发热是最常见的临床疾病之一,其特征是热原感染,恶性肿瘤,炎症,和组织损伤,在其他人中。鞣花酸(EA)通过阻断核因子κB(NF-κB)信号通路,抑制该通路相关蛋白的表达,抑制促炎因子白细胞介素-1β(IL-1β)的水平,白细胞介素-6(IL-6),和肿瘤坏死因子-α(TNF-α),提高抗炎因子IL-10水平,有效缓解炎症症状。此外,EA还可以降低体内丙二醛(MDA)和一氧化氮(NO)的水平,增加超氧化物歧化酶(SOD)的活性,谷胱甘肽(GSH),和过氧化氢酶(CAT),清除氧化自由基,抑制脂质氧化,并实现解热和抗炎作用。这项研究的目的是建立EA和各种炎症标志物之间的关系,如TNF-α,IL-6,IL-1β,前列腺素E2(PGE2),和环磷酸腺苷(cAMP),阐明环氧化酶-2(COX-2)/NF-κB信号通路的作用机制。结合代谢组学分析,我们的研究揭示了EA对多种内源性生物标志物的影响,反映了多组分的特点,多目标,和多途径机制。与脂多糖(LPS)处理的动物相比,随后给药EA显着降低了LPS诱导的直肠温度升高(p<0.05或p<0.01),血清SOD和GSH水平显着升高(p<0.05或p<0.01),显著降低血清MDA、IL-1β,IL-6和TNF-α水平(p<0.05或p<0.01)。此外,与LPS处理的动物相比,随后施用EA显着降低脑脊液cAMP和PGE2水平(p<0.05或p<0.01),cAMP显著降低,5-HT水平显着增加(p<0.05或p<0.01),并显着下调下丘脑中p-NF-κBp65和COX-2蛋白的水平。随后的气相色谱-质谱(GC-MS)代谢物分析显示,在LPS处理4h后分离的血清中检测到12种差异代谢物,在LPS处理后7h收集的血清中检测到10种差异代谢产物。接下来,Pearson相关性分析用于系统表征鉴定的代谢物与TNF-α之间的关系。IL-6,MDA,SOD,PGE2和cAMP。丙酸的水平,吡啶,L-缬氨酸被EA上调,抑制MDA的表达,IL-1β,和TNF-α并增加GSH的活性。肌醇的水平,尿素,2-单半乳糖苷被EA下调,抑制MDA的表达,IL-1β,和TNF-α,增加了SOD和GSH的活性,减少炎症反应,并缓解了氧化应激状态。结合代谢途径分析的结果,我们认为半乳糖代谢的途径,酮体的合成和降解,以及抗坏血酸和醛酸的代谢与EA的解热和抗炎作用密切相关。我们的研究建立了EA和各种炎症标志物之间的关系,如TNF-α,IL-6,IL-1β,PGE2和cAMP,阐明了COX-2/NF-κB信号通路的作用机制。结合代谢组学分析,我们的研究揭示了EA对多种内源性生物标志物的影响,反映了多组分的特点,多目标,和多途径机制。
Fever is one of the most common clinical conditions and is characterized by pyrogenic infection, malignancy, inflammation, and tissue damage, among others. Ellagic acid (EA) can inhibit the expression of related proteins on the pathway by blocking the nuclear factor kappa-B(NF-κB) signaling pathway, inhibit the levels of pro-inflammatory factors interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α), increase the level of anti-inflammatory factor IL-10, and effectively alleviate inflammatory symptoms. In addition, EA can also reduce the levels of malondialdehyde(MDA) and nitric oxide(NO) in the body, increase the activities of superoxide dismutase (SOD), glutathione (GSH), and catalase(CAT), scavenge oxidative free radicals, inhibit lipid oxidation, and achieve antipyretic and anti-inflammatory effects. The purpose of this study was to establish the relationship between EA and various inflammatory markers, such as TNF-α, IL-6, IL-1β, prostaglandin E2(PGE2), and cyclic adenosine monophosphate(cAMP), and clarify the mechanism of the cyclooxidase-2(COX-2)/NF-κB signaling pathway. Combined with the metabolomics analysis, our study revealed the effects of EA on multiple endogenous biomarkers, reflecting the characteristics of a multi-component, multi-target, and multi-pathway mechanism. Compared to lipopolysaccharide (LPS)- treated animals, subsequent administration of EA significantly lowered the LPS-induced rectal temperature increase (p < 0.05 or p < 0.01), significantly increased serum SOD and GSH levels (p < 0.05 or p < 0.01), and significantly decreased serum MDA, IL-1β, IL-6, and TNF-α levels (p < 0.05 or p < 0.01). In addition, compared to LPS-treated animals, subsequent administration of EA significantly decreased cerebrospinal fluid cAMP and PGE2 levels (p < 0.05 or p < 0.01), significantly decreased cAMP, significantly increased 5-HT levels (p < 0.05 or p < 0.01), and significantly down-regulated p-NF-κB p65 and COX-2 protein levels in the hypothalamus. Subsequent gas chromatography mass spectrometry(GC-MS) metabolite analysis indicated that 12 differential metabolites were detected in serum isolated 4 h after LPS treatment, and 10 differential metabolites were detected in serum collected 7 h after LPS treatment. Next, Pearson correlation analysis was used to systematically characterize the relationship between the identified metabolites and TNF-α, IL-6, MDA, SOD, PGE2, and cAMP. The levels of propionic acid, pyridine, and L-valine were up-regulated by EA, which inhibited the expression of MDA, IL-1β, and TNF-α and increased the activity of GSH. The levels of inositol, urea, and 2-monopalmitin were down-regulated by EA, which inhibited the expression of MDA, IL-1β, and TNF-α, increased the activity of SOD and GSH, reduced the inflammatory response, and alleviated the oxidative stress state. Combined with the results of the metabolic pathway analysis, we suggest that the pathways of the galactose metabolism, synthesis and degradation of ketone bodies, as well as ascorbic acid and aldehyde acid metabolism are closely related to the antipyretic and anti-inflammatory effects of EA. Our study established the relationship between EA and various inflammatory markers, such as TNF-α, IL-6, IL-1β, PGE2, and cAMP, and clarified the mechanism of the COX-2/NF-κB signaling pathway. Combined with the metabolomics analysis, our study revealed the effects of EA on multiple endogenous biomarkers, reflecting the characteristics of a multi-component, multi-target, and multi-pathway mechanism.