School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China.
Department of Pharmacy, The First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510405, P.R. China.
Mol Med Rep. 2018 Feb;17(2):3267-3273. doi: 10.3892/mmr.2017.8242. Epub 2017 Dec 11.
Bone marrow mesenchymal stem cell (BMSC) transplants are promising for the treatment of certain central nervous system diseases. However, oxidative stress is one of the major factors that may limit the survival of the transplanted BMSCs. The present study investigated the effect of pretreatment with gigantol on hydrogen peroxide (H2O2)‑induced apoptosis in rat BMSCs (rBMSCs) and the potential underlying mechanisms. The results demonstrated that gigantol pretreatment significantly inhibited H2O2‑induced apoptosis of rBMSCs. rBMSCs were incubated with 600 µM H2O2 in the absence or presence of different doses of gigantol (1‑100 µM). Cell viability and apoptosis ratios were assessed by MTT assays and flow cytometry, respectively. Morphological alterations and reactive oxygen species were measured by the fluorescent‑based methods of Hoechst staining and dichlorodihydrofluorescein diacetate, respectively. Furthermore, the protein levels of phosphorylated‑protein kinase B (Akt), B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2‑associated X (Bax), caspase‑3 and caspase‑9 were investigated by western blotting. Following incubation with H2O2 for 2 h, gigantol significantly inhibited the H2O2‑induced reductions in the cell viability of rBMSCs in a dose‑dependent manner. Furthermore, gigantol upregulated Akt phosphorylation and Bcl‑2 expression, downregulated Bax expression, and reduced the expression of caspase‑3 and caspase‑9 in H2O2‑treated rBMSCs, whereas an opposite effect was detected when LY294002, an inhibitor of phosphatidylinositol 3‑kinase, was administered in combination with gigantol. These results indicate that gigantol may be developed as a promising neuroprotective agent for successful MSC transplantation in ischemic diseases.
骨髓间充质干细胞（BMSC）移植对于治疗某些中枢神经系统疾病具有广阔的前景。然而，氧化应激是可能限制移植 BMSC 存活的主要因素之一。本研究旨在探讨预处理 gigantol 对过氧化氢（H2O2）诱导的大鼠 BMSC（rBMSC）凋亡的影响及其潜在机制。结果表明，gigantol 预处理可显著抑制 H2O2 诱导的 rBMSC 凋亡。将 rBMSC 与 600 μM H2O2 在无 gigantol 或不同浓度 gigantol（1-100 μM）存在的情况下孵育。通过 MTT 检测试剂盒和流式细胞术分别评估细胞活力和凋亡率。通过 Hoechst 染色和二氯荧光素二乙酸酯的荧光法分别测量形态改变和活性氧。此外，通过 Western blot 检测磷酸化蛋白激酶 B（Akt）、B 细胞淋巴瘤-2（Bcl-2）、Bcl-2 相关 X（Bax）、半胱氨酸天冬氨酸蛋白酶-3 和半胱氨酸天冬氨酸蛋白酶-9 的蛋白水平。H2O2 孵育 2 h 后，gigantol 呈剂量依赖性地显著抑制 H2O2 诱导的 rBMSC 细胞活力降低。此外，gigantol 上调 Akt 磷酸化和 Bcl-2 表达，下调 Bax 表达，降低 H2O2 处理的 rBMSC 中 caspase-3 和 caspase-9 的表达，而在用磷脂酰肌醇 3-激酶抑制剂 LY294002 与 gigantol 联合给药时，观察到相反的效果。这些结果表明，gigantol 可能作为一种有前途的神经保护剂，用于成功进行缺血性疾病中的 MSC 移植。
