Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells <italic style="font-style: italic">in vitro</italic>

Jiahang LI; Shengqian YANG; Juanjuan LIU; Mingming LUO; Jie CHEN; Xiaohui LI

doi:10.16016/j.1000-5404.201910149

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Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells in vitro

Basic Medicine | 更新时间：2026-01-09

- Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells in vitro
- Journal of Third Military Medical University Vol. 42, Issue 4, Pages: 342-349(2020)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系药物研究所
  2.264005 山东烟台，烟台大学分子药理和药物评价教育部重点实验室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81473210);the Open Funding Project of Key Laboratory of Molecular Pharmacology and Drug Evaluation of Ministry of Education(P201902)
- DOI：10.16016/j.1000-5404.201910149
  CLC： R282.71; R285.5; R575.2
- Received：22 October 2019，
  
  Revised：2019-12-09，
  
  Published：28 February 2020
- 稿件说明：
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Jiahang LI, Shengqian YANG, Juanjuan LIU, et al. Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(4): 342-349.
DOI：

Jiahang LI, Shengqian YANG, Juanjuan LIU, et al. Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(4): 342-349. DOI： 10.16016/j.1000-5404.201910149.

摘要

目的

探讨连翘苷对脂多糖诱导肝脏星状细胞活化的抑制作用及其机制。

方法

体外培养大鼠肝星状细胞HSC-T6，实验分4组:空白对照组、脂多糖(1 μg/mL)处理组、1 μmol/L连翘苷加脂多糖处理组和10 μmol/L连翘苷加脂多糖处理组。采用MTT法检测HSC-T6细胞的增殖；Transwell实验分析细胞的迁移；流式细胞仪检测细胞内的ROS水平；ELISA法检测细胞上清中促炎细胞因子TNF-α、IL-6、IL-1β的浓度；Western blot分析细胞内α-SMA、NF-κB p65蛋白表达或蛋白磷酸化水平。

结果

浓度为1~100 μmol/L连翘苷对HSC-T6细胞的增殖无明显影响。脂多糖可明显促进HSC-T6细胞增殖及迁移，1 μmol/L和10 μmol/L连翘苷均可显著抑制脂多糖诱导的细胞增殖及迁移，分别降低16.13%、17.14%，48.99%、58.86%。脂多糖处理HSC-T6细胞能明显增加细胞内α-SMA蛋白的表达，而10 μmol/L连翘苷处理后，细胞内α-SMA的表达量减少27.33%。1 μmol/L和10 μmol/L连翘苷对脂多糖诱导细胞内ROS含量(30.03%、51.26%)、促炎细胞因子TNF-α、IL-6、IL-1β(25.16%、39.51%，36.57%、43.75%，58.99%、73.01%)的分泌和NF-κB p65蛋白的磷酸化水平(13.90%、24.96%)也具有显著抑制作用。相关性分析表明:连翘苷处理肝星状细胞后，其降低NF-κB p65磷酸化蛋白表达和对ROS、炎症因子(IL-1β、TNF-α)的作用与其抑制肝星状细胞活化标志物α-SMA蛋白表达的作用呈正相关。

结论

天然产物连翘苷能够显著抑制脂多糖诱导肝星状细胞的活化，其机制可能与其下调NF-κB信号途径介导的炎症反应有关。

Abstract

Objective

To investigate the inhibitory effect of phillyrin (PHI) on lipopolysaccharide (LPS)-induced hepatic stellate cell (HSC) activation and explore the possible mechanisms.

Methods

Cultured rat hepatic stellate cells (HSC-T6) were divided into blank control group

LPS (1 μg/mL) group

LPS (1 μg/mL) + PHI (1 μg/mL) group

and LPS (1 μg/mL) + PHI (10 μg/mL) group. We assessed the changes in the proliferation and migration of the cells (HSC-T6) after the treatments using MTT assay and Transwell migration assay

respectively. The intracellular reactive oxygen species (ROS) level in the cells was determined with flow cytometry

and the concentrations of proinflammatory cytokines in the supernatant were tested using enzyme-linked immunosorbant assay (ELISA). The expression levels of α-SMA and phosphorylated nuclear factor-κB (NF-κB) p65 in the cells were analyzed with Western blotting.

Results

PHI from the concentration of 1 to 100 μmol/L did not significantly affect the proliferation of HSC-T6 cells. LPS stimaulation significantly promoted the proliferation and migration of the cells

and such effects were obviously suppressed by PHI treatment at both 1 (16.13% and 17.14%) and 10 μmol/L(48.99% and 58.86%). PHI at 10 μmol/L significantly reduced the expression of α-SMA (27.33%) in HSC-T6 cells exposed to LPS; at both 1 and 10 μmol/L

PHI strongly suppressed LPS-induced elevation of intracellular ROS level(30.03% and 51.26%)

lowered the levels of proinflammatory cytokines TNF-α(23.16% and 39.51%)

IL-6(36.57% and 43.75%) and IL-1β(58.99% and 73.01%) and decreased the expression of phosphorylated NF-κB p65(13.90% and 24.96%) in LPS-stimulated cells. The decreased expression level of phosphorylated NF-κB p65 protein in HSC-T6 cells was positively correlated with intracellular ROS level

concentrations of TNF-α and IL-1β

and the expression level of HSC activation marker α-SMA protein.

Conclusion

PHI can inhibit HSC activation induced by LPS possibly by down-regulating inflammatory responses mediated by NF-κB signaling pathway.

关键词

Keywords

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Inhibitory effect of phillyrin on lipopolysaccharide-induced activation of rat hepatic stellate cells in vitro

DOI：10.16016/j.1000-5404.201910149

摘要

Abstract

关键词

Keywords

references