大蒜素抑制大鼠血管内皮细胞增殖并诱导其凋亡的机制  

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

作　　者：张玉杰 杨建东 蔡俊 朱守雷 田原 Zhang Yujie;Yang Jiandong;Cai Jun;Zhu Shoulei;Tian Yuan(Graduate School of Dalian Medical University,Dalian 116044,Liaoning Province,China;Department of Spine,North Jiangsu People’s Hospital,Yangzhou 225001,Jiangsu Province,China)

机构地区：[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

出　　处：《中国组织工程研究》2022年第7期1080-1084,共5页Chinese Journal of Tissue Engineering Research

基　　金：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。

摘　　要：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 IC_(50)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);G_(1)期细胞数减少(P<0.01),G_(2)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。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 IC_(50) 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 G_(1) phase decreased(P<0.01);and at G_(2) 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 regulatin

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 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]