目的 探討川芎嗪對(duì)脂多糖（LPS）/D-氨基半乳糖（D-GalN）誘導(dǎo)的急性肝衰竭小鼠的作用及其機(jī)制。方法 將C57BL/6J小鼠隨機(jī)分對(duì)照組、模型組、川芎嗪組、川芎嗪聯(lián)合SIRT1抑制劑（EX527）組，每組10只。通過ip LPS/D-GalN構(gòu)建急性肝衰竭小鼠模型，檢測(cè)血清中天冬氨酸氨基轉(zhuǎn)移酶（AST）、丙氨酸氨基轉(zhuǎn)移酶（ALT）的水平；蘇木精–伊紅（HE）染色檢測(cè)肝組織病理學(xué)變化；實(shí)時(shí)熒光定量PCR（RT-qPCR）檢測(cè)肝組織中腫瘤壞死因子‐α（TNF‐α）、白細(xì)胞介素-6（IL-6）、IL-1β和環(huán)氧化酶-2（COX-2）mRNA水平；檢測(cè)肝組織中氧化應(yīng)激因子谷胱甘肽（GSH）、丙二醛（MDA）和超氧化物歧化酶（SOD）含量；Western blotting檢測(cè)肝組織中沉默調(diào)節(jié)蛋白1（SIRT1）、磷酸化單磷酸腺苷活化蛋白激酶α（p-AMPK）α/AMPKα蛋白和自噬相關(guān)蛋白人微管相關(guān)蛋白1輕鏈3（LC3）-II/LC3-I、p62的水平。結(jié)果 與模型組比較，川芎嗪組小鼠血清ALT和AST水平均顯著降低，肝細(xì)胞壞死區(qū)域以及炎癥細(xì)胞浸潤明顯減少，肝組織中MDA、TNF-α、IL-6、IL-1β、COX-2 mRNA和p62蛋白水平均顯著降低，SOD、GSH、SIRT1、p-AMPKα/AMPKα、LC3-II/LC3-I蛋白水平顯著升高（P<0.05）。EX527處理后顯著逆轉(zhuǎn)了川芎嗪對(duì)急性肝衰竭小鼠的作用。結(jié)論 川芎嗪通過抑制LPS/D-GalN誘導(dǎo)的急性肝衰竭小鼠炎癥反應(yīng)發(fā)揮肝臟保護(hù)作用，其機(jī)制可能與AMPK/SIRT1信號(hào)通路和自噬有關(guān)。

Objective To investigate the effect of ligustrazine on lipopolysaccharide (LPS)/D-galactose (D-GalN) - induced acute liver failure in mice, and its mechanism. Methods C57BL/6J mice were randomly divided into control group, model group, ligustrazine group, and ligustrazine + SIRT1 inhibitor (EX527) group, each group had 10 mice. The mice model of acute liver failure was established by ip LPS/D-GalN, and the serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected in each group. Hematoxylin and eosin (HE) staining was used to detect the pathological changes of liver tissue in each group. mRNA levels of tumor necrosis factor-α (TNF‐α), interleukin (IL)-6, IL-1β, and cyclooxygenase-2 (COX-2) in liver tissue of each group were detected by real-time fluorescent quantitative PCR (RT-qPCR). Contents of oxidative stress factors glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) in liver tissue were detected. Western blotting was used to detect the levels of silencing regulatory protein 1 (SIRT1), phosphorylated adenosine monophosphate activated protein kinase (p-AMPK) α/AMPKα protein and autophagy-related proteins human microtubule-associated protein 1 light chain 3 (LC3) -II/LC3-I and p62 in liver tissue of mice in each group. Results Compared with the model group, the serum levels of ALT and AST in ligustrazine group were significantly decreased, the necrosis area of hepatocytes and the infiltration of inflammatory cells were significantly decreased, and the mRNA and protein levels of MDA, TNF-α, IL-6, IL-1β, and COX-2 in liver tissue were significantly decreased. Protein levels of SOD, GSH, SIRT1, P-AMPKα /AMPKα, and LC3-II/LC3-I were significantly increased (P < 0.05). EX527 treatment significantly reversed the effect of ligustrazine on acute liver failure mice. Conclusion Ligustrazine exerts hepatic protection by inhibiting the inflammatory response in acute liver failure mice induced by LPS/D-GalN, and its mechanism may be related to AMPK/SIRT1 signaling pathway and autophagy.

R965

新疆維吾爾自治區(qū)自然科學(xué)基金項(xiàng)目（2021D01C082）