Zheng Long, Lu Xuli, Yang Shengtao, Zou Ying, Zeng Fanke, Xiong Shaohao, Cao Yupo, Zhou Wei
Key Laboratory of Tropical Crop Products Processing of Ministry of Agriculture and Rural Affairs, Agricultural Products Processing Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang, Guangdong, China.
Front Nutr. 2023 Mar 9;10:1093036. doi: 10.3389/fnut.2023.1093036. eCollection 2023.
Gamma-aminobutyric acid (GABA), one of the main active components in leaves, can be widely used to treat multiple diseases including inflammation.
In this study, the anti-inflammatory activity and the underlying anti-inflammatory mechanism of the GABA-enriched leaves fermentation broth (MLFB) were investigated on lipopolysaccharide (LPS)-induced RAW 264.7 cells model. The key active components changes like total flavonoids, total polyphenols and organic acid in the fermentation broth after fermentation was also analyzed.
ELISA, RT-qPCR and Western blot results indicated that MLFB could dose-dependently inhibit the secretions and intracellular expression levels of pro-inflammatory cytokines like 1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). Furthermore, MLFB also suppressed the expressions of prostaglandin E (PGE) and inducible nitric oxide synthase (iNOS). Moreover, the mRNA expressions of the key molecules like Toll-like receptor 4 (TLR-4) and nuclear factor (NF)-κB in the NF-κB signaling pathway were also restrained by MLFB in a dose-dependent manner. Besides, the key active components analysis result showed that the GABA, total polyphenols, and most organic acids like pyruvic acid, lactic acid as well as acetic acid were increased obviously after fermentation. The total flavonoids content in MLFB was still remained to be 32 mg/L though a downtrend was presented after fermentation.
Our results indicated that the MLFB could effectively alleviate LPS-induced inflammatory response by inhibiting the secretions of pro-inflammatory cytokines and its underlying mechanism might be associated with the inhibition of TLR-4/NF-κB inflammatory signaling pathway activation. The anti-inflammatory activity of MLFB might related to the relative high contents of GABA as well as other active constituents such as flavonoids, phenolics and organic acids in MLFB. Our study provides the theoretical basis for applying GABA-enriched leaves as a functional food ingredient in the precaution and treatment of chronic inflammatory diseases.
γ-氨基丁酸(GABA)是树叶中的主要活性成分之一,可广泛用于治疗包括炎症在内的多种疾病。
在本研究中,以脂多糖(LPS)诱导的RAW 264.7细胞模型,研究了富含GABA的树叶发酵液(MLFB)的抗炎活性及其潜在的抗炎机制。还分析了发酵后发酵液中总黄酮、总多酚和有机酸等关键活性成分的变化。
酶联免疫吸附测定(ELISA)、逆转录定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹法(Western blot)结果表明,MLFB可剂量依赖性地抑制促炎细胞因子如白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)和肿瘤坏死因子-α(TNF-α)的分泌及细胞内表达水平。此外,MLFB还抑制了前列腺素E(PGE)和诱导型一氧化氮合酶(iNOS)的表达。而且,MLFB还以剂量依赖性方式抑制了核因子κB(NF-κB)信号通路中关键分子如Toll样受体4(TLR-4)和核因子(NF)-κB的mRNA表达。此外,关键活性成分分析结果表明,发酵后GABA、总多酚以及大多数有机酸如丙酮酸、乳酸和乙酸明显增加。尽管发酵后总黄酮含量呈下降趋势,但MLFB中的总黄酮含量仍保持在32 mg/L。
我们的结果表明,MLFB可通过抑制促炎细胞因子的分泌有效减轻LPS诱导的炎症反应,其潜在机制可能与抑制TLR-4/NF-κB炎症信号通路激活有关。MLFB的抗炎活性可能与其相对较高含量的GABA以及其他活性成分如黄酮类、酚类和有机酸有关。我们的研究为将富含GABA的树叶作为功能性食品成分应用于慢性炎症性疾病的预防和治疗提供了理论依据。