Wu Xuemei,Wu Jingxian,Yu Shanshan,Zhao Jing,Zhao Yong.Sulforaphane protects neurons against injury induced by oxygen-glucose deprivation/reoxygenation[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1907-1911.
Wu Xuemei,Wu Jingxian,Yu Shanshan,Zhao Jing,Zhao Yong.Sulforaphane protects neurons against injury induced by oxygen-glucose deprivation/reoxygenation[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1907-1911.DOI:
Sulforaphane protects neurons against injury induced by oxygen-glucose deprivation/reoxygenation
目的 探讨莱菔硫烷(SFN)对体外培养皮质神经元氧糖剥夺(OGD)/复氧(R)损伤的保护作用。 方法 出生0~1 d的SD乳大鼠皮质神经元原代培养5~6 d后,随机分为正常对照组、OGD组及SFN组。建立皮质神经元的OGD模型,1 h后复氧,模拟再灌注模型,OGD/R期间给予0.1、0.5、1、2.5 μmol/L SFN,复氧24 h后采用四甲基偶氮唑盐(MTT)微量酶反应比色法测定细胞存活率,测定乳酸脱氢酶(LDH)的释放量来衡量细胞损伤程度,免疫荧光和Western blot观察神经元OGD/R后8-羟基脱氧鸟苷(8-OHDG)和NADPH醌氧化还原酶(NQO1)的表达。 结果 MTT检测示神经元的细胞存活率1 μmol/L SFN组显著高于OGD组[(54.56±8.75)% vs (37.51±9.39)%,P0.01]。SFN组LDH的释放量显著低于OGD组[(330.91±152.33) vs (769.87±275.63)mmol/L,P0.01]。免疫荧光检测示SFN组8-OHDG的阳性表达率显著低于OGD组[(42.10±8.53)% vs (72.50±10.85)%,P0.01],而SFN组NQO1的阳性表达率显著高于OGD组[(90.47±5.46)% vs (70.90±7.92)%,P0.01]。Western blot检测示SFN组NQO1的相对灰度值显著高于OGD组[(0.760±0.063) vs (0.497±0.056),P0.01]。 结论 SFN可减轻OGD/R引起的皮质神经元的损伤。降低8-OHDG的表达和增加NQO1的表达是其可能机制之一。
Abstract
Objective To study the protective effect of sulforaphane (SFN) on in vitro-cultured cortical neuron injury induced by oxygen-glucose deprivation/reoxygenation (OGD/R). Methods Cortical neurons of neonatal SD rats after 5-6 days’ primary culture were randomly divided into control group
OGD group and SFN group. An OGD model of cortical neurons was established. One hour after R
a mimic model of reperfusion was established. During OGD/R
cortical neurons were treated with 0.1 mol/L,0.5 μmol/L,1.0 μmol/L, and 2.5μmol/L SFN
respectively. Twenty-four hours after R
survival rate of cortical neurons was detected by MTT assay
LDH release was measured to determine cortical neuron injury
and expressions of 8-OHDG and NQO1 in cortical neurons were detected by immunofluorescence staining and Western blotting
respectively. Results MTT assay showed that the survival rate of cortical neurons was significantly higher while the LDH release was significantly lower in 1 mol/L SFN group than in OGD group (54.56%±8.75% vs 37.51%±9.39%
330.91±152.33 mmol/L vs 769.87±275.63 mmol/L
P0.01). Immunofluorescence staining displayed that the positive expression rate of 8-OHDG was significantly lower while that of NQO1 was significantly higher in SFN group than in OGD group (42.10%±8.53% vs 72.50%±10.85%
90.47%±5.46% vs 70.90%±7.92%
P0.01). Western blotting showed that the relative grey value of NQO1 was significantly higher in SFN group than in OGD group (0.760±0.063 vs 0.497±0.056
P0.01). Conclusion SFN can reduce cortical neuron injury induced by OGD/R by decreasing the expression of 8-OHDG and increasing the expression of NQO1
which may be one of its protective mechanisms underlying cortical neuron injury.