目的 探討鹽酸川芎嗪注射液對大鼠肝臟缺血再灌注損傷及E2相關(guān)因子2(Nrf2)/血紅素加氧酶1(HO-1)通路的影響。方法 按隨機(jī)數(shù)字表法將100只大鼠分為假手術(shù)組、模型組、丹參注射液(2mL/kg)組、鹽酸川芎嗪注射液(30 mg/kg)組和鹽酸川芎嗪注射液(30 mg/kg)+鴉膽子苦醇(Nrf2抑制劑,2 mg/kg)組。除假手術(shù)組外,各組大鼠采用阻斷肝門靜脈和肝動脈1h的方法制備肝臟缺血再灌注損傷大鼠模型,造模前30 min ip相應(yīng)藥物。造模6h后,采用生化分析法檢測大鼠血清丙氨酸氨基轉(zhuǎn)移酶(ALT)、天冬氨酸氨基轉(zhuǎn)移酶(AST)、總膽紅素(TBIL)水平;蘇木精-伊紅(HE)染色法檢測肝組織病理學(xué)改變;比色法檢測肝組織超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,ELISA法檢測腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素(IL)-1β、IL-6含量,Western blotting法檢測核因子E2相關(guān)因子2/血紅素加氧酶1(Nrf2/HO-1)通路相關(guān)蛋白表達(dá)水平。結(jié)果 與模型組比較,丹參注射液組和鹽酸川芎嗪注射液組血清ALT、AST、TBIL水平顯著降低(P<0.05),肝組織病理學(xué)改變明顯改善,病理評分顯著降低(P<0.05);肝組織SOD、GSH-Px活性顯著升高,且MDA、TNF-α、IL-1β、IL-6含量顯著降低(P<0.05),Nrf2、HO-1表達(dá)量顯著升高(P<0.05),胞核核因子-κB(NF-κB)p65表達(dá)量及胞核NF-κB p65/胞漿NF-κB p65比值降低(P<0.05)。Nrf2抑制劑鴉膽子苦醇能夠明顯逆轉(zhuǎn)鹽酸川芎嗪注射液對Nrf2/HO-1通路的激活作用以及對大鼠肝臟缺血再灌注損傷的保護(hù)作用。結(jié)論 鹽酸川芎嗪注射液可能通過激活Nrf2/HO-1通路、抑制NF-κB核轉(zhuǎn)位減輕肝臟缺血再灌注損傷大鼠氧化應(yīng)激和炎癥損傷,對肝臟缺血再灌注損傷起到保護(hù)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

河北省醫(yī)學(xué)科學(xué)研究課題計(jì)劃(20201142)