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黄芪和三七活性成分提取物组合对氧化损伤PC12细胞凋亡、活性氧及线粒体膜电位的影响

[Effects of the combination of active component extracts from Astragalus membranaceus and Panax notoginseng on apoptosis, reactive oxygen species and mitochondrial membrane potential of PC12 cells with oxidative injury].

作者信息

Huang Xiao-ping, Liu Xiao-dan, Deng Chang-qing

机构信息

College of Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China.

出版信息

Zhong Xi Yi Jie He Xue Bao. 2012 Oct;10(10):1127-34. doi: 10.3736/jcim20121009.

Abstract

OBJECTIVE

To explore the effects and mechanisms of combining astragaloside IV (the effective component of Astragalus membranaceus) with notoginsenoside R1, ginsenoside Rb1 and ginsenoside Rg1 (the effective components of Panax notoginseng) against oxidative injury in PC12 cells induced by cobalt chloride (CoCl₂).

METHODS

CoCl₂ was used to stimulate PC12 cells to induce injury after transdifferentiation with nerve growth factor. Then the PC12 cells were divided into 10 groups and cultured with corresponding drugs. After culture, apoptotic cells were tested by using Hocchst 33258 fluorescent staining, the level of mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 fluorescent staining and the content of reactive oxygen species (ROS) in PC12 cell was measured by dichlorofluorescin diacetate fluorescent staining.

RESULTS

CoCl₂ induced apoptosis along with the obvious decrease of MMP as well as overproduction of ROS in PC12 cells. Astragaloside IV, ginsenosides Rg1, ginsenosides Rb1 and notoginsenoside R1 had inhibition effects in different degree on PC12 cell apoptosis induced by CoCl₂, reduced the overproduction of ROS and the decrease of MMP. The effects of the combination were better than those of active component alone.

CONCLUSION

Active components extracted from Astragalus and Panax notoginseng can inhibit PC12 cell apoptosis induced by oxidative injury, furthermore, the effects were enhanced by combination of these components, which may be associated with jointly antagonizing the generation of ROS and raising MMP.

摘要

目的

探讨黄芪有效成分黄芪甲苷与三七有效成分三七皂苷R1、人参皂苷Rb1和人参皂苷Rg1联合应用对氯化钴(CoCl₂)诱导的PC12细胞氧化损伤的影响及其机制。

方法

用神经生长因子诱导PC12细胞转分化后,用CoCl₂刺激PC12细胞诱导损伤。然后将PC12细胞分为10组,并用相应药物培养。培养后,采用Hocchst 33258荧光染色检测凋亡细胞,用罗丹明123荧光染色分析线粒体膜电位(MMP)水平,用二氯荧光素二乙酸酯荧光染色测定PC12细胞中活性氧(ROS)含量。

结果

CoCl₂诱导PC12细胞凋亡,同时伴有MMP明显降低以及ROS过量产生。黄芪甲苷、人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1对CoCl₂诱导的PC12细胞凋亡有不同程度的抑制作用,减少了ROS的过量产生和MMP的降低。联合用药的效果优于单一活性成分。

结论

黄芪和三七提取的活性成分可抑制氧化损伤诱导的PC12细胞凋亡,此外,这些成分联合应用可增强其效果,这可能与共同拮抗ROS的产生和提高MMP有关。

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