摘要
目的探讨绿原酸(CGA)对β淀粉样蛋白25-35(Aβ_(25-35))诱导的PC12细胞损伤的保护作用及其机制。方法采用Aβ_(25-35)诱导PC12细胞构建细胞损伤模型,并用3个不同浓度的CGA分组干预。CCK-8法检测细胞存活率;流式细胞术检测细胞凋亡率;比色法检测细胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活力;Western blot检测细胞核转录因子κB(NF-κB)和白细胞介素1β(IL-1β)表达;ELISA法检测细胞肿瘤坏死因子α(TNF-α)浓度。结果与Control组比较,Model组细胞存活率下降、凋亡率升高,SOD和GSH-Px活力降低、MDA水平升高,NF-κB、IL-1β和TNF-α表达水平升高(P<0.05)。与Model组比较,CGA各组细胞存活率升高、凋亡率下降,SOD和GSH-Px活力升高、MDA水平降低,NF-κB、IL-1β和TNF-α蛋白表达水平下降(P<0.05)。结论 CGA对Aβ_(25-35)致PC12细胞损伤具有保护作用。其机制可能与CGA提高细胞抗氧化能力,抑制NF-κB介导的炎症通路有关。
Objective To observe the protective effect of chlorogenic acid（ CGA） on the death of rat pheochromocytoma（ PC12） cells induced by β-amyloid protein 25-35（ Aβ25-35,and explore its possible mechanisms. Methods Aβ25-35was used to induce PC12 cell death. Cells were interfered with three different concentrations of CGA. CCK-8 kit was used to detect cell viability. Cell apoptosis rate was detected by flow cytometry.Photocolorimetric method was applied to detect the MDA content and GSH-Px and SOD activity. Western blot assay was used to detect the intracellular expression of nuclear factor kappa B（ NF-κB） and interleukin-1β（ IL-1β）. The intracellular concentration of tumor necrosis factor-α（ TNF-α） was measured by enzyme immunoassay assay（ ELISA）. Results Compared with the control group,PC12 cells viability of the model group were significantly reduced. The apoptosis rate of the model group was increased. The levels of intracellular SOD and GSH-Px were decreased and the MDA level was increased. The expressions of NF-κB,IL-1β and TNF-α were increased. Compared with the model group,the PC12 cells viability of CGA groups were significantly increased. The apoptosis rate of CGA groups were reduced. The levels of intracellular SOD and GSH-Px were increased and the MDA level was decreased. The expressions of NF-κB,IL-1β and TNF-α were declined.Conclusion CGA has a protective effect on Aβ25-35 induced PC12 cells death. The mechanisms may be related to the improvement of the antioxidant ability and the inhibition of NF-κB signaling pathway activation.
出处
《遵义医学院学报》
2017年第2期161-165,共5页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:81560562)
贵州省科技厅资助项目(NO:黔科合J字[2013]2314)
贵州省教育厅资助项目(NO:黔教科[2009]0108)
珠海市中药基础及应用研究重点实验室开放课题(NO:ZYKL-2016K6)
