目的 探讨盐酸川芎嗪注射液对大鼠肝脏缺血再灌注损伤及E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)通路的影响。方法 按随机数字表法将100只大鼠分为假手术组、模型组、丹参注射液(2mL/kg)组、盐酸川芎嗪注射液(30 mg/kg)组和盐酸川芎嗪注射液(30 mg/kg)+鸦胆子苦醇(Nrf2抑制剂,2 mg/kg)组。除假手术组外,各组大鼠采用阻断肝门静脉和肝动脉1h的方法制备肝脏缺血再灌注损伤大鼠模型,造模前30 min ip相应药物。造模6h后,采用生化分析法检测大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBIL)水平;苏木精-伊红(HE)染色法检测肝组织病理学改变;比色法检测肝组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,ELISA法检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6含量,Western blotting法检测核因子E2相关因子2/血红素加氧酶1(Nrf2/HO-1)通路相关蛋白表达水平。结果 与模型组比较,丹参注射液组和盐酸川芎嗪注射液组血清ALT、AST、TBIL水平显著降低(P<0.05),肝组织病理学改变明显改善,病理评分显著降低(P<0.05);肝组织SOD、GSH-Px活性显著升高,且MDA、TNF-α、IL-1β、IL-6含量显著降低(P<0.05),Nrf2、HO-1表达量显著升高(P<0.05),胞核核因子-κB(NF-κB)p65表达量及胞核NF-κB p65/胞浆NF-κB p65比值降低(P<0.05)。Nrf2抑制剂鸦胆子苦醇能够明显逆转盐酸川芎嗪注射液对Nrf2/HO-1通路的激活作用以及对大鼠肝脏缺血再灌注损伤的保护作用。结论 盐酸川芎嗪注射液可能通过激活Nrf2/HO-1通路、抑制NF-κB核转位减轻肝脏缺血再灌注损伤大鼠氧化应激和炎症损伤,对肝脏缺血再灌注损伤起到保护作用。

Objective To investigate the effect of Ligustrazine Hydrochloride Injection on hepatic ischemia-reperfusion injury and E2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway in rats. Methods According to the random number table method, 100 rats were equally divided into sham operation group, model group, Salvia Miltiorrhiza Injection (2 mL/kg) group, Ligustrazine Hydrochloride Injection (30 mg/kg) group, and Ligustrazine Hydrochloride Injection (30 mg/kg) + brusatol (Nrf2 inhibitor, 2 mg/kg) group. Except for the sham operation group, rats in each group were treated with blocking the hepatic portal vein and hepatic artery for 1 h to prepare the rat model of hepatic ischemia reperfusion injury, and corresponding drugs were used 30 min before the model. 6 h after hepatic ischemia-reperfusion injury, the level of ALT, AST, TBIL in serum were detected by biochemical analysis method, the pathological changes of hepatic tissue were detected by HE staining method. The activity of SOD, GSH-Px, and the content of MDA in hepatic tissue was detected by colorimetry. The content of TNF-α, IL-1β, IL-6 were detected by ELISA method, the expression of Nrf2/HO-1 pathway related proteins were detected by Western blotting method. Results Compared with the model group, the level of ALT, AST, and TBIL of Salvia Miltiorrhiza Injection group and Ligustrazine Hydrochloride Injection group were decreased (P＜0.05). The hepatic pathological changes was improved, and the pathological score was decreased (P＜0.05). The activity of SOD, GSH-Px in hepatic tissue were increased, and the content of MDA, TNF-α, IL-1β, and IL-6 were decreased (P＜0.05), but the expression of Nrf2, HO-1 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 obviously reverse the activation effect of ligustrazine hydrochloride injection on the Nrf2/HO-1 pathway and the protective effect on hepatic ischemia-reperfusion injury in rats. Conclusion Ligustrazine Hydrochloride Injection can reduce oxidative stress and inflammatory injury in hepatic ischemia-reperfusion injury rats by activating the Nrf2/HO-1 pathway, and inhibiting NF-κB nuclear translocation, which play a protective role against hepatic ischemia-reperfusion injury.

R965

河北省医学科学研究课题计划(20201142)