基于内质网自噬系统探究泽泻汤对高脂血症急性胰腺炎的防治作用  

Effect of Zexie Decoction on hyperlipidemia acute pancreatitis based on endoplasmic reticulum-autophagy system

作　　者：张荣展 汤甜甜 黄芳 宋婧 周佳 ZHANG Rongzhan;TANG Tiantian;HUANG Fang;SONG Jing;ZHOU Jia(Department of Traditional Chinese Medicine Pharmacology and Traditional Chinese Medicine,College of Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 211198,China;Laboratory Animal Center,Xiamen University,Xiamen 361102,China)

机构地区：[1]中国药科大学中药学院中药药理与中医药学系,江苏南京211198 [2]厦门大学实验动物中心,福建厦门361102

出　　处：《中草药》2024年第11期3705-3715,共11页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81973410)。

摘　　要：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(50μg/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�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(50μg/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/Ⅰand 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 patholog

关 键 词：泽泻汤 高脂血症急性胰腺炎 氧化应激 线粒体功能障碍 内质网应激 自噬 白术内酯Ⅱ 白术内酯Ⅲ 泽泻醇A 泽泻醇B 23-乙酰泽泻醇B 23-乙酰泽泻醇C 

分 类 号：R285.5[医药卫生—中药学]