桑辛素通过调节胆固醇代谢改善油酸诱导的HepG2细胞脂质蓄积  

Lipopexia of Oleic Acid-Induced HepG2 Cells Improved with Morusin by Regulating Cholesterol Metabolism

作　　者：李艳[1,2] 李子雯 李利民 胡胜全[1,2] 吴正治 LI Yan;LI Ziwen;LI Limin;HU Shengquan;WU Zhengzhi(Shenzhen Institute of Translation Medicine,The First Affiliated Hospital of Shenzhen University,Shenzhen 518035,Guangdong,China;Shenzhen Municipal Institute of Geriatric,Shenzhen 518020,Guangdong,China)

机构地区：[1]深圳大学第一附属医院深圳市转化医学研究院,广东深圳518035 [2]深圳市老年医学研究所,广东深圳518020

出　　处：《辽宁中医杂志》2024年第1期140-144,I0004,I0005,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：深圳市科创委基础研究面上项目(JCYJ20220530150408018);国家自然科学基金项目(81803867,81574038);国家科技部重大新药创制项目(2017ZX09301001);深圳市基础研究学科布局项目(JCYJ20220818101806014);广东省自然科学基金项目(2020A1515011427,2020A1515010758);广东省中医药局科研项目(20201319);广东省中医药强省科技专项重点项目(20215002);广东省科技计划项目(2018A0303130078);深圳市医疗卫生三名工程项目(SZSM201612049);深圳市学科布局项目(JCYJ20170412161254416,JCYJ20180508152437368);国家高技术研究发展技术项目(863项目,2014AA020907)。

摘　　要：目的观察桑辛素对油酸(oleic acid,OA)诱导的HepG2细胞脂质蓄积的改善作用,并探讨其可能机制。方法以300μM OA处理HepG2细胞24 h诱导建立脂质蓄积模型,2.5、5、10μM的桑辛素和OA共同作用于细胞24 h,采用CCK-8法测定细胞存活率,油红O染色观察细胞内脂质含量情况,DiI-LDL检测细胞脂质摄取能力,蛋白免疫印迹法(Western blot)测定桑辛素对胆固醇代谢关键基因过氧化物酶体增殖物激活受体a(PPARα)和胆固醇7α-羟化酶(CYP7A1)蛋白表达的情况,并将桑辛素与低密度脂蛋白受体(LDLR)和PPARα通过AutoDock vina软件进行分子对接验证。结果OA刺激HepG2细胞导致细胞内脂滴颗粒明显增加,脂质含量明显升高(P<0.01)。与模型组相比,桑辛素可明显改善模型组肝细胞脂质蓄积,并降低脂质含量(P<0.05)。桑辛素可抑制HepG2细胞对DiI-LDL的摄取,上调细胞中PPARα和CYP7A1的蛋白表达水平(P<0.05)。分子对接显示桑辛素与LDLR和PPARα受体表现出良好的对接活性。结论桑辛素对HepG2细胞脂质蓄积模型有明显的改善作用,其机制可能与调节PPARα通路有关。Objective To observe the effect of morusin on lipid accumulation in HepG2 cells induced by oleic acid(OA)and explore its possible mechanism.Methods HepG2 cells were treated with 300μM OA for 24 h to induce lipid accumulation model.The cells were treated with 2.5,5 and 10μM of morusin and OA for 24 h.The cell survival rate was determined by CCK-8 method,the lipid content in cells was observed by oil red O staining,and the lipid uptake capacity of cells was detected by DiI-LDL.Western blot was used to determine the expressions of peroxisome proliferator-activated receptor A(PPARα)and cholesterol 7α-hydroxylase(CYP7A1)protein,which were key genes of cholesterol metabolism.AutoDock Vina software was used to verify the molecular docking between the low density lipoprotein receptor(LDLR)and PPARα.Results The lipid droplets and lipid content in HepG2 cells were significantly increased by OA stimulation(P<0.01).Compared with model group,morusin could significantly improve lipid accumulation and decrease lipid content in model group(P<0.05).It could inhibit DiI-LDL uptake by HepG2 cells and up-regulate the protein expression levels of PPARαand CYP7A1(P<0.05).Molecular docking showed that mulberry showed good docking activity with LDLR and PPARαreceptors.Conclusion Morusin can significantly improve the lipid accumulation model of HepG2 cells and the mechanism may be related to the pathway of PPARα.

关 键 词：桑辛素 HEPG2细胞 油酸 PPARΑ 脂质蓄积 

分 类 号：R285[医药卫生—中药学]