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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (3): 279-282.doi: 10.12047/j.cjap.5931.2020.061

• 研究论文 • 上一篇    下一篇

神经酰胺合酶-神经酰胺通路在青蒿琥酯抑制肝纤维化中的作用

阮建佳1△, 杜岩2   

  1. 1. 天津市蓟州区人民医院综合内科,
    2. 天津市蓟州区人民医院药剂科, 天津 301900
  • 收稿日期:2019-08-29 修回日期:2020-03-26 发布日期:2020-09-25
  • 通讯作者: Tel: 022-60733012; E-mail: wangyiaruan@163.com

Effects of artesunate on the inhibition of hepatic fibrosis through ceramide synthase-ceramide pathway

RUAN Jian-jia1△, DU Yan2   

  1. 1. Department of Comprehensive Internal Medicine,
    2. Department of Pharmacy, Tianjin Jizhou District People's Hospital, Tianjin 301900, China
  • Received:2019-08-29 Revised:2020-03-26 Published:2020-09-25

摘要: 目的: 探讨神经酰胺通路在青蒿琥酯(Art)抑制肝纤维化过程中的作用。方法: 将肝星状细胞(LX-2)分为细胞对照组、Art 350 μmol/L组、神经酰胺合酶阻断剂(FB1) 6 μmol/L组及FB1 6 μmol/L + Art 350 μmol/L合用组。每组7个复孔,作用24 h后,收集细胞及上清液进行检测。Western blot法测定神经酰胺合酶2(LASS2)、过氧化物酶体增殖物激活受体-γ(PPAR-γ)、Caspase-3蛋白的表达, HPLC-FLD法对神经酰胺(Cer)的含量进行测定,MTT法检测LX-2的增殖情况,酶消化法检测羟脯氨酸(Hyp)的含量。结果: 与细胞对照组比较,Art处理组神经酰胺合酶的蛋白表达、神经酰胺含量显著增加、LX-2细胞的增殖明显抑制、PPAR-γ和Caspase-3蛋白表达上调、羟脯氨酸分泌抑制 (P<0.05);FB1处理组神经酰胺合酶的蛋白表达和神经酰胺含量显著降低、LX-2细胞增殖显著增加、PPAR-γ和Caspase-3蛋白表达下调、羟脯氨酸分泌增加(P<0.05);与Art单用组比较,两药合用可显著降低Art对神经酰胺合酶蛋白的表达和神经酰胺含量升高的作用,减弱Art对LX-2细胞增殖、PPAR-γ、Caspase-3蛋白表达及羟脯氨酸分泌的作用(P<0.05)。结论: 青蒿琥酯可通过神经酰胺合酶-神经酰胺通路增加细胞中神经酰胺水平,发挥抑制肝纤维化的作用。

关键词: 肝纤维化, 神经酰胺, 神经酰胺合酶, 青蒿琥酯, 肝星状细胞

Abstract: Objective: To investigate the effects of ceramide pathway on the inhibition of artesunate (Art) to hepatic fibrosis.Methods: LX-2 cells were divided into control group, Art treated group with 350 μmol/L, fumonisin B1 (FB1) treated group with 6 μmol/L, and Co-administration group of artesunate 350 μmol/L and fumonisin B1 6 μmol/L. There were 7 compound holes in each group. After 24 hours of treatment, the cells and supernatant were collected and detected. The expressions of homo sapiens longevity assurance homologue 2 (LASS2), peroxisome proliferators-activated receptors-γ (PPAR-γ) and Caspase-3 were evaluated by Western blot, the content of ceramide was evaluated by HPLC-FLD method, MTT assay was adopted to measure the rate of proliferation of LX-2 cells. The content of hydroxyproline was determined by digestive method. Results: Compared with the control group, the expression of ceramide synthase protein and the ceramide content were increased significantly, the proliferation of LX-2 cells was inhibited significantly, the expressions of PPAR-γ and Caspase-3 protein were up-regulated and the secretion of hydroxyproline was inhibited in Art treated group (P<0.05). In FB1 treated group, the protein expression of ceramide synthase and the ceramide content were decreased significantly, the proliferation of LX-2 cells was increased significantly, the expressions of PPAR-γ and Caspase-3 protein were down-regulated, and the secretion of hydroxyproline was increased (P<0.05). Compared with the Art alone group, the combination of the two drugs could significantly reduce the effects of Art on the expression of ceramide synthase protein and the increase of ceramide content, and attenuate the effects of Art on the cell proliferation , PPAR-γ, Caspase-3 protein expression and hydroxyproline level of LX-2 cells (P<0.05). Conclusion: Artesunate could inhibit hepatic fibrosis by increasing the content of ceramide through the ceramide synthase-ceramide pathway.

Key words: hepatic fibrosis, ceramide, synthase-ceramide, artesunate, LX-2

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