动物营养学报 >

2018 , Vol. 30 >Issue 9: 3702 - 3709

DOI: https://doi.org/10.3969/j.issn.1006-267x.2018.09.041

饲料营养 Feed science and technology

沙葱总黄酮对脂多糖诱导的小鼠腹腔巨噬细胞炎症介质的影响

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  • 内蒙古农业大学动物科学学院, 呼和浩特 010018
王特日格乐(1994-),男,内蒙古通辽人,硕士研究生,从事动物营养与饲料研究。E-mail:843401225@qq.com

收稿日期: 2018-03-12

  网络出版日期: 2018-09-20

基金资助

国家自然科学基金项目（31260558）

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Effects of Total Flavonoids from Allium mongolicum Regel on Inflammatory Mediators induced by Lipopolysaccharide of Mouse Peritoneal Macrophages

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  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2018-03-12

  Online published: 2018-09-20

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摘要

本试验以脂多糖（LPS）诱导的小鼠腹腔巨噬细胞为炎症模型，旨在研究沙葱总黄酮的抗炎作用。应用CCK-8法筛选沙葱总黄酮对小鼠腹腔巨噬细胞活力具有促进作用的浓度，在此基础上设置对照组、LPS组（应激模型，1 μg/mL LPS）、沙葱总黄酮低剂量组（25 μg/mL沙葱总黄酮+1 μg/mL LPS）、沙葱总黄酮中剂量组（50 μg/mL沙葱总黄酮+1 μg/mL LPS）、沙葱总黄酮高剂量组（100 μg/mL沙葱总黄酮+1 μg/mL LPS）。用Griess法测定细胞上清液中一氧化氮（NO）含量；用酶联免疫吸附试验（ELISA）法测定细胞上清液中肿瘤坏死因子-α（TNF-α）、白介素（IL）-6、IL-1β、IL-10含量；用逆转录聚合酶链式反应（RT-PCR）法测定细胞中TNF-α、IL-6、IL-1β、IL-10、一氧化氮合酶（iNOS）mRNA表达量。结果表明：1）与对照组相比，沙葱总黄酮的添加浓度在25.0~100.0 μg/mL时均能显著提高细胞增殖率（P<0.05）；2）与对照组相比，LPS组能显著提高细胞上清液NO的含量（P<0.05）；与LPS组相比，不同浓度沙葱总黄酮均能显著抑制NO的产生（P<0.05）；3）与对照组相比，LPS组的细胞上清液中TNF-α、IL-6、IL-1β、IL-10含量以及细胞中TNF-α、IL-6、IL-1β、IL-10、iNOS mRNA表达量均显著提高（P<0.05）；与LPS组相比，除了低剂量组IL-1β外，25、50、100 μg/mL沙葱总黄酮浓度显著降低细胞上清液中TNF-α、IL-6、IL-1β含量（P<0.05），除了低剂量组IL-10外，显著增加IL-10含量（P<0.05）；除了低剂量组iNOS外，25、50、100 μg/mL沙葱总黄酮浓度显著抑制细胞中TNF-α、IL-6、IL-1β、iNOS mRNA的表达（P<0.05），有促进IL-10 mRNA表达的趋势，但差异不显著（P>0.05）。综上，沙葱总黄酮对LPS诱导的小鼠腹腔巨噬细胞具有显著的抗炎作用。

关键词： 沙葱; 总黄酮; 脂多糖; 小鼠; 腹腔巨噬细胞; 应激模型

本文引用格式

王特日格乐, 王翠芳, 丹妮, 萨茹丽, 杜红喜, 郭春利, 哈斯额尔敦, 曹琪娜, 敖长金 . 沙葱总黄酮对脂多糖诱导的小鼠腹腔巨噬细胞炎症介质的影响[J]. 动物营养学报, 2018 , 30(9) : 3702 -3709 . DOI: 10.3969/j.issn.1006-267x.2018.09.041

Abstract

In the present study, mouse peritoneal macrophages (MPM) induced by lipopolysaccharide (LPS) were used as an inflammation model to investigate the anti-inflammatory effect of total flavonoids from Allium mongolicum Regel. The CCK-8 method was used to select the concentrations of total flavonoids that could stimulate cell viability of MPM. Based on the concentrations, the cells were treated in control group, LPS group (stress model, 1 μg/mL LPS), total flavonoids low dosage group (25 μg/mL total flavonoids+1 μg/mL LPS), medium dosage group (50 μg/mL total flavonoids+1 μg/mL LPS) and high dosage group (100 μg/mL total flavonoids+1 μg/mL LPS). The Griess method was applied to examine the nitric oxide (NO) content in cell supernatant; the contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β and IL-10 in cell supernatant were examined by the method of ELISA; the mRNA expression levels of TNF-α, IL-6, IL-1β, IL-10 and nitric oxide synthase (iNOS) of cells were tested by reverse transcription and polymerase chain reaction (RT-PCR). The results showed as follows:1) compared with the control, the cell proliferation ratio was significantly increased when the concentration range of total flavonoids was 25.0 to 200.0 μg/mL (P<0.05). 2) Compared with the control, the content of NO in cell supernatant was significantly increased in LPS group (P<0.05); compared with LPS group, different concentrations of total flavonoids could inhibit NO production (P<0.05). 3) Compared with the control, the contents of TNF-α,IL-6,IL-1β,IL-10 in cell supernatant and mRNA expression levels of TNF-α, IL-6, IL-1β, IL-10, iNOS in cells were significantly increased in LPS group (P<0.05); compared with LPS group, when cells were treated with total flavonoids at concoctions of 25, 50 and 100 μg/mL, except for IL-1β in low dosage group, the contents of TNF-α, IL-6 and IL-1β in cell supernatant were significantly decreased (P<0.05), and IL-10 content was significantly increased (P<0.05); except for iNOS in high dosage group, total flavonoids at concoctions of 25, 50 and 100 μg/mL significantly inhibited the mRNA expression of TNF-α,IL-6,IL-1β,iNOS in cells (P<0.05), and trend to improve IL-10 mRNA expression, but the difference was not significant (P>0.05). In conclusion, total flavonoids from Allium mongolicum Regel have a significant anti-inflammatory effect on MPM induced by LPS.

Key words： Allium mongolicum Regel; total flavonoids; LPS; mouse; peritoneal macrophages; stress model

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