Baotou Medical College, Baotou 014000, China.
Inner Mongolia Autonomous Region Hospital of Traditional Chinese Medicine, Hohhot 010020, China.
Biomed Res Int. 2019 Jan 20;2019:2692970. doi: 10.1155/2019/2692970. eCollection 2019.
In previous studies, (Michx.) Hulten was reported to contain xanthones, iridoids, terpenoids, and sterols and is mainly used to cure hepatitis, jaundice, fever, headache, and angina pectoris. In this study, we used bioassay guided fractionation to identify compounds from and investigated their activity against hydrogen peroxide (HO)-induced apoptosis of H9c2 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic (GCLC) expression were assessed using quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was evaluated using western blot. The results showed that all four compounds had protective effects on H9c2 cells. The transcription levels of HO-1 and GCLC significantly increased in H9c2 cells pretreated with norswertianolin (), swetrianolin (), demethylbellidifolin (), and bellidifolin (). However, compared to the model group, the transcription levels of Nrf2 were not enhanced by pretreatment with compounds , , and . The protein expression levels of HO-1 and GCLC in H9c2 cells were greater than that in the HO-treated group, and the expression of Nrf2 was not significantly changed except by swetrianolin treatment; inhibitors can reverse the protective effect by ZnPP (15 M), BSO (10 M), and brusatol (10 M). The results indicated that the four compounds isolated from inhibited the oxidative injury induced by HO by activating the Nrf2/ARE pathway in H9c2 cells and provide evidence that may be a potential therapeutic agent for the treatment of cardiovascular diseases.
在之前的研究中，报道（Michx.）Hulten 含有黄烷酮、环烯醚萜、萜类和甾体，并主要用于治疗肝炎、黄疸、发热、头痛和心绞痛。在这项研究中，我们使用生物测定指导的分级分离来鉴定来自的化合物，并使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法研究它们对过氧化氢（HO）诱导的 H9c2 细胞凋亡的活性。核因子红细胞 2 相关因子 2（Nrf2）、血红素加氧酶-1（HO-1）和谷氨酸半胱氨酸连接酶催化亚基（GCLC）的表达水平使用定量实时聚合酶链反应（qRT-PCR）进行评估。使用蛋白质印迹评估蛋白质表达。结果表明，所有四种化合物对 H9c2 细胞均具有保护作用。HO-1 和 GCLC 的转录水平在用 norswertianolin（）、swetrianolin（）、demethylbellidifolin（）和 bellidifolin（）预处理的 H9c2 细胞中显著增加。然而，与模型组相比，用化合物预处理后，Nrf2 的转录水平没有增强， ， 。HO-1 和 GCLC 在 H9c2 细胞中的蛋白表达水平均大于 HO 处理组，除 swetrianolin 处理外，Nrf2 的表达没有明显变化；抑制剂可以通过 ZnPP（15 μM）、BSO（10 μM）和 brusatol（10 μM）逆转保护作用。结果表明，从 中分离出的四种化合物通过激活 H9c2 细胞中的 Nrf2/ARE 途径抑制 HO 诱导的氧化损伤，并为 可能是治疗心血管疾病的潜在治疗剂提供证据。
