目的 探讨刺五加注射液对大鼠肾缺血再灌注损伤(RIRI)的保护作用及其机制。方法 将48只健康SD大鼠随机分为假手术组、模型组、刺五加注射液(30 mg/kg)组和刺五加注射液(30 mg/kg) +Nrf2抑制剂(鸦胆子苦醇, 2 mg/kg)组,每组12只。除假手术组外,其他组采用夹闭双侧肾动脉45 min的方法复制RIRI大鼠模型。造模前7 d开始,各组分别ip给药(假手术组和模型组给予0.9%氯化钠溶液), 1次/d。肾缺血再灌注6 h后,计算肾脏指数,检测血清肾功能指标,通过苏木精-伊红(HE)染色法进行肾组织病理学检查;分光光度法检测肾组织抗氧化酶活性和丙二醛(MDA)水平, ELISA法检测炎症因子水平, Western blotting法检测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、胞浆核因子-κB p65(NF-κB p65)、胞核NF-κB p65蛋白表达。结果 与模型组比较,刺五加注射液组大鼠肾脏指数降低(P<0.05),血清尿素氮(BUN)、肌酐(Scr)水平降低(P<0.05),肾组织病理学改变明显改善,病理评分降低(P<0.05);肾组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性升高且MDA、肿瘤坏死因子(TNF-α)、白细胞介素(IL-1β)水平降低(P<0.05);肾组织Nrf2、HO-1表达量升高(P<0.05),胞核NF-κB p65表达量和胞核NF-κB p65/胞浆NF-κB p65值降低(P<0.05)。Nrf2抑制剂鸦胆子苦醇能够明显逆转刺五加注射液对Nrf2、HO-1、胞核NF-κB p65表达的调控作用以及对大鼠RIRI的保护作用。结论 刺五加注射液对大鼠RIRI具有保护作用,其机制可能与激活Nrf2/HO-1通路、抑制NF-κB核转位,进而抑制肾组织氧化应激和炎症反应有关。

Objective To investigate the protective effect and mechanism of Ciwujia Injection on renal ischemia-reperfusion injury (RIRI) in rats.Methods 48 healthy SD rats were randomly divided into sham operation group, model group, Ciwujia Injection (30 mg/kg) group, and Ciwujia Injection (30 mg/kg) + Nrf2 inhibitor (brusatol, 2 mg/kg) group, each group had 12 rats. Except for the sham operation group, the rats in other groups were prepared RIRI rat model by clamping bilateral renal arteries for 45 min. And 7 days before modeling, the rats in each group was given intraperitoneal injection (the rats in the sham operation group and the model group were given 0.9% sodium chloride solution), once daily. 6 Hours after renal ischemia-reperfusion, the renal index was calculated, the level of renal function indexes in serum were detected, the renal histopathological was examined by HE staining method. The activity of antioxidant enzyme and the content of MDA were detected by spectrophotometry, the level of inflammatory factors were detected by ELISA method, the protein expression of Nrf2, HO-1, cytoplasmic NF-κB p65, nuclear NF-κB p65 were detected by Western blotting method.Results Compared with the model group, the renal index of the rats in the Ciwujia Injection group was decreased (P < 0.05), the level of blood urea nitrogen (BUN), creatinine (Scr) in serum were decreased (P < 0.05); the renal histopathological changes was significantly improved, and the pathological score was decreased (P < 0.05). The activity of superoxide dismutase (SOD), catalase (CAT) in renal tissue were increased, and the levels of MDA, TNF-α, IL-1β were decreased (P < 0.05). The expression of Nrf2, HO-1 in renal tissue were increased (P < 0.05), and the expression of nuclear NF-κB p65 and the ratio of nuclear NF-κB p65/cytoplasmic NF-κB p65 were decreased (P < 0.05). The Nrf2 inhibitor brusatol could significantly reverse the regulatory effect of Ciwujia Injection on the expression of Nrf2, HO-1, nuclear NF-κB p65 and the protective effect on RIRI in rats (P < 0.05).Conclusion Ciwujia Injection has protective effect on RIRI in rats, and its mechanism may be related to activating the Nrf2/HO-1 pathway and inhibiting NF-κB nuclear translocation, and then inhibiting oxidative stress and inflammation in the renal tissue.

R285.5

河北省医学科学研究课题(20191258)