ACS6, a Hydrogen sulfide-donating derivative of sildenafil, inhibits homocysteine-induced apoptosis by preservation of mitochondrial function.
ACS6, a Hydrogen sulfide-donating derivative of sildenafil, inhibits homocysteine-induced apoptosis by preservation of mitochondrial function.
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DOI：
10.1186/2045-9912-1-20
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发表时间：
2011-08-16
影响因子：
2.9
通讯作者：
Wang CY
中科院分区：
文献类型：
--
作者：
Tang XQ;Chen RQ;Ren YK;Soldato PD;Sparatore A;Zhuang YY;Fang HR;Wang CY
释放硫化氢的西地那非 (ACS6) 已被证明可以通过释放硫化氢 (H2S) 来抑制超氧化物的形成。我们发现 H2S 可以拮抗同型半胱氨酸诱导的氧化应激和神经毒性。本研究的目的是探讨 ACS6 对同型半胱氨酸引发的细胞毒性和细胞凋亡的保护作用及其在 PC12 细胞中的分子机制。通过 Cell Counting Kit-8 测定法测定细胞活力。使用染色质染料 Hoechst 33258 观察细胞凋亡，并在碘化丙啶染色后通过流式细胞术进行分析。使用荧光染料 Rh123 监测线粒体膜电位。通过细胞渗透性 2',7'-二氯荧光素二乙酸酯氧化转化为荧光 2',7'-二氯荧光素来测定细胞内活性氧。通过Western blot分析cleaved caspase-3和bcl-2的表达以及胞质细胞色素c的积累。我们发现 ACS6 可以保护 PC12 细胞免受同型半胱氨酸诱导的细胞毒性和细胞凋亡，并阻断同型半胱氨酸触发的细胞色素 c 释放和 caspase-3 激活。 ACS6 治疗不仅可以预防同型半胱氨酸引起的线粒体膜电位 (Δψ) 损失和活性氧 (ROS) 过量产生，还可以逆转 Bcl-2 的下调表达。这些结果表明，ACS6 通过抑制 Δψ 的损失和 ROS 的积累以及调节 Bcl-2 的表达，通过保留线粒体功能来保护 PC12 细胞免受同型半胱氨酸诱导的细胞毒性和凋亡。我们的研究为 ACS6 的神经保护作用和进一步评估 ACS6 作为与同型半胱氨酸相关的阿尔茨海默病的新型神经保护剂提供了证据。
The hydrogen sulfide-releasing sildenafil, ACS6, has been demonstrated to inhibit superoxide formation through donating hydrogen sulfide (H2S). We have found that H2S antagonizes homocysteine-induced oxidative stress and neurotoxicity. The aim of the present study is to explore the protection of ACS6 against homocysteine-triggered cytotoxicity and apoptosis and the molecular mechanisms underlying in PC12 cells. Cell viability was determined by Cell Counting Kit-8 assay. Cell apoptosis was observed using the chromatin dye Hoechst 33258 and analyzed by Flow Cytometry after propidium iodide staining. Mitochondrial membrane potential was monitored using the fluorescent dye Rh123. Intracellular reactive oxygen species were determined by oxidative conversion of cell permeable 2',7'-dichlorfluorescein-diacetate to fluorescent 2',7'-dichlorfluorescein. The expression of cleaved caspase-3 and bcl-2 and the accumulation of cytosolic cytochrome c were analyzed by Western blot. We show that ACS6 protects PC12 cells against cytotoxicity and apoptosis induced by homocysteine and blocks homocysteine-triggered cytochrome c release and caspase-3 activation. ACS6 treatment results in not only prevention of homocysteine-caused mitochondrial membrane potential (Δψ) loss and reactive oxygen species (ROS) overproduction but also reversal of Bcl-2 down-expression. These results indicate that ACS6 protects PC12 cells against homocysteine-induced cytotoxicity and apoptosis by preservation of mitochondrial function though inhibiting both loss of Δψ and accumulation of ROS as well as modulating the expression of Bcl-2. Our study provides evidence both for a neuroprotective effect of ACS6 and for further evaluation of ACS6 as novel neuroprotectants for Alzheimer's disease associated with homocysteine.