Department of Cardiology, Affiliated Liyuan Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China.
Department of Human Anatomy and Embryology, Medical College of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.
Int J Mol Med. 2017 Dec;40(6):1889-1894. doi: 10.3892/ijmm.2017.3186. Epub 2017 Oct 12.
Puerarin extracted from Radix Puerariae is well known for its pharmacological effects, including antioxidant, anti‑inflammatory, neuroprotective and cardioprotective properties. In this study, we aimed to investigate the effects of puerarin on the daunorubicin (DNR)-induced apoptosis of H9c2 cells and to elucidate the potential mechanisms involved. MTT assay and flow cytometry were performed to evaluate cell cytotoxicity and apoptosis, respectively. Western blot analysis was used to assess changes in the expression levels of proteins, including caspase-3, Akt and phosphorylated Akt (p-Akt). Ratiometric imaging of intracellular calcium (Ca2+) using cells loaded with Fura-2 was also carried out. Our results revealed that puerarin pre-treatment protected the H9c2 cells against DNR-induced cytotoxicity by inhibiting cell apoptosis, which was also confirmed by the decrease in the expression of cleaved caspase-3. Additionally, p-Akt activation was associated with the suppressive effects of puerarin. Following pre-treatment with puerarin, the extracellular Ca2+ influx was restrained and this resulted in a reduction in the intracellular Ca2+ levels; these effects were abrogated by LY294002 [an inhibitor of phosphatidylinositol 3-kinase (PI3K)]. The inhibition of Ca2+ influx suggested that the PI3K/Akt signaling pathway participated in the suppressive effects of puerarin against H9c2 cell apoptosis. Taken togher, our findings demonstrate that puerarin attenuates the DNR-induced apoptosis of H9c2 cells by activating the PI3K/Akt signaling pathway via the inhibition of Ca2+ influx, suggesting that puerarin may be a potential cardioprotective agent for use in the clinical treatment of cardiomyopathy triggered by DNR.
葛根素是从葛根中提取的，具有多种药理作用，包括抗氧化、抗炎、神经保护和心脏保护作用。本研究旨在探讨葛根素对柔红霉素（DNR）诱导的 H9c2 细胞凋亡的影响，并探讨其潜在的作用机制。MTT 法和流式细胞术分别用于评估细胞毒性和细胞凋亡，Western blot 分析用于评估包括 caspase-3、Akt 和磷酸化 Akt（p-Akt）在内的蛋白表达水平的变化。还使用负载 Fura-2 的细胞进行细胞内钙离子（Ca2+）的比率成像。我们的结果表明，葛根素预处理通过抑制细胞凋亡保护 H9c2 细胞免受 DNR 诱导的细胞毒性，这也通过 cleaved caspase-3 表达的降低得到证实。此外，p-Akt 激活与葛根素的抑制作用有关。葛根素预处理后，细胞外 Ca2+内流受到抑制，导致细胞内 Ca2+水平降低；这种作用被 LY294002[磷脂酰肌醇 3-激酶（PI3K）抑制剂]所阻断。Ca2+内流的抑制表明 PI3K/Akt 信号通路参与了葛根素抑制 H9c2 细胞凋亡的作用。综上所述，我们的研究结果表明，葛根素通过抑制 Ca2+内流激活 PI3K/Akt 信号通路，减轻 DNR 诱导的 H9c2 细胞凋亡，提示葛根素可能是一种潜在的心脏保护剂，可用于临床治疗 DNR 引发的心肌病。
