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两种新型脱氧化N-乙酰葡糖胺在脂多糖激活的小鼠巨噬细胞RAW264.7中的抗炎活性

Anti-inflammatory activities of two new deoxygenated N-acetyl glucosamines in lipopolysaccharide-activated mouse macrophage RAW264.7 cells.

作者信息

Le Quang, Zhang Zhichang, Sun Daniel, Cui Quanjun, Yang Xinlin, Hassan Ameer E

机构信息

Dept of Orthopaedic Surgery, University of Virginia, 22903, USA.

Dept of Orthopaedic Surgery, The First Affiliated Hospital of Xinxiang Medical University, Wehui, 453100, Henan, China.

出版信息

Heliyon. 2023 Apr 25;9(5):e15769. doi: 10.1016/j.heliyon.2023.e15769. eCollection 2023 May.

Abstract

BACKGROUND

Glucosamine and N-acetyl-glucosamine (NAG) are amino sugars found in human extracellular matrix with previously described anti-inflammatory effects. Despite mixed results from clinical studies, these molecules have been used extensively in supplements.

OBJECTIVE

We investigated the anti-inflammatory properties of two synthesized derivatives of N-acetyl-glucosamine (NAG), bi-deoxy-N-acetyl-glucosamine (BNAG) 1 and 2.

METHODS

Using mouse macrophage RAW 264.7 cells with lipopolysaccharide (LPS) to induce inflammation, the effects of NAG, BNAG 1, and BNAG 2 on the expression of IL-6, IL-1β, inducible nitric oxide synthase (iNOS) and COX-2 were studied using ELISA, Western blot and quantitative RT-PCR. Cell toxicity and nitric oxide (NO) production were evaluated using WST-1 assay and the Griess reagent, respectively.

RESULTS

Among the three tested compounds, BNAG1 shows the highest inhibition of iNOS, IL-6, TNF α and IL-1β expression and NO production. All three tested compounds show slight inhibition on cell proliferation of RAW 264.7 cells, except that BNAG1 displays a remarkable toxicity at the tested maximum dose of 5 mM.

CONCLUSION

BNAG 1 and 2 exhibit notable anti-inflammatory effects compared to the parent NAG molecule.

摘要

背景

氨基葡萄糖和N-乙酰氨基葡萄糖(NAG)是存在于人体细胞外基质中的氨基糖,具有先前所述的抗炎作用。尽管临床研究结果不一,但这些分子已在补充剂中广泛使用。

目的

我们研究了N-乙酰氨基葡萄糖(NAG)的两种合成衍生物双脱氧-N-乙酰氨基葡萄糖(BNAG)1和2的抗炎特性。

方法

使用脂多糖(LPS)诱导炎症的小鼠巨噬细胞RAW 264.7细胞,采用ELISA、蛋白质免疫印迹法和定量逆转录聚合酶链反应研究NAG、BNAG 1和BNAG 2对白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2)表达的影响。分别使用WST-1法和格里斯试剂评估细胞毒性和一氧化氮(NO)生成。

结果

在三种测试化合物中,BNAG1对iNOS、IL-6、肿瘤坏死因子α(TNFα)和IL-1β表达及NO生成的抑制作用最强。除BNAG1在5 mM的测试最大剂量下显示出显著毒性外,所有三种测试化合物对RAW 264.7细胞的增殖均有轻微抑制作用。

结论

与母体NAG分子相比,BNAG 1和2表现出显著的抗炎作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b748/10163627/ef801551ac2a/gr1.jpg

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