萝卜硫素对肾病综合征大鼠肾脏损伤保护的机制研究

Protective effect of sulforaphane on kidney injury in rats with nephrotic syndrome

目的研究萝卜硫素对肾病综合征大鼠肾脏损伤的保护作用及其对一氧化氮合酶表达的影响。方法采用5 mg/kg阿霉素诱导肾病综合征大鼠模型,将大鼠分为对照组、模型组、萝卜硫素组及阳性药物组,30 mg/kg萝卜硫素和6 mg/kg阳性药物贝那普利分别灌胃治疗大鼠6周,HE染色检测肾脏组织病理学变化,BCA法检测24 h尿蛋白量,ELISA法检测了血浆白蛋白水平,分光光度法测定尿液NO含量;Western blot法检测肾组织内皮型一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)和神经型一氧化氮合酶(nNOS)的表达。结果相对于对照组大鼠,模型组大鼠24 h尿蛋白量明显升高(P<0.05),血浆白蛋白明显降低(P<0.05),肾脏病理损伤严重,尿液中NO含量显著降低(P<0.05),eNOS、iNOS及nNOS酶表达水平亦显著降低(P<0.05);相比于模型组大鼠,萝卜硫素和阳性药物治疗组大鼠24 h尿蛋白量明显降低(P<0.05),血浆白蛋白明显升高(P<0.05),肾脏病理损伤明显改善,尿液中NO含量显著升高(P<0.05),eNOS、iNOS及nNOS酶表达水平亦显著增高(P<0.05)。结论萝卜硫素能够诱导肾病综合征大鼠eNOS、iNOS及nNOS的表达,提升大鼠体内NO水平,降低尿蛋白含量,改善肾脏损伤程度。

Objective To study the effects of sulforaphane on kidney injury and the expression levels of nitric oxide synthase in rats with nephrotic syndrome.Methods Five mg/kg of doxorubicin was used to construct the nephrotic syndrome model in rats.Rats were divided into control group,model group,sulforaphane treatment group and benazepril(positive drug)treatment group.After treatment with 30 mg/kg of sulforaphane and 6 mg/kg of benazepril for 6 weeks,HE staining was used to analyze pathological changes in kidney tissue in rats with nephrotic syndrome.Proteinuria level in 24 hours was measured by BCA assay.ELISA assay was used to detect the level of plasma albumin.NO content was detected by spectrophotometer.Western blot was used to determine the expression levels of endothelial NOS(eNOS),inducible NOS(iNOS)and neural NOS(nNOS).Results Compared with control group,the 24 hours proteinuria level was significantly increased in model group(P<0.05),and the pathological changes of the kidney were serious.The urine NO content of the model group was greatly decreased(P<0.05),the level of plasma albumin was greatly decreased(P<0.05),and the expression levels of eNOS,iNOS and nNOS were also significantly decreased(P<0.05).Compared with model group,the 24 hours proteinuria level was significantly increased in sulforaphane treatment group and benazepril treatment group group(P<0.05),the level of plasma albumin was greatly increased(P<0.05),and the pathological changes of the kidney were significantly improved.The urine NO content of the treatment groups was greatly increased(P<0.05),and the expression levels of eNOS,iNOS and nNOS were also significantly increased(P<0.05).Conclusions Sulforaphane can improve the expressions of eNOS,iNOS and nNOS in nephrotic syndrome rats,increase the content of NO in rats,reduce the content of urinary protein,and improve the pathological process of nephrotic syndrome.