College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
BMC Complement Altern Med. 2014 Aug 30;14:321. doi: 10.1186/1472-6882-14-321.
Allicin, a primary ingredient of garlic, has been proposed to possess cardioprotective properties, which are commonly mediated by improved endothelial function.
To investigate the effect and mechanism of allicin on the apoptosis of human umbilical vein endothelial cells (HUVECs), we used Propidium iodide (PI) staining and Annexin V/ PI staining assays to establish a model of oxidative stress apoptosis induced by H2O2. MTT, RT-PCR and western-blot assays were used to detect the effects and mechanism of allicin on the model.
PI staining, Annexin V/ PI staining assays and morphological assessment suggest that the cell death induced by 0.5 mM H2O2 is primarily apoptotic. Conversely, allicin reverses the effect of H2O2 on cell death, suggesting a role in protecting HUVECs from apoptosis. We demonstrated that H2O2 activates PARP cleavage, reduces pro-Caspase-3 levels and activates Bax expression; however, allicin inhibits each of these apoptotic signaling indicators. Allicin also reduces the levels of malondialdehyde and increases the levels of superoxide dismutase, nitric oxide release and endothelial nitric oxide synthase mRNA, but has no significant effect on inducible nitric oxide synthase mRNA levels.
These results demonstrate that allicin has powerful effects in protecting HUVECs from apoptosis and suggest that protection occurs via a mechanism involving the protection from H2O2-mediated oxidative stress.
大蒜中的主要成分蒜素被认为具有心脏保护作用,这通常是通过改善内皮功能来实现的。
为了研究蒜素对人脐静脉内皮细胞(HUVEC)凋亡的影响及其机制,我们使用碘化丙啶(PI)染色和 Annexin V/PI 染色试验建立了由 H2O2 诱导的氧化应激凋亡模型。MTT、RT-PCR 和 Western blot 试验用于检测蒜素对模型的作用和机制。
PI 染色、Annexin V/PI 染色试验和形态学评估表明,0.5mM H2O2 诱导的细胞死亡主要是凋亡。相反,蒜素逆转了 H2O2 对细胞死亡的影响,表明其在保护 HUVEC 免于凋亡中的作用。我们证明 H2O2 激活 PARP 裂解,降低前胱冬酶-3 水平并激活 Bax 表达;然而,蒜素抑制了这些凋亡信号指标的每一个。蒜素还降低丙二醛水平,增加超氧化物歧化酶、一氧化氮释放和内皮型一氧化氮合酶 mRNA 的水平,但对诱导型一氧化氮合酶 mRNA 水平没有显著影响。
这些结果表明,蒜素在保护 HUVEC 免于凋亡方面具有强大的作用,并表明保护作用是通过一种涉及保护 H2O2 介导的氧化应激的机制来实现的。