Wang Jing-Kang, Wang Li-Chao, Jiang Yong, Tu Peng-Fei, Zeng Ke-Wu
State Key Laboratory of Natural and Biomimetic Drugs,Peking University Beijing 100191,China.
Zhongguo Zhong Yao Za Zhi. 2019 Jul;44(13):2686-2690. doi: 10.19540/j.cnki.cjcmm.20181214.001.
To investigate the inhibitory effects and mechanism of Cistanche tubulosa ethanol extract( CTEE) against oxygen-glucose deprivation/reperfusion( OGD/R)-induced PC12 cells neuronal injury. In this study,OGD/R-induced PC12 cells were used to explore the neuroprotective effects of CTEE( 12. 5,25,50 mg·L-1) by detecting cell viability with MTT assay,apoptosis with AO/EB and Hoechst 33258,mitochondrial membrane potential changes with JC-1 staining,mitochondrial oxidative stress with MitoSOX staining,as well as the apoptosis-related protein expression( PARP,cleaved PARP,caspase-3,cleaved caspase-3,Bax,Bcl-2) with Western blot. RESULTS:: showed that CTEE effectively protected OGD/R-induced neuronal injury and increased the survival rate of PC12 cells.AO/EB and Hoechst 33258 staining showed that CTEE could effectively inhibit apoptosis. Moreover,JC-1 and MitoSOX staining results showed that CTEE decreased mitochondrial stress and mitochondrial membrane potential imbalance in PC12 cells in a concentration-dependent manner. Meanwhile,CTEE could obviously suppress the activation of key proteins in mitochondrial apoptosis pathway such as caspase-3 and PARP,and significantly inhibit the rise of Bax and down-regulation of Bcl-2. In conclusion,CTEE has obvious protective effects on OGD/R-induced PC12 cells neuronal injury,potentially via inhibiting mitochondrial oxidative stress and apoptosis-related signaling pathway.
探讨管花肉苁蓉乙醇提取物(CTEE)对氧糖剥夺/复灌注(OGD/R)诱导的PC12细胞神经损伤的抑制作用及其机制。本研究采用OGD/R诱导的PC12细胞,通过MTT法检测细胞活力、AO/EB和Hoechst 33258检测细胞凋亡、JC-1染色检测线粒体膜电位变化、MitoSOX染色检测线粒体氧化应激以及Western blot检测凋亡相关蛋白表达(PARP、裂解的PARP、caspase-3、裂解的caspase-3、Bax、Bcl-2),探讨CTEE(12.5、25、50 mg·L-1)的神经保护作用。结果显示,CTEE能有效保护OGD/R诱导的神经损伤,提高PC12细胞的存活率。AO/EB和Hoechst 33258染色显示CTEE能有效抑制细胞凋亡。此外,JC-1和MitoSOX染色结果显示CTEE能以浓度依赖的方式降低PC12细胞的线粒体应激和线粒体膜电位失衡。同时,CTEE能明显抑制线粒体凋亡途径中关键蛋白如caspase-3和PARP的激活,并显著抑制Bax的升高和Bcl-2的下调。综上所述,CTEE对OGD/R诱导的PC12细胞神经损伤具有明显的保护作用,其机制可能是通过抑制线粒体氧化应激和凋亡相关信号通路实现的。