文章摘要
王翅遥,高严格,张晓东,康晓军,陈蕊蕊.葛根素延缓动脉粥样硬化进展的机制研究[J].,2020,(16):3022-3027
葛根素延缓动脉粥样硬化进展的机制研究
Study on the Mechanism of Puerarin in Preventing the Progression of Atherosclerosis
投稿时间:2020-03-03  修订日期:2020-03-28
DOI:10.13241/j.cnki.pmb.2020.16.005
中文关键词: 葛根素  动脉粥样硬化  Sirt3  凋亡  巨噬细胞
英文关键词: Puerarin  Atherosclerosis  Sirt3  Apoptosis  Macrophages
基金项目:国家自然科学基金青年项目(81800346);陕西省自然科学基础研究计划-一般项目(2019JQ-422,808081369018)
作者单位E-mail
王翅遥 空军军医大学唐都医院心内科 陕西 西安 710032 408943506@qq.com 
高严格 长庆油田职工医院心内科 陕西 西安710200  
张晓东 西安国际医学中心 陕西 西安 710100  
康晓军 西安医学院第二附属医院心内科 陕西 西安 710038  
陈蕊蕊 空军军医大学唐都医院心内科 陕西 西安 710032  
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中文摘要:
      摘要 目的:明确葛根素(Pur)对小鼠动脉粥样硬化斑块稳定性的治疗作用及其潜在机制。方法:用高脂饮食喂养成年小鼠诱导动脉粥样硬化模型,将小鼠分为Con组,ApoE-/-组,ApoE-/-+ Pur组。采用HE染色分别检测各组小鼠中动脉粥样硬化斑块面积,免疫荧光染色检测斑块中MMP2的阳性区域面积。用50 μg/mL ox-LDL干预巨噬细胞诱导动脉粥样硬化细胞模型,并用Ad-sh-Sirt3干扰Sirt3表达,将细胞分为Con组、ox-LDL组、ox-LDL+Pur组、ox-LDL+Pur+Ad-sh-Sirt3组。Western-blot检测Sirt3表达含量,TUNEL法检测巨噬细胞凋亡。结果:动物水平,与Con组相比,ApoE-/-组小鼠出现了显著的动脉粥样硬化斑块,ApoE-/-组小鼠MMP2阳性区域面积显著高于Con组(P<0.05);Pur处理后,ApoE-/-+ Pur组动脉粥样硬化斑块面积明显低于ApoE-/-组(P<0.05),MMP2的阳性区域面积显著低于ApoE-/-组(P<0.05)。细胞水平,Western结果显示,与对照组相比,ox-LDL组Sirt3表达量显著降低(P<0.05),ox-LDL+Pur组Sirt3表达水平显著高于ox-LDL组(P<0.05)。相比于Con组,ox-LDL组巨噬细胞凋亡水平显著升高(P<0.05);给予Pur处理后,相比于单纯ox-LDL组,ox-LDL+Pur组巨噬细胞的凋亡水平显著降低(P<0.05)。Ad-sh-Sirt3处理消除了葛根素对于巨噬细胞凋亡的抑制作用(P<0.05)。结论:外源性Pur可能通过激活Sirt3表达,进一步降低巨噬细胞凋亡水平,减少巨噬细胞的浸润,增加动脉粥样硬化斑块稳定性。
英文摘要:
      ABSTRACT Objective: To clarify the therapeutic effect of Puerarin (Pur) on stability of atherosclerotic plaques and its underlying mechanism. Methods: High-fat-diet was used to induce atherosclerosis model, the mice were divided into Con group, ApoE-/- group and ApoE-/-+Pur group. The plaque area was measured by HE staining, the positive area of MMP2 in plaques were detected by immunofluorescence staining. In vitro, 50 μg/mL ox-LDL was used to simulate atherosclerosis and Ad-sh-Sirt3 was used to inhibit Sirt3 expression. Cells were divided into Con group, ox-LDL group, ox-LDL + Pur group, ox-LDL + Pur + Ad-sh-Sirt3 group. Western-blot was used to detect the expression of Sirt3. The level of apoptosis in macrophages was detected by TUNEL assay. Results: Compared with the Con group, the mice in the ApoE-/- group showed significant atherosclerotic plaques, the positive area of MMP2 in ApoE-/- mice were significantly higher than Con group(P<0.05). The plaque area and the positive area of MMP2 in ApoE-/- mice were significantly higher than ApoE-/-+Pur group (P<0.05). In cell level, western blot showed that compared with Con group, the expression of Sirt3 in ox-LDL group was significantly reduced (P<0.05). Treatment with Pur significantly increased the expression of Sirt3 compared with ox-LDL group (P<0.05). Furthermore, compared with the Con group, the level of macrophage apoptosis with ox-LDL administration was significantly increased (P<0.05), and Pur administration significantly reduced the apoptosis levels of cells compared with ox-LDL group (P<0.05). Conclusion: Pur effectively reduced the level of macrophage apoptosis and the infiltration of macrophages, increased the stability of atherosclerotic plaques by activating Sirt3 expression.
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