CAJ | 학술논문

目的研究萝卜硫素(SFN)对大鼠局部脑缺血-再灌注损伤的神经保护机制.方法 24只雄性SD大鼠,随机(随机数字法)分为3组,A组:假手术组(n=8),B组:缺血-再灌注模型组(n=8),C组:萝卜硫素干预组(n=8);A组:假手术后腹腔注射等量磷酸盐缓冲液(PBS),B组:缺血-再灌注术后腹腔注射等量PBS,C组:缺血-再灌注术后腹腔注射萝卜硫5 mg/kg.通过四氮唑红(2,3,5-Triphenyltetrazolium chloride,TTC)染色测定脑梗死体积,苏木精伊红染色法(HE)观察组织学形态改变,TUNEL法检测脑神经元凋亡变化,RT-PCR测定缺血脑组织核转录因子kappa B亚单位p65(NF-κBp65)、诱导型一氧化氮合酶(iNOS) mRNA的表达,免疫蛋白印记法(Western-bloting)检测脑组织iNOS、NF-κBp蛋白表达水平.结果与B组比,C组脑梗死体积降低[(96.34±3.72) vs.(124.65 ±3.85),P＜0.01],且脑神经元凋亡减少,RT-PCR法显示C组脑组织NF-κBpmRNA和iNOS mRNA含量降低[(0.26 ±0.018)vs.(0.43±0.031),P＜0.01].Western-bloting结果显示,C组NF-κB和iNOS含量降低[(0.67 ±0.042) vs.(0.56 ±0.032),P＜0.01].结论 SFN可通过减少脑局灶性缺血-再灌注损伤后缺血脑组织NF-κB和iNOS表达,减少脑梗死体积,抑制神经元细胞凋亡,具有脑神经保护作用.
목적 연구라복류소(SFN)대대서국부뇌결혈-재관주손상적신경보호궤제.방법 24지웅성SD대서,수궤(수궤수자법)분위3조,A조:가수술조(n=8),B조:결혈-재관주모형조(n=8),C조:라복류소간예조(n=8);A조:가수술후복강주사등량린산염완충액(PBS),B조:결혈-재관주술후복강주사등량PBS,C조:결혈-재관주술후복강주사라복류5 mg/kg.통과사담서홍(2,3,5-Triphenyltetrazolium chloride,TTC)염색측정뇌경사체적,소목정이홍염색법(HE)관찰조직학형태개변,TUNEL법검측뇌신경원조망변화,RT-PCR측정결혈뇌조직핵전록인자kappa B아단위p65(NF-κBp65)、유도형일양화담합매(iNOS) mRNA적표체,면역단백인기법(Western-bloting)검측뇌조직iNOS、NF-κBp단백표체수평.결과 여B조비,C조뇌경사체적강저[(96.34±3.72) vs.(124.65 ±3.85),P＜0.01],차뇌신경원조망감소,RT-PCR법현시C조뇌조직NF-κBpmRNA화iNOS mRNA함량강저[(0.26 ±0.018)vs.(0.43±0.031),P＜0.01].Western-bloting결과현시,C조NF-κB화iNOS함량강저[(0.67 ±0.042) vs.(0.56 ±0.032),P＜0.01].결론 SFN가통과감소뇌국조성결혈-재관주손상후결혈뇌조직NF-κB화iNOS표체,감소뇌경사체적,억제신경원세포조망,구유뇌신경보호작용.
Objective To investigate the protective effects of sulforaphen (SFN) on focal cerebral ischemia/reperfusion injuy (IRI) in rats in order to explore the mechanisms.Methods Twenty-four male SD rats were randomly (random number) divided into Sham-operated group (A group,n =8),IRI group (B group,n =12),sulforaphen group (C group,n =8).SD rats were made to be transient focal cerebral IRI models.SFN 5 mg/kg was injected intraperitoneally to rats 15 minutes after IRI in C group,and rats of group A and group B received equal volume PBS instead.Infarct volume was measured by TTC staining and morphologic changes were observed with HE staining.Neuronal cell apoptosis index was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) assay.Rats were sacrificed at 24 h after IRI.The protein levels of NF-κB p65 and iNOS were detected by using western bloting and the mRNA expressions of NF-κB p65 and iNOS were detected by using RT-PCR.Results Compared with the group B,infarct volume was significantly smaller in group C,the number of neuronal cell apoptosis in brain tissue were decreased significantly in group C [(96.34 ±3.72) vs.(124.65 ±3.85),P ＜ 0.01],the levels of NF-κB and iNOS in brain tissue of rats were decreased in the SFN group (P ＜ 0.01).SFN reduced neuronal cell apoptosis,injury,and infarct volume [(0.26 ± 0.018) vs.(0.43 ±0.031),P ＜0.01].The mRNA expression and protein level of NF-κBp65 were decreased in the group C.And the mRNA expression and protein level of induced nitric oxide synthase (iNOS) in IRI affected brain tissue were decreased in the group C [(0.67 ± 0.042) vs.(0.56 ± 0.032),P ＜ 0.01].Conclusions SFN might decrease the neuronal cell apoptosis caused by ischemia/repeffusion injury,and this protective effect is mediated by decreasing the level of NF-κB and iNOS.