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首页 > 过刊浏览>2022年第32卷第7期 >18-26. DOI:10. 3969 / j.issn.1671-7856. 2022. 07. 003
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连翘提取物通过调节巨噬细胞凋亡和极化抑制 LPS 诱导的炎症作用
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1.中国农业科学院农产品加工研究所,北京 100193;2.西南医科大学附属医院急诊科,四川 泸州 646000; 3.四川省康复医院,成都 611139;4.重庆医科大学附属第一医院神经内科,重庆 400016

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R-33


Forsythia suspensa extract inhibits LPS-induced inflammation by regulating macrophage apoptosis and polarization
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Affiliation:

1.Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China. 2. Department of Emergency Medicine, the Affiliated Hospital of Southwest Medical University, Luzhou 646000. 3. Sichuan Rehabilitation Hospital, Chengdu 611139; 4. Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016

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    摘要:

    目的 研究连翘提取物(FSE)对脂多糖(LPS)诱导炎症的抑制作用,以及 LPS 诱导的 RAW264. 7 细胞极化模型建立。 方法 MTT 法检测 FSE 和 LPS 对 RAW264. 7 细胞的毒性作用;倒置显微镜检测不同浓度的 FSE 对 LPS 刺激后的细胞形态变化,酶标仪检测细胞上清液的一氧化氮(NO)生成量;荧光显微镜检测细胞内活性氧 (ROS)和细胞凋亡的荧光强度;Real-time PCR 法检测巨噬细胞中 iNOS、TNF-α、IL-1β、IL-6、IL-10、Arginase-1 和 CD206 mRNA 的表达水平。 结果 MTT结果发现 FSE 在 50 μg / mL 浓度时无细胞毒性,LPS 作用 36 h 时,不同浓度 FSE 对 LPS 刺激的细胞形态变化和 NO 生成有抑制作用,同时 FSE 可以抑制 LPS 刺激后活性氧(ROS)的减少和细胞的凋亡,Real-time PCR结果显示,用不同浓度(100、250、500、750 ng / mL) LPS 处理细胞 24 h,或者用 500 ng / mL 浓度的 LPS 处理细胞不同时间后,RAW264. 7 细胞 M1 型标记基因(TNF-α、IL-1β、iNOS、IL-6)mRNA 的表达水平明显升高,RAW264. 7 细胞 M2 型标记基因(IL-10、Arginase-1、CD206)mRNA 的表达水平明显降低;FSE 可降低 LPS 诱导的 M1 型标记基因 mRNA 的表达水平,升高 M2 型标记基因 mRNA 的表达水平。 结论 FSE 可通过抑制 RAW264. 7 细胞凋亡和促进 RAW264. 7 细胞的极化来抑制 LPS 诱导的炎症反应。

    Abstract:

    Objective We investigated the inhibitory effect of Forsythia suspensa extract ( FSE ) on lipopolysaccharide ( LPS)-induced inflammation and establishment of an LPS-induced polarization model of RAW264. 7 cells. Methods MTT assays were used to assess cytotoxic effects of FSE and LPS on RAW264. 7 cells. Inverted microscopy was used to assess morphological changes of cells after stimulation with LPS and various concentrations of FSE. An enzyme marker was used to detect nitric oxide (NO) in culture supernatants. Fluorescence microscopy was used to detect intracellular reactive oxygen species (ROS) and apoptosis. ; Real-time PCR was used to detect iNOS expression in macrophages. iNOS, TNF-α, IL-1β, IL-6, IL-10, Arginase-1 and CD206 mRNA expression was measured by Real-time PCR. Results FSE was not cytotoxic at 50 μg / mL. At 36 h of LPS stimulation, various concentrations of FSE inhibited LPS-induced morphological changes, NO production and FSE inhibited LPS-induced the reduction of reactive oxygen species (ROS) and apoptosis. Expression of M1-type marker genes (TNF-α, IL-1β, iNOS and IL-6) in RAW264. 7 cells was significantly increased after treatment with various concentrations (100, 250, 500 and 750 ng / mL) of LPS for 24 h or with 500 ng / mL LPS for various times. The mRNA levels of M2 marker genes ( IL-10, Arginase-1 and CD206) were significantly decreased in RAW264. 7 cells. FSE decreased the expression of LPS-induced M1 marker genes and increased expression of M2 marker genes. Conclusions FSE inhibits LPS-induced inflammatory responses by inhibiting RAW264. 7 cell apoptosis and promoting polarization.

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罗福龙,李文举,吴小会,钟 武,范 蓓,贯东艳,王凤忠,王 琼.连翘提取物通过调节巨噬细胞凋亡和极化抑制 LPS 诱导的炎症作用[J].中国比较医学杂志,2022,32(7):18~26.

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  • 收稿日期:2021-11-15
  • 在线发布日期: 2022-11-09
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