Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan, 250012, PR China.
Pharmacol Rep. 2011;63(6):1469-80. doi: 10.1016/s1734-1140(11)70711-3.
A novel stilbene derivative, (E)-2-(2-chlorostyryl)-3,5,6-trimethylpyrazine (CSTMP), was designed and synthesized based on the pharmacophores of tetramethylpyrazine (TMP) and resveratrol (RES). In the present study, we investigated the protective effects of CSTMP on vascular endothelial cells under oxidative stress and elucidated its molecular mechanisms. The radical scavenging activity of CSTMP was assessed by the DPPH test. Human Umbilical Vein Endothelial Cells (HUVECs) were exposed to 150 μM hydrogen peroxide (H(2)O(2)) for 12 h, resulting in a decrease of cell viability assessed by the MTT assay and an increase of apoptotic cells assessed by the nuclear staining assay and flow cytometry. The activities of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and nitric oxide synthase (NOS) and the contents of malondialdehyde (MDA), reduced glutathione (GSH) and nitric oxide (NO) in cells were determined by commercial kits. The expression levels of pro-apoptotic factor caspase-3 and anti-apoptotic signal ERK1/2 were detected by western blot. The results showed that CSTMP had a moderate anti-oxidative effect against the DPPH test, which was less than RES. Co-incubation with CSTMP increased the cell viability, markedly reduced the LDH leakage from the cells and decreased the lipid peroxidation. These effects of CSTMP were accompanied by increasing activity of the endogenous antioxidant enzyme SOD, the level of GSH, the production of NO and cNOS activity. Moreover, CSTMP showed stronger effects on the inhibition of apoptosis, caspase-3 expression, and the activation of phosphorylated ERK1/2 compared to RES. Furthermore, CSTMP could inhibit the expression of phospho-JNK and phospho-p38 induced by H(2)O(2). These results suggest that CSTMP prevents H(2)O(2)-induced cell injury through anti-oxidation and anti-apoptosis via the MAPK and caspase-3 pathways.
一种新型的二苯乙烯衍生物，（E）-2-（2-氯苯乙烯基）-3,5,6-三甲基吡嗪（CSTMP），是基于四甲基吡嗪（TMP）和白藜芦醇（RES）的药效团设计和合成的。在本研究中，我们研究了 CSTMP 在氧化应激下对血管内皮细胞的保护作用，并阐明了其分子机制。CSTMP 的自由基清除活性通过 DPPH 试验进行评估。用 150μM 过氧化氢（H2O2）处理人脐静脉内皮细胞（HUVEC）12 小时，通过 MTT 测定法评估细胞活力下降，通过核染色法和流式细胞术评估凋亡细胞增加。通过商业试剂盒测定乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）和一氧化氮合酶（NOS）的活性以及丙二醛（MDA）、还原型谷胱甘肽（GSH）和一氧化氮（NO）的含量。通过 Western blot 检测促凋亡因子 caspase-3 和抗凋亡信号 ERK1/2 的表达水平。结果表明，CSTMP 对 DPPH 试验具有中等的抗氧化作用，弱于 RES。与 CSTMP 共孵育可提高细胞活力，显著减少细胞内 LDH 的渗漏，并降低脂质过氧化。CSTMP 的这些作用伴随着内源性抗氧化酶 SOD 活性的增加、GSH 水平的升高、NO 的产生和 cNOS 活性的增加。此外，与 RES 相比，CSTMP 对抑制细胞凋亡、caspase-3 表达和磷酸化 ERK1/2 的激活具有更强的作用。此外，CSTMP 可以抑制 H2O2 诱导的 JNK 和 p38 的磷酸化表达。这些结果表明，CSTMP 通过 MAPK 和 caspase-3 途径预防 H2O2 诱导的细胞损伤，从而发挥抗氧化和抗凋亡作用。
