Stroke is the second leading cause of death and permanent disability worldwide. The role of nitric oxide (NO) inhibition in cerebral ischemia/reperfusion (I/R) remains controversial (1). Rho-kinases (ROCK) are serine-threonine kinases that play a vital role in cell migration and survival, interaction between NO and ROCK isn’t clear (2). Nuclear factor kappa β (NF-kβ) is a transcription factor that contributes to infarction in experimental stroke and its inhibition associated with neuroprotective effects (3). This study investigated the potential neuroprotective effect of nonselective nitric oxide synthase inhibitor (L-NAME) in rat’s transient cerebral I/R. 30 adult male Wistar rats (150-250g) were divided into three groups 10 rats in each: first group was sham-operated and served as a control, I/R group of rats infused with 0.9% normal saline intraperitoneally 15 minutes prior to 30 minutes of left common carotid artery occlusion, followed by 24-hour of reperfusion, anesthesia was induced by ether inhalation and maintained by thiopental sodium (2.5mg/kg) (4). A test group infused with L-NAME (15 mg/kg per weight) intraperitoneally 15 minutes prior to same I/R period. Neurobehavioral assessments were evaluated (5), NF-κβ via western blotting and ROCK using enzyme-linked immunosorbent assay were estimated in the left cerebral hemisphere. NO metabolites (nitrite and nitrate) were measured colorimetrically in plasma and brain tissue. Values are means ± S.E.M., compared by ANOVA. The L-NAME group showed a significant improvement in neurological deficit (15.07±0.584) compared to both I/R and control groups (12.798±0.689, 17.50±0.707, respectively P <0.001). In I/R group; NF-κβ was significantly increased (129.2±1.7 RGB unit/mg protein) compared to the control group (53±1.03RGB unit/mg protein, P <0.001) and L-NAME pretreatment resulted in significant decrease in NF-κβ (44.4±1.3RGB unit/mg protein, P <0.001) compared to I/R rats. In I/R group brain tissue levels of ROCK and NO significantly increased (0.022±0.0054 ng/mg protein, 8.88±0.572 nmol/mg protein) compared to the control group (0.013±0.003 ng/mg protein, 3.48±0.228 nmol/mg protein, P <0.001). The L-NAME group showed a significant decrease in tissue level of ROCK and NO (0.005±0.0006 ng/mg protein, 4.47±0.392 nmol/mg protein, P <0.001) compared to the I/R group. Serum level NO was significantly increased in I/R group (42.03±4.558 μmol/L) compared to the control group (17.84±0.701 μmol/L, P <0.001), while L-NAME administration resulted in significant decrease in serum NO (18.44±0.513μmol/L, P <0.001) compared to the I/R group. Thus, L-NAME significantly improved neurological deficit and inflammation evidence by decrease NF-κβ and ROCK in the affected cerebral hemisphere following cerebral ischemic injury.