摘要
目的:探讨荷叶碱对泡沫细胞形成的影响及其可能的分子机制。方法:体外培养人单核-巨噬细胞系THP-1,利用氧化型低密度脂蛋白(Ox-LDL)诱导建立泡沫细胞模型,同时用5、10和20μmol/L荷叶碱进行干预。采用油红O染色观察细胞内脂质沉积;用试剂盒测定细胞总胆固醇含量;通过Western blot法检测微管相关蛋白1轻链3(LC3)、P62、磷酸化蛋白激酶B(p-Akt)和磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)蛋白水平,免疫荧光检测自噬流,透射电镜检测自噬小体,评估细胞内自噬水平及自噬相关通路的变化。利用自噬抑制剂3-甲基腺嘌呤(3-MA)抑制细胞自噬,观察荷叶碱是否通过调控自噬抑制泡沫细胞形成。结果:与正常对照组相比,Ox-LDL组细胞内脂质沉积和总胆固醇含量显著增加。与Ox-LDL组相比,荷叶碱组细胞内脂质沉积和总胆固醇含量显著减少,自噬流显著增强,自噬小体增多;3-MA处理后,荷叶碱抑制细胞泡沫化的作用被削弱。荷叶碱降低p-mTOR和p-Akt蛋白水平。结论:荷叶碱可能通过抑制磷脂酰肌醇3-激酶(PI3K)/Akt/mTOR通路促进自噬,减少细胞内脂质沉积,从而抑制泡沫细胞形成。
AIM:To investigate the effect of nuciferine(NUF)on the formation of foam cells and its possible molecular mechanism.METHODS:Human monocyte-macrophage cell line THP-1 was induced by oxidized low-density lipoprotein(Ox-LDL)to establish foam cell model,and simultaneously treated with NUF at 5,10 or 20μmol/L.Oil red O staining was used and total cholesterol content was measured to observe the effect of NUF on foam cell formation.Autophagy flow was detected by immunofluorescence,and autophagosomes were detected by transmission electron microscopy.The protein levels of microtubule-associated protein 1 light chain 3(LC3),P62,phosphorylated protein kinase B(p-Akt)and phosphorylated mammalian target of rapamycin(p-mTOR)were determined by Western blot.3-Methyladenine(3-MA),an autophagy inhibitor,was used to inhibit autophagy and to observe whether NUF inhibited foam cell formation by regulating autophagy.RESULTS:Compared with control group,the intracellular lipid deposition and total cholesterol content in Ox-LDL group were increased.Compared with Ox-LDL group,the intracellular lipid deposition and total cholesterol content in NUF group were decreased,while autophagy flow and number of autophagosomes were increased.The inhibitory effect of NUF on cell foaming was weakened after 3-MA treatment.Moreover,NUF decreased the protein levels of p-mTOR and p-Akt.CONCLUSION:Nuciferine may promote autophagy by inhibiting phosphatidylinositol 3-kinase(PI3 K)/Akt/mTOR signaling pathway,thus reducing intracellular lipid deposition and formation of foam cells.
作者
丁畅
银萍
赵奇
苏立
DING Chang;YIN Ping;ZHAO Qi;SU Li(Department of Cardiology,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400014,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第7期1230-1236,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81974036)。