广藿香醇对脂肪酸诱导原代小鼠肝细胞脂质积蓄的作用及机制研究  

The effect and mechanism of patchouli alcohol on fatty acid-induced lipid accumulation in primary mouse hepatocytes

作　　者：唐东晖 钟映芹 林重 韩亮 钟艳花 TANG Donghui;ZHONG Yingqin;LIN Zhong;HAN Liang;ZHONG Yanhua(Guangzhou Liwan Hospital of Traditional Chinese Medicine,Guangzhou 510140,China;Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广州市荔湾区中医医院针康科,广东广州510140 [2]广东药科大学健康学院,广东广州510006

出　　处：《广东药科大学学报》2022年第1期75-82,共8页Journal of Guangdong Pharmaceutical University

基　　金：广东省自然科学基金面上项目(2021A1515011280);广东省中医院管理局项目(20201275)。

摘　　要：目的研究广藿香醇对油酸(OA)+棕榈酸(PA)诱导小鼠原代肝细胞脂质积蓄的作用及机制。方法将小鼠原代肝细胞(MPH)分为对照组(DMSO)、模型组(OA+PA)、广藿香醇低、中、高剂量组(5、10、20μmol/L)和奥贝胆酸组(OCA,10μmol/L),采用1 mmol/L的OA+PA处理MPH细胞24 h构建脂质积蓄细胞模型。药物处理24 h后,采用双荧光素酶报告基因、免疫荧光、油红O染色、全自动生化仪和实时荧光定量PCR等技术分别检测广藿香醇对FXR信号激动的作用、FXR蛋白表达和活性氧(ROS)的形成、脂滴聚集、细胞内三酰甘油(TG)含量以及脂质合成相关基因的表达。结果广藿香醇可加强MPH细胞FXR的蛋白表达水平,提高BSEP-luc荧光素酶报告基因活性。与对照组比较,模型组中MPH细胞内脂滴聚集和TG水平明显增加,ROS形成明显升高,脂质合成相关基因(FASN、CD36和FABP1)的mRNA表达水平显著升高,而FXR信号基因表达水平(FXR、SHP和BSEP)则显著降低,差异有统计学意义(P<0.01)。与模型组比较,给药组均可明显降低细胞内红色脂滴聚集和细胞内TG水平,降低ROS水平,抑制脂质合成基因FASN、CD36和FABP1的表达,提高FXR、SHP和BSEP的表达水平,差异有统计学意义(P<0.05,P<0.01)。FXR基因敲除能显著减弱广藿香醇上述的改善作用(P>0.05)。结论广藿香醇通过激动FXR信号,降低脂质合成和ROS水平,从而减少MPH细胞中脂质蓄积。Objective To study the effect and mechanism of patchouli alcohol on lipid accumulation in mouse primary hepatocytes(MPH)induced by oleic acid+palmitic acid(OA+PA).Methods MPH were divided into control group(DMSO),model group(OA+PA)and low,medium and high dose groups(5,10,20μmol/L)and obicholic acid group(OCA,10μmol/L).The MPH cells were treated with 1 mmol/L OA+PA for 24 hours to induce the lipid accumulation cell model.Then the effect of patchouli alcohol on FXR signaling was determined by gene activity analysis using double luciferase reporter.The expression of FXR protein and the formation of reactive oxygen species(ROS)in MPH cells were measured by immunofluorescence.The aggregation of lipid droplets was observed by oil red O staining.The content of intracellular triglyceride(TG)was detected by automatic biochemical instrument.The expression of genes related to lipid synthesis was detected by quantitative real-time PCR(qRT-PCR).Results Patchouli alcohol could promote the protein expression level of FXR in MPH cells,increase the BSEP-luc luciferase reporter gene activity,and induce the mRNA expression levels of FXR,SHP and BSEP in MPH cells.The control group,the oil red O staining experiment in the model group showed that the aggregation of lipid droplets and TG level in MPH cells of model group increased significantly,and also the formation of ROS and the mRNA expression levels of lipid synthesis related genes(FASN,CD36 and FABP1)increased significantly,while the expression levels of FXR signal genes(FXR,SHP and BSEP)decreased significantly(P<0.01).Compared with the model group,patchouli alcohol could significantly reduce the aggregation of intracellular lipid droplets and the level of intracellular TG and ROS inhibit the expression of lipid synthesis genes including FASN,CD36 and FABP1,and increase that of FXR,SHP and BSEP(P<0.05,P<0.01).In FXR knockout MPH cells,compared with the model group,the reduction of patchouli alcohol on the aggregation of intracellular lipid droplets and the reduction of intracel

关 键 词：广藿香醇 FXR 脂质积蓄 氧化应激 脂质合成 

分 类 号：R285.5[医药卫生—中药学]