摘要
目的探究石斛碱(DNE)通过PPARα调节非酒精性脂肪性肝病(NAFLD)小鼠肝脏脂肪酸代谢的作用及机制。方法将SPF级雄性C57BL/6小鼠随机分为正常饮食组(n=6)、高脂饮食组(n=18),分别给予普通维持饲料和高脂饲料14周,建立NAFLD模型。将高脂饮食组随机平均分为模型组、石斛碱组15 mg/(kg.d)和非诺贝特组27 mg/(kg.d),灌胃给药17周后,收集肝脏样本。采用UPLC-MS/MS、RT-qPCR、Western blotting等技术,分析DNE对小鼠肝脏脂肪酸代谢轮廓及相关靶基因、靶蛋白表达的影响。结果对肝脏中43种脂肪酸进行了定性、定量分析。OPLS-DA结果显示,DNE给药后肝脏脂肪酸的整体轮廓明显区别于模型小鼠,其中含量显著下降的12种脂肪酸均为长链脂肪酸。同时,DNE可显著升高NAFLD小鼠肝脏中过氧化物酶体增殖物激活受体α(PPARα)总蛋白以及核蛋白表达,显著上调与肝脏长链脂肪酸代谢相关的PPARα靶基因Acsl1、Acsl4、Abcd2、Cpt1a、Eci1、Ehhadh的mRNA水平,以及ACOX1、CPT2、HMGCS2、LPL的蛋白表达水平。结论DNE通过核受体PPARα上调肝脏长链脂肪酸代谢相关靶基因的表达,进而影响脂肪酸代谢,改善NAFLD。
Objective To investigate the effects of dendrobine(DNE)on regulating fatty acid metabolism in non-alcoholic fatty liver disease(NAFLD)via PPARαsignaling.Methods SPF-grade male C57BL/6 mice were randomly divided into normal diet group(n=6)and high-fat diet group(n=18),and given normal diet and high-fat diet respectively for 14 weeks to establish NAFLD model.Then the high-fat diet group was randomly and equally divided into model group,DNE group15 mg/(kg.d)and fenofibrate group 27 mg/(kg.d),and administered by gavage for 17 weeks.UPLC-MS/MS,RT-qPCR and western blotting were used to analyze the effects of DNE on liver fatty acid metabolomics and the expression of related target genes and proteins.Results Forty-three fatty acids were analyzed qualitatively and quantitatively.OPLS-DA analysis showed that DNE was able to recover the overall profile of hepatic fatty acid metabolism in NAFLD mice to that of normal liver.The 12 fatty acids whose content was significantly reduced by DNE were all long-chain fatty acids.DNE increased the levels of PPARαtotal protein and nuclear protein in the liver of NAFLD mice.DNE up-regulated the mRNA expressions of Acsl1,Acsl4,Abcd2,Cpt1a,Eci1,and Ehhadh,which are PPARαtarget genes associated with hepatic long-chain fatty acid metabolism.Meanwhile,DNE up-regulated the protein levels of ACOX1,CPT2,HMGCS2 and LPL in mouse liver.Conclusion DNE could ameliorate NAFLD by affecting fatty acid metabolism via the nuclear receptor PPARαand its target genes related to long-chain fatty acid metabolism.
作者
房姝含
许彦哲
罗益
高艺宸
何在霜
李佳颖
陈直
鲁艳柳
Fang Shuhan;Xu Yanzhe;Luo Yi;Gao Yichen;He Zaishuang;Li Jiaying;Chen Zhi;Lu Yanliu(Key Laboratory of Basic Pharmacology of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563006,China;Key Technology Engineering Research Center for the Development of Dendrobium Nobile Industry in Guizhou Colleges and Universities,Zunyi Guizhou 563006,China)
出处
《遵义医科大学学报》
2025年第8期775-782,共8页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:82160812)
国家级大学生创新创业训练计划资助项目(NO:202310661141)
贵州省教育厅项目[NO:黔科技(2022)048]。
作者简介
通信作者:鲁艳柳,女,博士,教授,研究方向:中药药理学,E-mail:yanliu.lu@foxmail.com。
