目的 探討刺五加注射液對大鼠腎缺血再灌注損傷(RIRI)的保護作用及其機制。方法 將48只健康SD大鼠隨機分為假手術(shù)組、模型組、刺五加注射液(30 mg/kg)組和刺五加注射液(30 mg/kg) +Nrf2抑制劑(鴉膽子苦醇, 2 mg/kg)組,每組12只。除假手術(shù)組外,其他組采用夾閉雙側(cè)腎動脈45 min的方法復制RIRI大鼠模型。造模前7 d開始,各組分別ip給藥(假手術(shù)組和模型組給予0.9%氯化鈉溶液), 1次/d。腎缺血再灌注6 h后,計算腎臟指數(shù),檢測血清腎功能指標,通過蘇木精-伊紅(HE)染色法進行腎組織病理學檢查;分光光度法檢測腎組織抗氧化酶活性和丙二醛(MDA)水平, ELISA法檢測炎癥因子水平, Western blotting法檢測核因子E2相關(guān)因子2(Nrf2)、血紅素加氧酶1(HO-1)、胞漿核因子-κB p65(NF-κB p65)、胞核NF-κB p65蛋白表達。結(jié)果 與模型組比較,刺五加注射液組大鼠腎臟指數(shù)降低(P<0.05),血清尿素氮(BUN)、肌酐(Scr)水平降低(P<0.05),腎組織病理學改變明顯改善,病理評分降低(P<0.05);腎組織超氧化物歧化酶(SOD)、過氧化氫酶(CAT)活性升高且MDA、腫瘤壞死因子(TNF-α)、白細胞介素(IL-1β)水平降低(P<0.05);腎組織Nrf2、HO-1表達量升高(P<0.05),胞核NF-κB p65表達量和胞核NF-κB p65/胞漿NF-κB p65值降低(P<0.05)。Nrf2抑制劑鴉膽子苦醇能夠明顯逆轉(zhuǎn)刺五加注射液對Nrf2、HO-1、胞核NF-κB p65表達的調(diào)控作用以及對大鼠RIRI的保護作用。結(jié)論 刺五加注射液對大鼠RIRI具有保護作用,其機制可能與激活Nrf2/HO-1通路、抑制NF-κB核轉(zhuǎn)位,進而抑制腎組織氧化應激和炎癥反應有關(guān)。

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

河北省醫(yī)學科學研究課題(20191258)