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中国精品科技期刊2020
侯佳丽,朱珍珠,谢旻浩,等. 可溶性青梅果胶的表征及其对脂多糖诱导的RAW246.7巨噬细胞炎症的抑制作用[J]. 食品工业科技,2022,43(19):401−409. doi: 10.13386/j.issn1002-0306.2022010013.
引用本文: 侯佳丽,朱珍珠,谢旻浩,等. 可溶性青梅果胶的表征及其对脂多糖诱导的RAW246.7巨噬细胞炎症的抑制作用[J]. 食品工业科技,2022,43(19):401−409. doi: 10.13386/j.issn1002-0306.2022010013.
HOU Jiali, ZHU Zhenzhu, XIE Minhao, et al. Characteristics of Water-soluble Pectin from Greengage and Its Anti-inflammatory Activity in LPS-induced RAW246.7 Cells[J]. Science and Technology of Food Industry, 2022, 43(19): 401−409. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022010013.
Citation: HOU Jiali, ZHU Zhenzhu, XIE Minhao, et al. Characteristics of Water-soluble Pectin from Greengage and Its Anti-inflammatory Activity in LPS-induced RAW246.7 Cells[J]. Science and Technology of Food Industry, 2022, 43(19): 401−409. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022010013.

可溶性青梅果胶的表征及其对脂多糖诱导的RAW246.7巨噬细胞炎症的抑制作用

Characteristics of Water-soluble Pectin from Greengage and Its Anti-inflammatory Activity in LPS-induced RAW246.7 Cells

  • 摘要: 本研究从青梅果汁中提取可溶性果胶(WSP),通过高效液相色谱(HPLC)、高效凝胶色谱(HPGFC)、傅里叶变换红外光谱(FT-IR)、扫描电镜(SEM)及X-射线衍射(XRD)等对其组成和结构进行了表征,并用酶联免疫法(ELISA)对脂多糖(LPS)诱导的RAW246.7细胞的抗炎活性进行评价。结果表明,WSP的酯化度为18.89%,属于低酯果胶,其半乳糖醛酸(GalA)含量为69.72%,主要中性多糖是半乳糖(Gal)(18.36%),重均分子量(Mw)为353.73 kDa,属于非晶型结构。WSP可显著抑制LPS诱导的RAW264.7细胞中肿瘤坏死因子α(TNF-α)(P<0.05)、干扰素-γ(IFN-γ)(P<0.05)、白介素6(IL-6)(P<0.001)的释放量,增加白介素10(IL-10)(P<0.01)的分泌,起到抗炎作用。为了探究其抗炎机理,通过实时荧光定量聚合酶链式反应法(Real-time qPCR)和蛋白印迹法(Western blot)研究了WSP对Janus 激酶/信号转导和转录激活因子信号通路(JAK/STAT信号通路)的影响。与LPS模型组相比,WSP显著上调了与炎症密切相关的JAK1(P<0.01),JAK2(P<0.05),JAK3(P<0.001)和STAT1(P<0.001),STAT2(P<0.001)和STAT3(P<0.05)的mRNA表达量,且使JAK1和STAT3的磷酸化水平显著降低(P<0.05),该结果表明WSP可通过干预JAK/STAT信号通路,抑制与炎症信号相关的JAK和STAT家族的磷酸化水平来缓解LPS诱导的RAW264.7细胞炎症。本研究可为评价青梅果的健康促进作用和提高青梅果加工附加值提供参考。

     

    Abstract: In this study, the water soluble pectin (WSP) from greengage juice was extracted. The structure and composition of WSP were characterized by high performance liquid chromatography (HPLC), high performance gel filtration chromatography (HPGFC), Fourier transform infrared spectrometer (FT-IR), scanning electron microscope (SEM) and X-ray diffraction (XRD). And the anti-inflammatory activity of WSP in lipopolysaccharide (LPS)-induced RAW246.7 cells was evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that WSP was a low-methoxy pectin with DE value of 18.89%. The content of galacturonic acid (GalA) was 69.72%, and the galactose (Gal) was the predominant neutral sugar with the content of 18.36%. The molecular weight (Mw) of WSP was 353.73 kDa. WSP significantly inhibited the release of tumor necrosis factor α (TNF-α) (P<0.05), interferon-γ (IFN-γ) (P<0.05), interleukin-6 (IL-6) (P<0.001) and increasing the secretion of interleukin-10 (IL-10) (P<0.01). The effects of WSP on Janus kinase/signal transduction and transcription activator signaling pathway (JAK/STAT signaling pathway) were investigated by real-time quantitative PCR and Western blot in order to clarify the anti-inflammatory mechanism. Compared with LPS group, WSP significantly upregulated the mRNA expressions of JAK1 (P<0.01), JAK2 (P<0.05), JAK3 (P<0.001), STAT1 (P<0.001), STAT2 (P<0.001) and STAT3 (P<0.05). Further analysis showed that the phosphorylation levels of JAK1 and STAT3 were significantly inhibited by WSP treatment (P<0.05). Therefore, the WSP can alleviate LPS-induced inflammation in RAW264.7 cells by interfering with JAK/STAT signaling pathway and inhibiting the phosphorylation levels of JAK and STAT families related to inflammatory signaling. This study can provide a basis for the evaluation of the health improvement effect of greengage fruit and the development of value added greengage product.

     

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