The active components of Dracocephalum moldavica L. (TFDM) can inhibit myocardial ischemia by inhibiting oxidative stress. However, the effects of TFDM on astrocytes have not been investigated in vitro. The current study aimed to explore whether TFDM protects astrocytes against H2O2-induced apoptosis through a mitochondria-dependent pathway. The human glioma cell line U87 was used to investigate the ability of TFDM to protect astrocytes against H2O2-induced apoptosis. The cell counting kit-8 assay and flow cytometry were used to detect cell viability, apoptosis, MMP, Ca2+ influx and reactive oxygen species (ROS). Lactate dehydrogenase (LDH) and malonic dialdehyde (MDA) levels were measured by ELISA. In addition, protein and mRNA expression changes were detected by Western blotting and qRT-PCR. TFDM (0.78~200 μg/ml) had limited cytotoxic effects on the viability of U87 cells. Compared with the model group (treated with H2O2 only), cells treated with medium- and high-dose TFDM exhibited reduced MDA concentrations (P < 0.05) and ROS production (P < 0.05) and decreased MMP (P < 0.05) and reduced apoptosis (P < 0.05). The percentage of annexin V-FITC-stained cells was markedly suppressed by TFDM, confirming its anti-apoptotic properties. WB results showed that protein expression of Bcl-2-associated X protein (BAX), Caspase-3, Caspase-9, Caspase-12, and B-cell leukemia/lymphoma 2 (Bcl2) was reduced in the TFDM group compared with that in the model group (P < 0.05) and that expression of these proteins was normalized by TFDM treatment in a dose-dependent manner. According to RT-qPCR results, TFDM pretreatment resulted in reduced mRNA expression of BAX, Caspase-9, Caspase-12, p38MAPK, and CaMKII and increased mRNA expression of mTOR compared with the model group. The current study revealed the protective effects of TFDM on U87 cells under oxidative stress conditions through the inhibition of a mitochondria-dependent pathway that is associated with the CaMKII/P38MAPK/ERK1/2 and PI3K/AKT/mTOR pathways.

中文翻译：

---

头孢霉菌总黄酮通过线粒体依赖性途径保护星形胶质细胞免受H2O2诱导的凋亡。

霉菌Dracocephalum Moldavica L.（TFDM）的活性成分可通过抑制氧化应激来抑制心肌缺血。但是，尚未在体外研究TFDM对星形胶质细胞的作用。当前的研究旨在探讨TFDM是否通过线粒体依赖性途径保护星形胶质细胞免受H2O2诱导的细胞凋亡。人类神经胶质瘤细胞系U87用于研究TFDM保护星形胶质细胞免受H2O2诱导的细胞凋亡的能力。使用细胞计数试剂盒8检测和流式细胞仪检测细胞活力，凋亡，MMP，Ca2 +流入和活性氧（ROS）。通过ELISA测量乳酸脱氢酶（LDH）和丙二醛（MDA）水平。另外，通过蛋白质印迹和qRT-PCR检测蛋白质和mRNA表达变化。TFDM（0。78〜200μg/ ml）对U87细胞活力的细胞毒性作用有限。与模型组（仅用H2O2处理）相比，用中，大剂量TFDM处理的细胞的MDA浓度降低（P <0.05）和ROS产生（P <0.05），MMP降低（P <0.05），凋亡减少（P ＜0.05）。膜联蛋白V-FITC染色的细胞百分比被TFDM显着抑制，证实了其抗凋亡特性。WB结果表明，与DMDM组相比，Bcl-2相关X蛋白（BAX），Caspase-3，Caspase-9，Caspase-12和B细胞白血病/淋巴瘤2（Bcl2）的蛋白表达降低。在模型组中（P ＜0.05），这些蛋白的表达通过TFDM处理以剂量依赖性方式被标准化。根据RT-qPCR结果，与模型组相比，TFDM预处理导致BAX，Caspase-9，Caspase-12，p38MAPK和CaMKII的mRNA表达降低，而mTOR的mRNA表达升高。当前的研究揭示了在氧化应激条件下，TFDM通过抑制与CaMKII / P38MAPK / ERK1 / 2和PI3K / AKT / mTOR途径相关的线粒体依赖性途径对U87细胞的保护作用。