基于内质网应激的牡荆素抗糖氧剥夺后恢复的细胞损伤作用及机制

    Effect and Mechanism of Vitexin on the Recovery of Cells After Glucose and Oxygen Deprivation Based on Endoplasmic Reticulum Stress

    • 摘要:
      目的  探讨牡荆素通过内质网应激相关通路调控糖氧剥夺后恢复(oxygen glucose deprivation/re-oxygenation,OGD/R) SH-SY5Y细胞及PC12细胞炎症因子,从而阐明牡荆素对OGD/R细胞损伤的保护作用机制。
      方法 建立OGD/R诱导SH-SY5Y细胞及PC12细胞模型,将其分为对照组(Control)、模型组(OGD/R)、OGD/R+0.01 µmol·L−1牡荆素组、OGD/R+ 0.1 µmol·L−1牡荆素组及OGD/R+1 µmol·L−1牡荆素组,CCK-8法检测细胞活力的影响;流式细胞数检测活性氧(reactive oxygen species,ROS)水平;免疫荧光检测髓鞘碱性蛋白(myelin basic protein,MBP);PCR检测内质网应激相关基因GRP78CHOP的mRNA水平及炎症因子IL-6TNF-α的表达水平;Western blotting检测内质网应激相关蛋白GRP78、ATF6、ATF4、CHOP的表达水平。
      结果 1 µmol·L−1牡荆素能显著升高细胞活力(P<0.01),显著降低ROS水平(P<0.01),显著升高MBP蛋白表达水平(P<0.01);抑制内质网应激相关蛋白GRP78、ATF6、ATF4、CHOP表达,降低内质网应激相关基因GRP78CHOP的mRNA水平;降低细胞炎症因子IL-6TNF-α
      结论 在SH-SY5Y细胞及PC12细胞中,一定剂量的牡荆素能有效抑制ROS产生,促进MBP表达,缓解受损神经细胞内质网应激导致的炎症损伤,发挥一定的神经保护作用。

       

      Abstract:
      OBJECTIVE  To investigate the regulation of vitexin on inflammatory factors of oxygen glucose deprivation/re-oxygenation(OGD/R) SH-SY5Y cells and PC12 cells through endoplasmic reticulum stress-related pathways, and to clarify the protective mechanism of vitexin on cells against OGD/R injury.
      METHODS  OGD/R-induced SH-SY5Y cell and PC12 cell models were established. They were divided into control group, model group(OGD/R), OGD/R+0.01 µmol·L−1 vitexin group, OGD/R+0.1 µmol·L−1 vitexin group and OGD/R+1 µmol·L−1 vitexin group. CCK-8 assay was performed to evaluate the impact on cell viability. Flow cytometry was used to measure the level of reactive oxygen species(ROS). Immunofluorescence was used to detect the expression of myelin basic protein(MBP). PCR was performed to measure the mRNA levels of endoplasmic reticulum stress-related genes GRP78 and CHOP, as well as the expression levels of inflammatory factors IL-6 and TNF-α. Western blotting was conducted to detect the expression of endoplasmic reticulum stress-related proteins GRP78, ATF6, ATF4, and CHOP.
      RESULTS 1 µmol·L−1 vitexin significantly increased cell viability(P<0.05), reduced ROS levels(P<0.01), and increased the expression level of MBP protein(P<0.01). It inhibited the expression of endoplasmic reticulum stress-related proteins GRP78, ATF6, ATF4, and CHOP, and decreased the mRNA levels of endoplasmic reticulum stress-related genes GRP78 and CHOP. It also reduced the levels of inflammatory cytokines IL-6 and TNF-α.
      CONCLUSION  In SH-SY5Y cells and PC12 cells, a certain dose of vitexin can effectively inhibite ROS production, promote MBP expression, alleviate the inflammatory injury caused by endoplasmic reticulum stress of damaged nerve cells, and play a certain neuroprotective role.

       

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