摘要
目的基于前蛋白转化酶枯草溶菌素9(proprotein convertase subtilisin/kexin type 9,PCSK9)探讨补骨脂素对高脂血症的影响及作用机制。方法首先使用分子对接法对补骨脂素与PCSK9蛋白结合能力进行预测。随后分别采用高脂饮食诱导的高脂血症小鼠模型和HepG2细胞红色荧光标记低密度脂蛋白(Dil-LDL)摄取法考察补骨脂素对模型小鼠血脂水平和HepG2细胞Dil-LDL摄取的影响。C57BL/6J 50只小鼠适应性喂养1周,未造模小鼠作为正常组,造模后的高脂血症小鼠随机分为模型组和20、40、80 mg/kg补骨脂素组,每组10只。全自动生化仪检测丙氨酸转氨酶(alanine aminotransferase,ALT)和天冬氨酸转氨酶(aspartate aminotransferase,AST)水平,以及总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high densit lipoprotein cholesterol,HDL-C)和低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)水平。20μg/mL Dil-LDL观察补骨脂素对HepG2细胞脂质摄取的影响,分为对照组和10、20、40、80μmol/L补骨脂素组。酶标仪检测各组荧光强度;Real-time PCR和Western blot法检测补骨脂素对PCSK9和低密度脂蛋白受体(low-density lipoprotein receptor,LDLR)基因和蛋白表达的影响。结果分子对接结果显示补骨脂素与PCSK9对接结合能为-6.04 kcal/mol,提示补骨脂素可能与PCSK9结合。进一步在在体动物模型验证中发现,与高脂饮食诱导的模型组相比,20 mg/kg补骨脂素可显著降低高脂血症小鼠血清中ALT、TC和LDL-C水平(P<0.05),40 mg/kg补骨脂素显著降低高脂血症小鼠血清ALT、AST、TC、TG和LDL-C水平(P<0.05或P<0.01)。细胞实验结果显示,与对照组相比,10μmol/L、20μmol/L和40μmol/L补骨脂素可显著增强HepG2细胞Dil-LDL摄取能力(P<0.01)。与对照组相比,40μmol/L补骨脂素可显著提高LDLR mRNA和蛋白表达(P<0.05),并降低PCSK9 mRNA和蛋白表达水平(P<0.01)。结论补骨脂素具有抗高脂�
Objective To investigate the effects of psoralen on hyperlipidemia and its mechanism based on proprotein convertase subtilisin/kexin type 9(PCSK9).Methods Firstly,the molecular docking method was used to predict the binding ability of psoralen to PCSK9 protein.Subsequently,the effects of psoralen on blood lipid levels and Dil-LDL uptake in HepG2 cells were investigated by a mouse model of hyperlipidemia induced by a high-fat diet and a red fluorescently labeled probe low-density lipoprotein cholesterol(Dil-LDL)uptake in HepG2 cells,respectively.Fifty C57BL/6J mice were adaptively fed for one week,with non-modeled mice serving as the normal group.After modeling,hyperlipidemic mice were randomly divided into a model group and 20,40,and 80 mg/kg psoralen groups,with 10 mice in each group.The levels of alanine aminotransferase(ALT),and aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)were measured using an automatic biochemistry analyzer.The effects of psoralen on HepG2 cell lipid uptake were observed at 20μg/mL Dil-LDL,and the cells were divided into control group and 10,20,40,and 80μmol/L psoralen groups.And the fluorescence intensity was measured using an enzyme-linked immunosorbent assay(ELISA).Real-time PCR and Western blot were used to detect the effects of psoralen on the expression of PCSK9 and low-density lipoprotein receptor(LDLR)genes and proteins.Results The molecular docking results show that psoralen has a binding affinity of-6.04 kcal/mol with PCSK9,suggesting that psoralen may bind to PCSK9.Further in vivo animal model validation showed that compared with the model group induced by high-fat diet,20 mg/kg psoralen significantly lowered the levels of ALT,TC,and LDL-C in the serum of hyperlipidemic mice(P<0.05),and 40 mg/kg psoralen significantly lowered the levels of ALT,AST,TC,TG,and LDL-C in the serum of hyperlipidemic mice(P<0.05 or P<0.01).The results of cell experiments showed that comp
作者
刘婷
罗雅歌
毛浩萍
马尚伟
LIU Ting;LUO Yage;MAO Haoping;MA Shangwei(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300143,China)
出处
《辽宁中医药大学学报》
CAS
2024年第12期23-28,共6页
Journal of Liaoning University of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(82174180)
天津市教委科研计划项目一般项目(2022KJ182)。
