中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (7): 1080-1084.doi: 10.12307/2022.148

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

大蒜素抑制大鼠血管内皮细胞增殖并诱导其凋亡的机制

张玉杰1，2，杨建东2 ，蔡  俊2 ，朱守雷2 ，田  原2   

  1. 1大连医科大学研究生院，辽宁省大连市  116044；2苏北人民医院脊柱科，江苏省扬州市  225001
  • 收稿日期:2020-09-26 修回日期:2020-09-30 接受日期:2020-11-13 出版日期:2022-03-08 发布日期:2021-10-29
  • 通讯作者: 杨建东，博士，副教授，硕士生导师，苏北人民医院脊柱科，江苏省扬州市 225001
  • 作者简介:张玉杰，男，1995年生，安徽省太和县人，汉族，大连医科大学外科学(骨科方向) 在读硕士，主要从事骨科各学科临床治疗及基础研究。
  • 基金资助:
    江苏省医学创新团队(CXTD2017004)，项目参与人：杨建东；扬州市“十三五”科教强卫专项经费(LJRC20182)，项目负责人：杨建东

Mechanism by which allicin inhibits proliferation and promotes apoptosis of rat vascular endothelial cells

Zhang Yujie1, 2, Yang Jiandong2, Cai Jun2, Zhu Shoulei2, Tian Yuan2   

  1. 1Graduate School of Dalian Medical University, Dalian 116044, Liaoning Province, China; 2Department of Spine, North Jiangsu People’s Hospital, Yangzhou 225001, Jiangsu Province, China
  • Received:2020-09-26 Revised:2020-09-30 Accepted:2020-11-13 Online:2022-03-08 Published:2021-10-29
  • Contact: Yang Jiandong, MD, Associate professor, Master’s supervisor, Department of Spine, North Jiangsu People’s Hospital, Yangzhou 225001, Jiangsu Province, China
  • About author:Zhang Yujie, Master candidate, Graduate School of Dalian Medical University, Dalian 116044, Liaoning Province, China; Department of Spine, North Jiangsu People’s Hospital, Yangzhou 225001, Jiangsu Province, China
  • Supported by:
    the Jiangsu Provincial Medical Innovation Team, No. CXTD2017004 (to YJD); Yangzhou Health Bureau’s Leading Talent of Science, Education Strengthening Health of Yangzhou during the 13th Five-Year Plan Period, No. LJRC20182 (to YJD)

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摘要：

文题释义：
大蒜素：是新鲜大蒜组织破碎时产生的一种具有典型气味的天然含硫化合物。作为一种硫代亚磺酸盐，大蒜素能与谷胱甘肽和蛋白质中的硫醇基团发生氧化还原反应，从而发挥多种不同的生物学特性。在微生物、植物和哺乳动物细胞中，大蒜素能以剂量依赖性方式抑制细胞增殖并诱导细胞死亡。在亚致死浓度下，大蒜素具有多种健康促进特性。
血管内皮细胞：是覆盖动脉、毛细血管和静脉内壁最内部的结构，是血液和组织之间的屏障，在调节免疫反应、炎症和血管生成中发挥重要作用。正常生理条件下，血管内皮细胞保持静止状态，当出现组织损伤或缺氧时，血管内皮细胞高度协调迅速形成新血管。
背景：大蒜素具有抗纤维化和抗血管生成作用，既往研究已经证实了大蒜素对成纤维细胞过度增殖的抑制作用，但大蒜素对于血管内皮细胞增殖和凋亡的影响及其确切机制尚未明确。
目的：观察大蒜素对血管内皮细胞形态、增殖和凋亡的影响及其可能机制，为大蒜素下一步临床应用提供理论基础。
方法：血管内皮细胞常规培养至对数期，按照大蒜素质量浓度分为对照组(0 mg/L)、低质量浓度组(25 mg/L)、中质量浓度组(50 mg/L)和高质量浓度组(100 mg/L)，干预24 h后使用CCK-8法检测细胞活力，倒置相差显微镜观察细胞形态变化，AO/EB双重染色法和AnnexinV-FITC双重染色法检测细胞凋亡率，PI/RNase法检测细胞周期分布，Western blot和RT-PCR检测PCNA、Bax、BcL-2蛋白和基因的表达水平。
结果与结论：用0，25，50，100 mg/L大蒜素处理24 h后，血管内皮细胞的活力以时间剂量依赖性方式显著降低，24 h IC50值为
103.27 mg/L。随着大蒜素质量浓度的增加，血管内皮细胞的凋亡率上升(P < 0.01)； G1期细胞数减少(P < 0.01)，G2期细胞数增加(P < 0.01)；与对照组相比，大蒜素组PCNA和Bcl-2的表达降低(P < 0.01)，而Bax的表达显著升高(P < 0.01)。结果表明，大蒜素具有抑制血管内皮细胞增殖并促其凋亡的作用，作用机制可能与调控PCNA、Bcl-2及Bax表达有关。

https://orcid.org/0000-0002-1053-2057(杨建东) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


关键词: 血管内皮细胞, 大蒜素, 增殖, 凋亡, 因子, 机制, 硬膜外瘢痕

Abstract: BACKGROUND: Allicin has anti-fibrosis and anti-angiogenesis effects. Previous studies have confirmed the inhibitory effect of allicin on the proliferation of fibroblasts, but the effect and exact mechanism of allicin on the proliferation and apoptosis of vascular endothelial cells are still unclear.
OBJECTIVE: To observe the effect of allicin on the morphology, proliferation and apoptosis of vascular endothelial cells and its possible mechanism, and provide a theoretical basis for the next clinical application of allicin.  
METHODS: Vascular endothelial cells were routinely cultured to logarithmic phase in vitro, and the cells were divided into control group (0 mg/L), low concentration group (25 mg/L), medium concentration group (50 mg/L), and high concentration group (100 mg/L) according to the concentration of allicin in the medium. After 24 hours of intervention, CCK-8 assay was used to detect cell viability. Inverted phase contrast microscope was used to observe cell morphological changes. AO/EB double staining method and AnnexinV-FITC double staining method were used to detect apoptosis rate. PI/RNase method was used to detect cell cycle. Reverse-transcription PCR and western blot assay were used to measure the expression levels of PCNA, Bax and BcL-2 proteins and genes.
RESULTS AND CONCLUSION: After treated with allicin at concentrations of 0, 25, 50 and 100 mg/L for 24 hours, the viability of vascular endothelial cells significantly decreased in a time-dose-dependent manner, with an IC50 value of 103.27 mg/L at 24 hours. As the concentrations of allicin increased, the apoptosis rate of vascular endothelial cells rose up (P < 0.01); the cell numbers at G1 phase decreased (P < 0.01); and at G2 phase increased (P < 0.01). Compared with control group, the expression of PCNA and Bcl-2 decreased (P < 0.01), while the expression of Bax increased significantly in the allicin group (P < 0.01). The results suggested that allicin inhibited the proliferation of vascular endothelial cells and induced their apoptosis. The action mechanism may be achieved by regulating the expression of PCNA, Bcl-2 and Bax.

Key words: vascular endothelial cells, allicin, proliferation, apoptosis, factor, mechanism, epidural scar

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