF-α plays a significant role in the pathogenesis of MTX-induced testicular injury [ 22 , 23 , 24 , 25 ]. TNF-α induces oxidative stress, the release of other inflammatory mediators, NO production, and apoptosis [ 39 , 40 , 41 ], leading to impairment of spermatogenesis [ 35 ] and a decrease in testosterone level [ 42 ].…”

Therapeutic Effect of Stem Cells on Male Infertility in a Rat Model: Histological, Molecular, Biochemical, and Functional Study

“…The H&E-stained testicular sections revealed congestion with interstitial oedema, damage and loss of the germinal cells and Sertoli cells, Leydig cell degeneration, decreased mean germinal epithelium thickness, and tubule diameter indicating decreased spermatogenesis and infertility. These histopathological changes come in agreement with previous studies which reported that MTX induces degeneration and atrophy of seminiferous tubules, sloughing of germ cells away from the basal lamina, decrease in quantity and quality of sperms, and severe apoptosis in the germinal epithelium [ 19 , 21 , 22 ]. Damage to the germinal epithelium resulted in a significant decrease in the Johnson score; the same was found by [ 21 , 23 ].…”