奎宁酸是一种小米来源的类黄酮类物质,为探究奎宁酸的神经保护作用,该研究建立了H2O2诱导的SH-SY5Y细胞氧化应激模型,通过MTT法测定奎宁酸对SH-SY5Y细胞活性影响,同时测定了抗氧化酶超氧化物歧化酶(superoxide dismutase,SOD)的活性和氧化应激标志分子谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)的含量,利用实时荧光定量PCR检测了抗氧化酶和炎症因子表达情况,并用Western blot测定了炎症因子的蛋白表达量。研究结果表明,100 μg/mL的奎宁酸处理显著抑制H2O2引起的SH-SY5Y细胞存活率下降;对细胞抗氧化指标的检测显示,奎宁酸处理可以增强细胞SOD的活性,增加GSH的水平并降低细胞的MDA水平,提高SOD、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)等抗氧化酶基因的表达水平,降低氧化损伤;另外,奎宁酸还可以降低H2O2诱导的SH-SY5Y细胞中炎症因子白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的表达水平,减轻SH-SY5Y细胞的损伤。综上,奎宁酸可以通过抑制细胞氧化应激水平、发挥抗炎作用保护神经细胞不被损伤,为进一步探究小米多酚在氧化应激相关疾病中的作用奠定了基础。
Quinic acid is a flavonoid of millet polyphenols.In this study, the H2O2-induced SH-SY5Y oxidative stress cell model was employed to explore the neuroprotective effect of quinic acid.The effect of quinic acid on the viability of SH-SY5Y cells was measured by the MTT method, and in the meanwhile, the activity of the antioxidant enzyme superoxide dismutase (SOD) and the level of the markers of oxidative stress, glutathione (GSH) and malondialdehyde (MDA), were measured.The expression of antioxidant enzymes and inflammatory factors was detected by real-time fluorescent quantitative PCR, and the protein level of inflammatory factors was determined by Western blot.Results showed that pre-treatment of quinic acid (100 μg/mL) on SH-SY5Y cells could significantly inhibit the decrease of cell survival rate induced by H2O2.The examination of antioxidant indices suggested that quinic acid treatment could upregulate the activity of SOD, increase the level of GSH, reduce that of MDA in SH-SY5Y cells, and upregulate the expression level of antioxidant factors, such as SOD, catalase (CAT), and glutathione peroxidase (GPx), to reduce oxidative damage.In addition, quinic acid could suppress the expression of inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in SH-SY5Y cells induced by H2O2 to alleviate the damage of cells.In a word, quinic acid can protect neural cells from damage by inhibiting oxidative stress and exerting anti-inflammatory function.This study will contribute to the further exploration of the role of millet polyphenols in the prevention and treatment of oxidative stress-related diseases.
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