首页|白术内酯Ⅲ介导的肌动蛋白相关蛋白2/3抑制肠上皮间质转化以缓解溃疡性结肠炎的研究

白术内酯Ⅲ介导的肌动蛋白相关蛋白2/3抑制肠上皮间质转化以缓解溃疡性结肠炎的研究

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原文链接
  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
目的:探寻白术内酯Ⅲ(AT-Ⅲ)的关键靶点,探究肌动蛋白相关蛋白2/3(Arp2/3)抑制溃疡性结肠炎(UC)的作用机制.方法:采用有限酶切-质谱法(lip-SMap)筛选大鼠小肠隐窝上皮细胞(IEC-6)特征蛋白,进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,确定AT-Ⅲ的关键靶点.脂多糖(LPS)与IEC-6共孵育模拟UC模型,酶联免疫吸附实验(ELISA)检测白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α)含量;划痕实验测定IEC-6迁移速率;蛋白质印迹法测定上皮钙黏素(E-cadherin)、波形蛋白(Vimentin)和Arp2/3含量;分子对接评估AT-Ⅲ与关键靶点的结合能力.结果:基于lip-SMap获得的特征蛋白参与调控磷脂酰肌醇3激酶-丝苏氨酸激酶(PI3K-AKT)信号通路和紧密连接(TJ)信号通路;与对照组比较,LPS组中IL-6和TNF-α释放量升高(均P＜0.05),迁移速率变快(P＜0.05),E-cadherin和Vimentin的表达都有不同程度的影响(均P＜0.05);与LPS组比较,LPS+CK666+AT-Ⅲ组显著降低IL-6和TNF-α含量(均P＜0.01),有效逆转LPS诱导的细胞迁移变化及E-cadherin和Vimentin表达(均P＜0.05);分子对接显示,Arp2/3与AT-Ⅲ有较好的结合活性.结论:Arp2/3作为AT-Ⅲ的关键靶点,可通过上皮细胞-间充质转化(EMT)缓解LPS诱导的UC.
Atractylenolide Ⅲ-mediated Actin-related Protein 2/3 Alleviates Ulcerative Colitis by Inhibiting Intestinal Epithelial-mesenchymal Transition
Objective:To identify the key targets of atractylenolide Ⅲ(AT-Ⅲ)and to explore the mechanism of actin-related pro-tein 2/3(Arp2/3)in inhibiting ulcerative colitis(UC).Methods:Characteristic proteins of rat small intestinal crypt epithelial cells(IEC-6)were screened using limited proteolysis-mass spectrometry(lip-SMap).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to determine the key targets of AT-Ⅲ.An UC model was simula-ted by co-incubating lipopolysaccharide(LPS)with IEC-6 cells.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α).A scratch assay was performed to measure the migration rate of IEC-6 cells.Western blot was used to measure the levels of E-cadherin,vimentin,and Arp2/3.Molecular docking was con-ducted to evaluate the binding ability of AT-Ⅲ to its key targets.Results:The characteristic proteins obtained from lip-SMap were involved in regulating the phosphatidylinositol 3-kinase-serine/threonine kinase(PI3K-AKT)signaling pathway and the tight junc-tion(TJ)signaling pathway.Compared with the control group,the LPS group showed increased levels of IL-6 and TNF-α(both P＜0.05),an increased migration rate(P＜0.05),and various impacts on the expression of E-cadherin and vimentin(both P＜0.05).Compared with the LPS group,the LPS+CK666+AT-Ⅲ group significantly reduced IL-6 and TNF-α levels(both P＜0.01),ef-fectively reversed LPS-induced changes in cell migration and the expression of E-cadherin and vimentin(both P＜0.05).Molecular docking showed that Arp2/3 had good binding activity with AT-Ⅲ.Conclusion:Arp2/3 is a key target of AT-Ⅲ and can alleviate LPS-induced UC through epithelial-mesenchymal transition(EMT).

Atractylenolide ⅢIEC-6Ulcerative colitisActin-related protein 2/3Limited proteolysis-mass spectrometryEpi-thelial-mesenchymal transitionTight junction pathwayLipopolysaccharide

肖贵梅、蔡向志、龙海洮、田丽霞、任燕

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贵州大学药学院,贵阳,550000

白术内酯Ⅲ 大鼠小肠隐窝上皮细胞 溃疡性结肠炎 肌动蛋白相关蛋白2/3 有限酶切-质谱法 上皮细胞-间充质转化 紧密连接通路 脂多糖

国家自然科学基金项目贵州省科技计划项目

82360754黔科合LH字[2017]7295号

2024

10.3969/j.issn.1673-7202.2024.15.004

世界中医药
世界中医药学会联合会

世界中医药

CSTPCDCHSSCD北大核心
影响因子：1.266
ISSN：1673-7202
年,卷(期)：2024.19(15)