天然产物研究与开发 ›› 2021, Vol. 33 ›› Issue (7): 1178-1185.doi: 10.16333/j.1001-6880.2021.7.013

所属专题: No.6

• 开发研究 • 上一篇    下一篇

绞股蓝皂苷调控长链非编码RNA TUG1/miR-26a干扰线粒体凋亡对ApoE-/-AS小鼠肝脏脂质沉积的影响及机制研究

宋囡1,2,曹慧敏1,2,陈丝1,2,王莹1,2,王杰1,2,王群1,2,贾连群1,2*,杨关林1,2*   

  1. 1辽宁中医药大学 中医药创新工程技术中心 脏象理论及应用教育部重点实验室;2辽宁中医药大学,沈阳 110847
  • 出版日期:2021-07-28 发布日期:2021-07-29
  • 基金资助:
    中医药传承与创新“百千万”人才工程(岐黄工程);辽宁省自然科学基金指导计划项目(2019-ZD-0966);辽宁省“兴辽英才计划”项目(XLYC1902100)

 Gypenoside regulates long non-coding RNA TUG1/miR-26a by interfering with mitochondrial apoptosis on hepatic lipid deposition of ApoE-/-AS mice

SONG Nan1,2,CAO Hui-min1,2,CHEN Si1,2,WANG Ying1,2, WANG Jie1,2,WANG Qun1,2,JIA Lian-qun1,2*,YANG Guan-lin1,2*   

  1. 1Chinese Medicine Innovation Engineering Technology Center,Liaoning Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,University of Traditional Chinese Medicine;2Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China

  • Online:2021-07-28 Published:2021-07-29

摘要:

为探讨绞股蓝皂苷通过影响长链非编码RNA TUG1/miR-26a干扰线粒体凋亡改善ApoE-/-AS小鼠肝脏脂质沉积防治AS机制,本实验将10只C57BL/6J小鼠作为正常对照组,20只健康ApoE-/-小鼠随机分为模型组、绞股蓝皂苷组(高脂饲料喂养12周),灌胃给药4周。HE染色观察小鼠肝脏脂质沉积情况,全自动生化分析仪检测血脂水平,实时荧光定量Q-PCR检测长链非编码TUG1、miRNA-26a表达,实时荧光定量Q-PCR及Wes全自动蛋白质印迹定量分析系统检测Bcl2、Bax、Cytc、cleaved caspase-3、cleaved caspase-9、cleaved PARP基因及 蛋白表达。结果显示模型组ApoE-/-小鼠血脂水平发生紊乱,肝细胞体积变大,脂肪空泡明显,小鼠肝脏Lnc-TUG1表达显著升高,miRNA-26a显著下降(P<0.01);Bax、Cyt-c、cleaved caspase-3、cleaved PARP mRNA及蛋白表达显著升高,Bcl2 mRNA及蛋白显著下降(P<0.01或P<0.05);cleaved caspase-9 蛋白表达显著升高(P<0.01或P<0.05),cleaved caspase-9 mRNA仅有上升趋势;绞股蓝皂苷干预后血脂紊乱得以改善,肝细胞脂肪变性程度减轻,脂肪空泡明显减少,小鼠肝脏Lnc TUG1表达有所下降,miRNA-26a 表达有所上调(P<0.05),小鼠肝脏Bax、Cyt-c、cleaved caspase-3、cleaved caspase-9 mRNA及蛋白表达显著下调,Bcl2 mRNA及蛋白显著上调(P<0.01或P<0.05),cleaved PARP蛋白表达显著下调(P<0.05),cleaved PARP mRNA仅有下调趋势;研究结果提示绞股蓝皂苷可能通过影响长链非编码RNA TUG1/miR-26a干扰线粒体凋亡改善ApoE-/-AS小鼠肝脏脂质沉积,进而防治动脉粥样硬化。

关键词: 绞股蓝皂苷, 动脉粥样硬化, 长链非编码RNA TUG1/miR-26a, 肝脏脂质沉积, 线粒体凋亡

Abstract:

To explore the mechanism of gypenoside preventing and treating AS by affecting long non-coding RNA TUG1/miR-26a to interfere with mitochondrial apoptosis,thereby improving liver lipid deposition in ApoE-/-AS mice.In this experiment,10 C57BL/6J mice were used as the normal control group,and 20 healthy ApoE-/-mice fed with high-fat diet for 12 weeks were randomly divided into model group and gypenoside group,given intragastrically for 4 weeks.Lipid deposition in mouse liver was observed by HE staining,blood lipid level was detected by automatic biochemical analyzer,and mRNA expression of long non-coding TUG1,miRNA-26a,Bcl2,Bax,Cytc,cleaved caspase-3,cleaved caspase-9 and cleaved PARP were detected by real-time q-PCR,and protein expression of Bcl2,Bax,Cytc,cleaved caspase-9 and cleaved PARP were detected by Wes automatic Western blotting quantitative analysis system.The results showed that the blood lipid level of ApoE-/- mice in the model group was disordered,liver cell volume increased,and fat vacuoles were obvious.The expression of Lnc-TUG1 in mouse liver was significantly increased,and miRNA-26a was significantly decreased (P<0.01);Bax,Cytc,cleaved caspase-3,cleaved PARP mRNA and protein expression significantly increased,Bcl2 mRNA and protein expression significantly decreased P<0.01 or P<0.05);cleaved caspase-9 protein expression significantly increased(P<0.05),cleaved caspase-9 mRNA only has an upward trend.After the intervention of gypenoside,dyslipidemia was improved,the degree of liver cell steatosis was reduced,fatty vacuoles were significantly reduced.Lnc-TUG1 expression was decreased,miRNA-26a expression was increased (P<0.05),the mRNA and protein expressions of Bax,Cytc,cleaved caspase-3 and cleaved caspase-9 were significantly down-regulated,Bcl2 mRNA and protein was significantly up-regulated,the expressions of cleaved PARP protein were significantly down-regulated(P<0.01 or P<0.05),and cleaved PARP mRNA showed only a downregulation trend.The results suggested that the effect of gypenoside in preventing and treating atherosclerosis may be to interfere with mitochondrial apoptosis by affecting long non-coding RNA TUG1/miR-26a,thereby improving liver lipid deposition in ApoE-/-AS mice.

Key words: gypenoside, atherosclerosis, long chain non-coding RNA TUG1/ miR-26a, hepatic lipid deposition, mitochondrial apoptosis

中图分类号:  R285.5