远志皂苷元对脂多糖诱发神经细胞炎症损伤的保护作用
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1.昆明医科大学附属延安医院,昆明 650051; 2.中南大学湘雅医学院,长沙 410013; 3.上海中医药大学中药研究所,上海 201203

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R-33

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Senegenin protects against lipopolysaccharide-induced neurite toxicity in a nerve cell model
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1.the Yan An Hospital Affilated Kun Ming University, Kunming 650051, China. 2. Xiangya School of Medicine,Central South University, Changsha 410013. 3. Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203

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    摘要:

    目的 研究具有神经营养作用的远志皂苷元对神经细胞炎症因子的影响,探讨远志皂苷元是否具有降低神经细胞炎症反应的作用,从而为缓解抗肿瘤药物的毒副作用寻找新的突破口。 方法 体外培养小胶质细胞 BV2 细胞,采用脂多糖(lipopolysaccharide,LPS)制备细胞炎症模型,观察不同浓度的远志皂苷元对 LPS 诱导的细 胞炎症因子—一氧化氮( nitric oxide, NO) 的影响,从而发现较佳的远志皂苷元浓度;远志皂苷元 2 μmol / L、2. 5 μmol / L 及 3 μmol / L 和阳性对照组地塞米松 10 μmol / L,主要通过 Griess Reagent 法检测炎症介质 NO 浓度;定量聚合酶链反应(quantitative polymerase chain reaction, QPCR)法检测环氧化酶-2( cyclooxygenase-2, COX-2) mRNA 表达量;Western blot 法检测 COX-2 蛋白表达量; 并用 LPS 诱导后的 BV2 炎症上清刺激 SH-SY5Y 细胞,再进行其细胞 活力和 NO 浓度检测。 结果 不同浓度远志皂苷元对 LPS 诱导的 BV2 细胞产生 NO 的影响不同。 与 LPS 组相比较, (5 μmol / L、25 μmol / L、50 μmol / L、100 μmol / L)剂量组远志皂苷元抗炎作用不明显,且 50 μmol / L 及 100 μmol / L 远志 皂苷元可提高炎症介质 NO 的浓度,具有明显的促炎作用;而(1 μmol / L、2 μmol / L、4 μmol / L、8 μmol / L)剂量组中 发现 1 μmol / L、2 μmol / L 远志皂苷元具有明显抗炎症作用(P<0. 05 或 P<0. 01),综上结果发现低剂量远志皂苷元 具抗炎作用;(1. 5 μmol / L、2 μmol / L、2. 5 μmol / L、3 μmol / L)剂量组远志皂苷元均具有明显的降低 NO 浓度的作 用,但以 2 μmol / L 尤为显著(P<0. 001),据该结果后续实验中远志皂苷元分为低、中、高(2 μmol / L、2. 5 μmol / L、 3 μmol / L)剂量组;各组 COX-2 mRNA 表达情况对比发现,与 LPS 组相比,远志皂苷元 2 μmol / L、2. 5 μmol / L 组中 COX-2 mRNA 表达显著降低(P<0. 001),仍稍弱于地塞米松组;另外相比 LPS 组,远志皂苷元 2 μmol / L、2. 5 μmol / L、3 μmol / L 剂量组均可降低 COX-2 蛋白表达水平,以 2 μmol / L 效果最为显著(P<0. 05)。 结论 远志皂苷元可抑 制 LPS 诱导的细胞炎症因子的释放及表达,表明具有神经营养作用的远志皂苷元可能还具有保护神经细胞炎症损伤的作用。

    Abstract:

    Objective To study the neurotrophic and anti-inflammatory effects of senegenin on inflammatory factors in nerve cells to identify novel drugs that alleviate the toxic and side effects of anti-tumor drugs. Methods Microglial BV2 cells were cultured in vitro, and lipopolysaccharide (LPS) was used to prepare the cell inflammation model. The effects of different concentrations of senegenin (2, 2. 5, 3 μmol / L) on the LPS-induced cellular inflammatory factor nitric oxide (NO) were observed to determine the optimal dose of senegenin compared with the positive control group ( dexamethasone 10 μmol / L). The concentration of NO in inflammatory medium was detected by Griess Reagent. Results The effects of different concentrations of senegenin on the production of NO in LPS-induced BV2 cells were different. Compared with the LPS group, the anti-inflammatory effects of senegenin (5, 25, 50, 100 μmol / L) were not obvious. Moreover, the concentration of NO in the inflammatory medium was increased by 50 and 100, indicating a proinflammatory effect. In addition, in the senegenin 1, 2, 4, and 8 μmol / L dose range, 1, 2, and 2 kept up with each other and had significant anti-inflammatory effects (P < 0. 05 or P < 0. 01). The low doses of senegenin had anti-inflammatory effects. All doses of senegenin, especially 2 μmol / L (P < 0. 001), significantly reduced the concentration of NO. According to this result , senegenin was divided into low, medium and high dose groups in a follow-up test. CoX-2 mRNA expression in the senegenin 2 and 2. 5 μmol / L groups was significantly decreased (P < 0. 001) compared with the LPS group, but was slightly lower than that in the dexamethasone group. In addition, compared with the LPS group, 2, 2. 5, and 3 μmol / L senegenin reduced CoX-2 protein levels, with the most significant effect shown for 2 μmol / L senegenin ( P < 0. 05). Conclusions Senegenin reduces the release and expression of inflammatory factors in microglia induced by LPS, suggesting senegenin has neurotrophic effects that might protect nerve cells from inflammatory injury.

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皮 婷,梁月琴,欧雯励蓉,朱 晗,陶彦林,金醒昉.远志皂苷元对脂多糖诱发神经细胞炎症损伤的保护作用[J].中国比较医学杂志,2020,30(11):52~58.

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  • 收稿日期:2020-04-07
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  • 在线发布日期: 2020-12-25
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