| 引用本文: | 周至品,陈勇,吴瑞胜,王竟静,覃乐,黄桂东,刘代华.基于TLR4/NF-κB/NLRP3信号通路研究三叶香茶菜含药血清对枯否细胞活化的影响[J].中国现代应用药学,2023,40(21):2917-2925. |
| ZHOU Zhipin,CHEN Yong,WU Ruisheng,WANG Jingjing,QIN Le,HUANG Guidong,LIU Daihua.Research on the Effect of Isodon Ternifolia-containing Serum on the Activation of Kupffer Cells Based on TLR4/NF-κB/NLRP3 Signal Pathway[J].Chin J Mod Appl Pharm(中国现代应用药学),2023,40(21):2917-2925. |
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| 基于TLR4/NF-κB/NLRP3信号通路研究三叶香茶菜含药血清对枯否细胞活化的影响 |
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周至品1, 陈勇2, 吴瑞胜2, 王竟静2, 覃乐2, 黄桂东2, 刘代华1
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1.广西医科大学附属柳州市人民医院, 广西壮族自治区卫生健康委员会广西临床疾病生物技术研究重点实验室(柳州市人民医院), 柳州市胃肠道中成药工程技术研究中心, 广西 柳州 545006;2.广西中医药大学, 南宁 530200
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| 摘要: |
| 目的 研究三叶香茶菜含药血清(isodon ternifolius-containing serum，ITS)通过Toll样受体4(TLR4)/核因子κB(NF-κB)/NOD样受体蛋白3(NLRP3)信号通路对脂多糖(lipopolysaccharide，LPS)诱导的大鼠原代肝枯否细胞(Kupffer cell，KC)活化的影响。方法 分离培养大鼠原代KC，将LPS诱导的大鼠原代KC分为空白对照组、模型对照组、空白血清组、阳性对照组(秋水仙碱含药血清组)、ITS组、TLR4阻断剂组、TLR4阻断剂+ITS组。MTT法检测不同浓度ITS对KC增殖活性的影响；ELISA法检测KC细胞上清液白介素-1β(IL-1β)、白细胞介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量；荧光定量聚合应(PCR)、Western blotting和免疫荧光检测KC中TLR4/NF-κB/NLRP3信号通路中TLR4、核因子κB抑制蛋白α(IκBα)、半胱肽氨酸蛋白酶-1(Caspase-1)、NLRP3 mRNA和TLR4、IκBα、磷酸化IκBα(p-IκBα)、Caspase-1、NLRP3、NF-κBp65蛋白表达情况。结果 与模型对照组相比，各药物组KC上清液中IL-1β、IL-18、TNF-α和IL-6含量，以及细胞中TLR4、IκBα、Caspase-1、NLRP3 mRNA和TLR4、IκBα、p-IκBα、Caspase-1、NLRP3、NF-κBp65蛋白的表达均下调或降低(P<0.05或P<0.01)；与TLR4阻断剂组比较，TLR4阻断剂+ITS组上述多数指标的改善更加明显。结论 三叶香茶菜可能通过下调TLR4/NF-κB/NLRP3信号通路抑制KC活化，减少炎性因子的表达和释放，从而减轻肝脏炎症损伤。 |
| 关键词: TLR4/NF-κB/NLRP3信号通路 枯否细胞 三叶香茶菜 含药血清 |
| DOI：10.13748/j.cnki.issn1007-7693.20222838 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金项目(81760751)；广西自然科学基金项目(2021GXNSFAA075020)；柳州市科技计划项目(2020NBAB0815)；柳州市人民医院引进高层次人才科研启动基金项目(LRYGCC202104) |
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| Research on the Effect of Isodon Ternifolia-containing Serum on the Activation of Kupffer Cells Based on TLR4/NF-κB/NLRP3 Signal Pathway |
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ZHOU Zhipin1, CHEN Yong2, WU Ruisheng2, WANG Jingjing2, QIN Le2, HUANG Guidong2, LIU Daihua1
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1.Liuzhou People's Hospital Affiliated to Guangxi Medical University, Guangxi Key Laboratory of Clinical Diseases Biotechnology Research of Guangxi Zhuang Autonomous Region Health Commission(Liuzhou People's Hospital), Liuzhou Gastrointestinal Chinese Patent Medicine Engineering and Technology Research Center, Liuzhou 545006, China;2.Guangxi University of Traditional Chinese Medicine, Nanning 530200, China
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| Abstract: |
| OBJECTIVE To investigate the effect of isodon ternifolia-containing serum(ITS) on the activation of rat primary hepatic Kupffer cell(KC) induced by lipopolysaccharide(LPS) through TLR4/NF-κB/NLRP3 signal pathway. METHODS The primary KC of rats were isolated and cultured, and the primary KC induced by LPS were divided into blank control group, model control group, blank serum group, positive control group(colchicine containing serum group), ITS group, TLR4 blocker group and TLR4 blocker+ITS group. MTT assay was used to detect the effect of different concentrations of ITS on the proliferation activity of KC. The content of interleukin-1β(IL-1β), interleukin-18(IL-18), tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in KC supernatant were detected by ELISA. Fluorescence quantitative polymerase chain reaction(PCR), Western blotting and immunofluorescence were used to detect the expression of TLR4, nuclear factor κB inhibitor protein α(IκBα), cysteine protease-1(Caspase-1), NLRP3 mRNA and TLR4, IκBα, phosphorylated IκBα(p-IκBα), Caspase-1, NLRP3 and NF-κBp65 in KC. RESULTS Compared with the model control group, the contents of IL-1β, IL-18, TNF-α and IL-6 in the supernatant of KC and the expression of TLR4, IκB α, Caspase-1, NLRP3 mRNA and TLR4, IκBα, p-IκBα, Caspase-1, NLRP3, NF-κBp65 protein in the supernatant of KC in all drug groups were down-regulated or decreased(P<0.05 or P<0.01). Compared with TLR4 blocker group, the improvement of most of the above indexes in TLR4 blocker+ITS group was more obvious. CONCLUSION Isodon ternifolia may inhibit the activation of KC and reduce the expression and release of inflammatory factors by down-regulating TLR4/NF-κB/NLRP3 signal pathway, thus alleviating the inflammatory injury of liver. |
| Key words: TLR4/NF-κB/NLRP3 signaling pathway Kupffer cell isodon ternifolia drug-containing serum |
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