基于细胞和果蝇模型的狗枣猕猴桃黄酮提取物抗炎和助眠功效研究

Study on Anti-Inflammatory and Sleep-Improving Efficacy of Actinidia kolomikta Fruit Flavonoid Extracts Based on Cellular and Drosophila Models

摘要:

为评价狗枣猕猴桃果实黄酮提取物(Actinidia kolomikta fruit flavonoid extract,AFFE)抗炎、助眠的功效,以巨噬细胞RAW264.7炎症模型、小鼠海马神经元细胞HT22和黑腹雄蝇为实验对象,分析了AFFE对炎症介质、神经递质及睡眠时间的影响。结果表明,对于脂多糖诱导的RAW264.7细胞,250μg/mL AFFE使得其NO、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)生成量分别减少62.71%、47.38%和65.53%,同时TNF-α和IL-6的mRNA表达极显著下调(P<0.01),AFFE表现出良好的抗炎活性。对于HT22细胞,250μg/mL AFFE 能极显著提高神经递质γ-氨基丁酸(γ-aminobutyric acid, GABA)和5-羟色胺(5-hydroxytryptamine,5-HT)产生量(P<0.01),上调GABRA1、GABRA2、5-HT2A受体mRNA表达。在雄蝇睡眠剥夺模型中,雄蝇经质量分数10% AFFE培养基中培养后,全天睡眠时间延长11.58%,与空白对照组无显著性差异。经LC-MS分析,AFFE中共检测出33种活性成分,其中有14种黄酮类物质。研究结果表明,AFFE对神经递质GABA和5-HT生成增长率与炎症介质NO、TNF-α和IL-6分泌抑制率之间有正相关关系,AFFE的助眠作用与其黄酮类物质的抗炎能力有关。

Abstract:

To evaluate the anti-inflammatory and sleep-improving properties of Actinidia kolomikta fruit flavonoid extract (AFFE), macrophage RAW264.7 inflammatory model, mouse hippocampal neuronal cells HT22, and Drosophila melanogaster were used as experimental subjects and the effects of AFFE on inflammatory mediators, neurotransmitters, and sleep duration were analyzed. The results showed that in lipopolysaccharide-induced RAW264.7 cells, 250μg/mL AFFE reduced the production of NO, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) by 62.71%, 47.38%, and 65.53%, respectively. Additionally, the mRNA expressions of TNF-α and IL-6 were significantly down regulated (P<0.01), indicating that AFFE exhibited potent anti-inflammatory activity. In HT22 cells, 250μg/mL AFFE significantly increased the production of the neurotransmitters γ-aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT) (P<0.01) and upregulated the mRNA expression of GABRA1, GABRA2, and 5-HT2A receptors. In the Drosophila melanogaster sleep deprivation model, male flies cultured in a medium containing 10% AFFE showed an 11.58% increase in total sleep duration, with no significant difference compared to the control group. LC-MS analysis identified 33 active components in AFFE, including 14 flavonoids. The study revealed a positive correlation between the increase in GABA and 5-HT production and the inhibition of NO, TNF-α, and IL-6 secretion, suggesting that the sleep-improving effects of AFFE were related to the anti-inflammatory properties of its flavonoid constituents.