Department of Orthopedics, The Second Hospital of Jilin University, Changchun, Jilin, China (mainland).
Central Laboratory, The Second Hospital of Jilin University, Changchun, Jilin, China (mainland).
Med Sci Monit. 2018 May 14;24:3159-3167. doi: 10.12659/MSM.910718.
BACKGROUND The oxidative stress environment of pathological tissue has an adverse effect on the survival of bone marrow mesenchymal stem cells (BMSCs) transplantation. Ginkgo biloba L. extract (EGB) has a potent antioxidant effect. In this research, we assessed the protective effects of EGB and EGB-Containing Serum (EGB CS) on BMSCs against injury induced by hydrogen peroxide (H2O2). MATERIAL AND METHODS BMSCs were pretreated with EGB or EGB CS and treated with H2O2. The cell counting kit-8 (CCK-8) method was utilized to detect cell viability. The DCFH-DA Fluorescent Kit method was used to detect intracellular ROS level. Malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and (CAT) were determined. The Hoechst staining assay and qRT-PCR assay were utilized to evaluate the effect of EGB on cell apoptosis. Mitogen-activated protein kinases (MAPKs) signaling pathway were detected by western blot analysis. RESULTS Compared to the H2O2 group, the number of apoptotic cells in the EGB and EGB CS pretreated groups significantly decreased. The mRNA expression ratio of Bax/Bcl-2 was also decreased. EGB and EGB CS can reduce the production of ROS in BMSCs exposed to H2O2. SOD, GSH-Px and CAT activities were significantly higher compared with those with H2O2 group. Furthermore, EGB or EGB CS pretreatment decreased the protein levels of p-p38MAPK and p-JNK in BMSCs compared to the H2O2 group. CONCLUSIONS Our findings suggested that EGB and EGB CS have protective effect on BMSCs against oxidative stress injury and increase the survival rate of BMSCs transplantation by regulating p38MAPK and JNK signaling.
病理性组织的氧化应激环境对骨髓间充质干细胞（BMSCs）移植的存活有不利影响。银杏叶提取物（EGB）具有很强的抗氧化作用。本研究评估了 EGB 和含 EGB 血清（EGB CS）对双氧水（H2O2）诱导的 BMSCs 损伤的保护作用。
用 EGB 或 EGB CS 预处理 BMSCs，然后用 H2O2 处理。用细胞计数试剂盒-8（CCK-8）法检测细胞活力。用二氯荧光素二乙酸酯（DCFH-DA）荧光试剂盒法检测细胞内 ROS 水平。测定丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和（CAT）。用 Hoechst 染色法和 qRT-PCR 法评估 EGB 对细胞凋亡的影响。用 Western blot 分析检测丝裂原活化蛋白激酶（MAPKs）信号通路。
与 H2O2 组相比，EGB 和 EGB CS 预处理组凋亡细胞数量明显减少。Bax/Bcl-2 的 mRNA 表达比值也降低。EGB 和 EGB CS 可减少暴露于 H2O2 的 BMSCs 中 ROS 的产生。与 H2O2 组相比，SOD、GSH-Px 和 CAT 活性显著升高。此外，与 H2O2 组相比，EGB 或 EGB CS 预处理可降低 BMSCs 中 p-p38MAPK 和 p-JNK 的蛋白水平。
我们的研究结果表明，EGB 和 EGB CS 对 BMSCs 氧化应激损伤具有保护作用，并通过调节 p38MAPK 和 JNK 信号通路增加 BMSCs 移植的存活率。
