3,4-二羟基苯乙酮通过抗炎抗氧化作用缓解脂多糖诱导的急性肺损伤

doi:10.5246/jcps.2021.12.082

摘要/Abstract

摘要：

炎症反应和氧化应激的增强会导致急性肺损伤(ALI), 控制炎症和氧化可改善ALI。本研究的目的是确定3,4-二羟基苯乙酮(1)是否通过抑制炎症和氧化来改善脂多糖(LPS)诱导的ALI。在本研究中, 化合物1降低了LPS诱导的RAW 264.7细胞炎性细胞因子和氧化应激。此外, 化合物1抑制LPS诱导RAW 264.7细胞p65磷酸化和p65核转位, 增加抗氧化蛋白核因子红细胞2相关因子2(Nrf-2)和血红素氧合酶-1(HO-1)的表达, 并能改善LPS诱导的ALI, 减少炎症细胞因子和抗氧化蛋白的表达, 以及核因子κB(NF-κB)信号通路激活。化合物1具有抗炎、抗氧化作用, 有望成为治疗ALI的有效药物。

关键词: 3,4-二羟基苯乙酮, 急性肺损伤, 抗炎, 抗氧化

Abstract:

Enhanced inflammatory response and oxidative stress cause acute lung injury (ALI). Controlling inflammation and oxidation can ameliorate ALI. In the present study, we aimed to determine whether 3,4-Dihydroxyacetophenone (compound 1) could ameliorate lipopolysaccharide (LPS)-induced ALI by suppressing inflammation and oxidation. In this study, compound 1 reduced LPS-induced inflammatory cytokines and oxidative stress in RAW 264.7 cells. Moreover, compound 1 suppressed the expression of inflammatory protein p65, inhibited IkBα phosphorylation, decreased the nuclear translocation of p65, and increased the expressions of anti-oxidative protein nuclear factor erythroid 2-related factor 2 (Nrf-2) and heme oxygenase-1 (HO-1), which was reduced by LPS, in leukemia cells in mouse macrophage (RAW 264.7) cells. Furthermore, compound 1 could also ameliorate LPS-induced ALI in vivo, with a reduction of inflammatory cytokines, oxidative stress, and nuclear factor-kappa B (NF-κB) signaling pathway activation. This study emphasized the anti-inflammatory and anti-oxidative activities of compound 1, which could be a valuable therapeutic agent against ALI.

Key words: 3,4-Dihydroxyacetophenone, Acute lung injury, Anti-inflammatory, Antioxidant

Supporting:

徐银燕, 朱敏, 周开珩, 宋涛涛, 黄丽丽. 3,4-二羟基苯乙酮通过抗炎抗氧化作用缓解脂多糖诱导的急性肺损伤[J]. 中国药学(英文版), 2021, 30(12): 956-968.

Yinyan Xu, Min Zhu, Kaiheng Zhou, Taotao Song, Lili Huang. 3,4-Dihydroxyacetophenone alleviates lipopolysaccharide-induced acute lung injury as a potential anti-inflammatory and anti-oxidative agent[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(12): 956-968.

图/表 6

Figure 1. Chemical structure of compound 1.

Figure 2. Compound 1 suppresses the LPS-induced inflammation and oxidative damage in RAW 264.7 cells. Compound 1 decreased the expressions of TNF-α (A), IL-1β (B), IL-2 (C), IL-6 (D), and IL-12 (E). GAPDH was used as an internal control. (F) Compound 1 decreased ROS levels in LPS-treated RAW 264.7 cells. ###P < 0.001 significantly different from the control group. ***P < 0.001, **P < 0.01, *P < 0.05 significantly different from the LPS group.

Figure 3. Effects of compound 1 on the NF-κB and Nrf-2/HO-1 signaling pathways in vitro. (A) The nuclear translocation of p65 was detected using immunofluorescence staining. (B) Compound 1 regulated the expressions of Nrf-2, HO-1, p-p65 and p-IκBα at the protein level. (C) The relative protein expression was calculated as the fold change of the control group. The results were presented as means ± SEM (n = 3–6). *P < 0.05, **P < 0.01.

Figure 4. Protective effects of compound 1 on LPS-induced ALI in mice. (A) Lung histopathological analysis was performed using hematoxylin and eosin staining to determine the protective effect on LPS-induced pathological damage. (B) The lung wet/dry weight ratio was calculated. (C) The total number of cells in BALF was calculated. (D) The protein concentrations in BALF were measured using a BCA protein assay kit. Control: control group; LPS: LPS group; 1: compound 1 alone group; 1 + LPS: compound 1 + LPS group. The results were presented as means ± SEM (n = 3–6). *P < 0.05, **P < 0.01, ***P < 0.001 vs. the LPS group.

Figure 5. Compound 1 inhibits LPS-induced secretion of IL-1β, IL-6, and TNF-α and oxidative damage in mice. The levels of TNF-α (A), IL-6 (B), and IL-1β (C) in BALF were measured by ELISA. The MDA content (D) and SOD activity (E) were determined as per the manufacturer’s protocol. Control: control group; LPS: LPS group; 1: compound 1 alone group; 1 + LPS: compound 1 + LPS group. Results were presented as means ± SEM (n = 3–6). *P < 0.05, **P < 0.01, ***P < 0.001 vs. the LPS group.

Figure 6. The NF-κB pathway is suppressed by compound 1 in the LPS-induced mice model. (A) Immunofluorescence staining with a p65 antibody was performed to detect the nuclear translocation of p65. (B) The levels of p-p65 and p-IκBα in lung tissues were measured by Western blotting analysis. (C) The relative protein expression was calculated as the fold change of the control group. The results were presented as means ± SEM (n = 3–6). **P < 0.01, ***P < 0.001.

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