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Objective: Variants in neuronal NOS (nNOS) gene were associated with atherosclerosis and stroke susceptibility.We aimed to investigate the association between nNOS gene polymorphism and risk of ischemic stroke caused by small-artery occlusion (SAO) and large-artery atherosclerosis (LAA) in a Chinese population.Methods: We conducted a case-control study involving 381 ischemic stroke patients and 366 healthy subjects.Selected SNPs (rs1483757, rs2293050 and rs2139733) were genotyped and assessed, the association with the risk of ischemic stroke was analyzed.Furthermore, gender and etiological subtype stratified analyses were also carried out to evaluate the association between nNOS polymorphisms and risk of ischemic stroke.Results: No significant difference was observed between selected nNOS loci and risk of ischemic stroke in alleles or any genetic models in total study population, males or females, adjusted with age, drinking and smoking status.Rs2293050 and rs2139733 genotypes were associated with total cholesterol (rs2293050, P=0.026;rs2139733, P=0.040) and LDL (rs2293050, P=0.031;rs2139733, P=0.046) in females.A significant difference in allele distribution of rs2293050 (P =0.040) and a marginally significant difference of rs2139733 (P =0.061) in LAA caused ischemic stroke cases and controls were observed in total population.No association between rs1483757 and ischemic stroke was found in this study.Conclusions: T allele of rs2293050 and A allele of rs2139733 in nNOS gene may contribute to increased susceptibility of large-artery atherosclerosis caused ischemic stroke in Han Chinese.