莱菔硫烷激动Nrf-2抗炎症改善小鼠肾缺血再灌注损伤

Sulforaphane attenuates renal ischemia reperfusion injury in mice by Nrf-2 against inflammation

目的 基于小鼠肾缺血再灌注(ischemia reperfusion injury, IRI)模型探析莱菔硫烷(sulforaphane, SFN)对小鼠肾IRI的作用与机制,为制定肾缺血再灌注的有效干预措施提供实验依据。方法 24只雄性C57小鼠,随机分为 4组:假手术对照组(Ctrl)、肾缺血再灌注组(IRI)、莱菔硫烷干预肾缺血再灌注组(IRI+SFN)、莱菔硫烷单独给药组(SFN)。术前24 h对SFN处理组小鼠尾静脉注射500 μg/kg SFN。以无创血管夹夹闭小鼠双侧肾蒂45 min建立肾IRI模型,假手术对照组除不夹闭双侧肾蒂外其他操作同上。在各组小鼠恢复肾脏血流灌注24 h以后,收集小鼠全血和肾组织标本进行病理检测。ELISA检测小鼠血清中TNF-α、IL-1β和IL-6的分泌水平。Western blot检测小鼠肾组织中Nrf-2、Keap-1、 p-iκBα、iκBα和p-p65的表达。结果 莱菔硫烷可改善缺血再灌注小鼠肾功能,治疗小鼠肾缺血再灌注损伤并减少肾小管上皮细胞凋亡。莱菔硫烷还可降低缺血再灌注损伤小鼠血清中TNF-α、IL-1β和IL-6的水平,增加缺血再灌注损伤小鼠肾组织中Nrf-2的表达,减少缺血再灌注损伤小鼠肾组织中p-iκBα和胞核内p-p65中的表达。结论 莱菔硫烷可以增加Nrf-2表达,抑制磷酸化iκBα的表达,进而抑制NFκB p65的磷酸化转位进入细胞核并阻止其启动下游炎症因子如TNF-α、IL-1β和IL-6的表达,从而抑制缺血再灌注损伤所致的炎症,改善小鼠肾缺血再灌注损伤。

Objective To explore the effect and mechanism of sulforaphane on renal ischemia reperfusion injury (IRI) in mice to provide experimental evidence for effective intervention in renal IRI.Methods Totally 24 male C57 mice were randomly divided into four groups:control group (Ctrl), IRI group, sulforaphane intervention IRI group (IRI + SFN), and sulforaphane alone group (SFN).SFN was injected into the mice at the dosage of 500 μg/kg 24 h before surgery.The renal IRI model was established by using the bilateral renal pedicles of mice with non-invasive vascular clamping for 45 min.After 24 h of renal blood perfusion, we sacrificed the mice and collected the whole blood and kidney tissue samples for pathological and biological examinations.The levels of TNF-α, IL-1β and IL-6 in murine serum were tested by ELISA.The expression levels of Nrf-2, keap-1, p-iκBα, IκBα and NFκB p-p65 in murine kidney were detected by Western blot.Results Sulforaphane could improve renal function in mice with ischemia reperfusion and reduce apoptosis of renal tubular epithelial cells.Sulforaphane could also decrease the levels of TNF-α, IL-1β and IL-6 in murine serum and increase the expression of Nrf-2 in murine kidney tissue after ischemia reperfusion.Moreover, sulforaphane decreased the expression of p-iκBα in total protein and NFκB p-p65 in nucleus protein of renal tissue.Conclusion Sulforaphane could inhibit phosphorylation, thus inhibiting the phosphorylation of NFκB p65 to translocation into the nucleus.Further it prevented the expressions of downstream inflammatory factors such as TNF-α, IL-1β and IL-6.Finally sulforaphane attenuated renal IRI in mice by Nrf-2 against inflammation.