4-羟基苯并噁唑-2-酮通过脂质代谢和内质网应激通路治疗酒精性脂肪肝大鼠的作用机制研究  

Mechanism of Action of 4-Hydroxy-2(3H)-Benzoxazolone Through Lipid Metabolism and Endoplasmic Reticulum Stress Pathway in the Treatment of Alcoholic Fatty Liver in Rats

作　　者：陆俊霏 刘妍颖 幸尚平 方斌 覃思 张华[2] 陆婷婷 朱丹 林军[1] LU Junfei;LIU Yanying;XING Shangping;FANG Bin;QIN Si;ZHANG Hua;LU Tingting;ZHU Dan;LIN Jun(Pharmaceutical College,Guangxi Medical University,Nanning 530021 Guangxi,China;Department of Pharmacy,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021 Guangxi,China)

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西医科大学第一附属医院药学部,广西南宁530021

出　　处：《中药新药与临床药理》2023年第5期581-590,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81660106,82160734);广西自然科学基金项目(2020GXNSFAA297114);全国大学生创新创业训练计划项目(202010598022)。

摘　　要：目的探讨4-羟基苯并噁唑-2-酮[4-hydroxy-2(3H)-benzoxazolone,HBOA]对酒精性脂肪肝(AFLD)大鼠脂质代谢和内质网应激(ERS)的影响。方法实验前应用AutoDock Tools软件将4-羟基苯并噁唑-2-酮分别与脂质代谢及ERS相关蛋白进行分子对接,预测结合能力。动物实验先将雄性SD大鼠随机分为正常组和模型复制组,参考酒精递增灌胃法建立酒精性脂肪肝模型,模型复制成功的大鼠随机分为模型组、水飞蓟宾组(26.25 mg·kg^(-1))及4-羟基苯并噁唑-2-酮高、中、低剂量组(100、50、25 mg·kg^(-1))。观察大鼠状态;记录体质量、肝质量,并计算肝指数;油红O染色观察肝组织脂肪变性情况;采用试剂盒法或全自动生化分析仪检测肝组织中谷丙转氨酶(GPT)、谷草转氨酶(GOT)和血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平;Western Blot法检测肝组织脂质代谢及ERS相关蛋白固醇调节元件结合蛋白1c(SREBP-1c)、乙酰辅酶A羧化酶1(ACC1)和葡萄糖调节蛋白78(GRP78)、蛋白激酶样ER激酶(PERK)及其磷酸化(p-PERK)、肌醇蛋白酶1(IRE1)及其磷酸化(p-IRE1)、活化转录因子6(ATF6)的表达水平;RT-qPCR法检测肝组织SREBP-1c、GRP78 mRNA表达。结果4-羟基苯并噁唑-2-酮与脂质代谢及ERS相关蛋白之间结合能均小于0 kcal·mol^(-1)。与正常组比较,模型组大鼠肝指数明显增高(P<0.01),镜下可见肝组织大面积红染且细胞内有大量红棕色脂滴,肝组织中GPT、GOT和血清中TC、TG、LDL-C水平明显增高(P<0.01),HDL-C水平明显降低(P<0.01),肝组织中脂质代谢相关蛋白SREBP-1c、ACC1和ERS相关蛋白GRP78、ATF6蛋白表达及PERK、IRE1磷酸化水平明显增高(P<0.01),肝组织中SREBP-1c、GRP78 mRNA相对表达量明显增高(P<0.01)。与模型组比较,4-羟基苯并噁唑-2-酮各组大鼠肝指数(低剂量组除外)明显降低(P<0.01),镜下未见肝组织大面积红染,细胞内脂滴聚集Objective To investigate the effects of 4-hydroxy-2(3H)-benzoxazolone(HBOA)on lipid metabolism and endoplasmic reticulum stress(ERS)in rats with alcoholic fatty liver disease(AFLD).Methods Binding energies of HBOA with lipid metabolism-and ERS-related proteins were predicted using AutoDock Tools software prior to the experiment.In animal experiments,male SD rats were randomly divided into normal group and replicated model group,the model of alcoholic fatty liver was established by alcohol gavage.Rats with successful model-building were then randomly divided into model group,Silibinin group(26.25 mg·kg^(-1)),HBOA high-,medium-,and low-dose group(100,50,25 mg·kg^(-1)).The state of rats was observed,the body mass and liver mass were recorded to calculate liver index.Fatty degeneration of liver tissue was observed by oil red O staining.The measurement of glutamic pyruvic transaminase(GPT)and glutamic oxaloacetic transaminase(GOT)in liver tissue,total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C)and highdensity lipoprotein cholesterol(HDL-C)in serum was performed by kit method or automatic biochemical analyzer.The expressions of lipid metabolism and ERS-related proteins,including sterol regulatory element binding protein 1c(SREBP-1c),acetyl coenzyme A carboxylase 1(ACC1)and glucose regulatory protein 78(GRP78),PKR-like endoplasmic reticulum kinase(PERK)and phosphor-PERK(p-PERK),inositol-requiring enzyme 1(IRE1)and phosphor-IRE1(p-IRE1),and activating transcription factor 6(ATF6)in liver tissue were detected by Western Blot.The mRNA expressions of SREBP-1c and GRP78 in liver tissue was detected by RT-qPCR.Results The binding energies between HBOA and lipid metabolism-or ERS-related proteins were less than 0 kcal·mol^(-1).Compared with the normal group,the liver index was significantly increased in the model group(P<0.01),a large area of red staining in liver tissue and a large number of reddish-brown lipid droplets in the cells were seen under the microscope,the levels of GPT,GOT and s

关 键 词：酒精性脂肪肝 4-羟基苯并噁唑-2-酮 脂质代谢 内质网应激 分子对接 大鼠 

分 类 号：R285.5[医药卫生—中药学]