目的 研究泽泻汤对高脂血症急性胰腺炎（hyperlipidemia acute pancreatitis，HLAP）的防治作用及机制。方法 雄性C57BL/6J小鼠以高脂饮食喂养，ig泽泻汤干预8周后，ip雨蛙素（50 μg/kg）制备HLAP模型。采用试剂盒检测小鼠血清中三酰甘油（triglyceride，TG）、总胆固醇（total cholesterol，TC）水平及淀粉酶（amylase，AMY）、脂肪酶（lipase，LPS）、总超氧化物歧化酶（total superoxide dismutase，T-SOD）、过氧化氢酶（catalase，CAT）活性；采用油红O染色观察肝脏组织脂质蓄积情况，苏木素-伊红（HE）染色观察肝脏、胰腺组织病理学变化；采用Western blotting检测胰腺组织中血红素加氧酶-1（heme oxygenase-1，HO-1）、核因子E2相关基因（nuclear factor erythroid-2-related factor 2，Nrf2）、免疫球蛋白重链结合蛋白（binding immunoglobulin protein，BIP）、真核翻译起始因子2α（eukaryotic translation initiation factor 2α，eIF2α）、p-eIF2α、Bcl-2相互作用蛋白（Bcl-2 interacting coiled-coil protein-1，Beclin-1）、自噬蛋白5（autophagy-related protein 5，ATG5）、自噬蛋白12（autophagy-related protein 12，ATG12）、微管相关蛋白1轻链3-II/I（microtubule-associated protein light chain 3-II/I，LC3-II/I）及溶酶体相关膜蛋白1（lysosomal associated membrane protein 1，LAMP-1）表达。采用棕榈酸（200 mmol/L）和雨蛙素（10 nmol/L）处理胰腺腺泡AR42J细胞构建HLAP细胞模型，给予泽泻汤干预后，采用试剂盒检测细胞培养上清液中LPS、T-SOD活性；采用DCFH-DA荧光探针、线粒体膜电位（mitochondrial membrane potential，MMP）荧光探针检测细胞内活性氧（reactive oxygen species，ROS）及MMP水平；采用透射电镜观察细胞超微结构及自噬变化；采用Western blotting检测细胞中HO-1、Nrf2、Kelch样环氧氯丙烷相关蛋白1（kelch-like ECH-associated protein 1，Keap1）、BIP、eIF2α、p-eIF2α、Beclin-1、ATG5、LC3-II/I和LAMP-1表达。结果 与模型组比较，泽泻汤能够降低小鼠血清中TC、TG水平及AMY、LPS活性（P＜0.05、0.01、0.001），抑制肝脏组织中的脂滴蓄积，减轻肝脏及胰腺组织病理损伤，同时上调血清及胰腺腺泡细胞中SOD、CAT活性（P＜0.05、0.01），下调ROS水平（P＜0.001），上调胰腺组织及细胞中HO-1、Nrf2、LAMP1表达（P＜0.05、0.01、0.001），下调Keap1、BIP、eIF2α、p-eIF2α、Beclin-1、ATG5、ATG12及LC3-II/I表达（P＜0.05、0.01、0.001）。结论 泽泻汤可以防治HLAP，其作用机制可能与抑制内质网应激诱导的过度自噬、恢复自噬通量有关。

Objective To investigate the preventive effect and mechanism of Zexie Decoction (泽泻汤) on hyperlipidemia acute pancreatitis (HLAP). Methods Male C57BL/6J mice were fed with a high-fat diet and ig Zexie Decoction for eight weeks, HLAP model was prepared by ip cerulein (50 μg/kg). Levels of triglyceride (TG), total cholesterol (TC) and activities of amylase (AMY), lipase (LPS), total superoxide dismutase (T-SOD), catalase (CAT) in the serum of mice were detected by kit; Lipid accumulation in liver tissue was observed by oil red O staining, histopathological changes in liver and pancreatic tissue were observed by HE staining; The protein expressions of heme oxygenase-1 (HO-1), nuclear factor erythroid-2-related factor 2 (Nrf2), kelch-like ECH-associated protein 1 (Keap1), binding immunoglobulin protein (BIP), eukaryotic translation initiation factor 2α (eIF2α), p-eIF2α, Bcl-2 interacting coiled-coil protein 1 (Beclin-1), autophagy-related protein 5 (ATG5), autophagy-related protein 12 (ATG12), microtubule-associated protein light chain 3-II/I (LC3-II/I) and lysosomal associated membrane protein-1 (LAMP-1) in pancreatic tissue were detected by Western blotting. AR42J cells were incubated with palmitic acid (200 mmol/L) and cerulein (10 nmol/L) to establish the HLAP cell model after intervention with Zexie Decoction, activities of LPS and T-SOD in culture supernatant of cells were detected by kit; The DCFH-DA fluorescent probe and mitochondrial membrane potential (MMP) fluorescent probe were used to detect intracellular reactive oxygen species (ROS) and MMP levels; The changes in ultrastructure and autophagy of cells were observed by electron microscopy; The protein expressions of HO-1, NRF2, kelch-like ECH-associated protein 1 (Keap1), BIP, eIF2α, p-eIF2α, Beclin-1, ATG5, LC3-II/I and LAMP-1 in cells were detected by Western blotting. Results Compared with the model group, Zexie Decoction reduced the levels of TC, TG and activities of AMY and LPS in serum of mice (P < 0.05, 0.01, 0.001), inhibited the accumulation of lipid droplets in liver tissues, attenuated the pathological damage of liver and pancreatic tissues, up-regulated the activities of SOD and CAT in serum and pancreatic acinar cells (P < 0.05, 0.01), down-regulated the level of ROS (P < 0.001), up-regulated the expressions of HO-1, Nrf2 and LAMP1 in pancreatic tissues and cells (P < 0.05, 0.01, 0.001), down-regulated the expressions of Keap1, BIP, eIF2α, p-eIF2α, Beclin-1, ATG5, ATG12 and LC3-II/I (P < 0.05, 0.01, 0.001). Conclusion Zexie Decoction can prevent HLAP and its mechanism may be related to the inhibition of endoplasmic reticulum stress-induced excessive autophagy and restoration of autophagic flux.

R285.5

国家自然科学基金面上项目（81973410）