Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells

LIU Yaqing; LIU Bin; MA Xuran; WANG Dunfang; FENG Xue; YANG Weipeng

doi:10.13422/j.cnki.syfjx.20221704

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Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells

更新时间：2023-05-17

- Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 7, Pages: 20-28(2023)
- 作者机构：
  
  1.中国中医科学院中药研究所，北京 100700
  2.黑龙江中医药大学，哈尔滨 150040
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20221704
  CLC： R2-0;R22;R285.5;R289;R33
- Published：05 April 2023，
  
  Published Online：31 August 2022，
  
  Received：20 June 2022，
- 稿件说明：
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刘雅清,刘滨,马旭冉等.黄芩汤对LPS诱导的RAW264.7炎症细胞的抗炎作用[J].中国实验方剂学杂志,2023,29(07):20-28.

LIU Yaqing,LIU Bin,MA Xuran,et al.Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(07):20-28.
刘雅清,刘滨,马旭冉等.黄芩汤对LPS诱导的RAW264.7炎症细胞的抗炎作用[J].中国实验方剂学杂志,2023,29(07):20-28. DOI： 10.13422/j.cnki.syfjx.20221704.

LIU Yaqing,LIU Bin,MA Xuran,et al.Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(07):20-28. DOI： 10.13422/j.cnki.syfjx.20221704.

摘要

目的

以RAW264.7细胞为炎症模型，探究黄芩汤的抗炎作用机制。

方法

制备黄芩汤，筛选对RAW264.7细胞的安全剂量；将RAW264.7细胞接种于24孔板中，先后加入黄芩汤和脂多糖（LPS），分别采用Griess法和酶联免疫吸附测定法（ELISA）测定一氧化氮（NO）和白细胞介素-6（IL-6）、肿瘤坏死因子-

（TNF-

）、前列腺素E

（PGE

）的含量；将RAW264.7细胞接种于6孔板中，设空白组、LPS组、LPS+黄芩汤组、核转录因子-

B p65（NF-

B p65）抑制剂（PDTC）组、p38丝裂原活化蛋白激酶（p38 MAPK）抑制剂（SB203580）组、细胞外信号调节激酶（ERK）抑制剂（PD98059）组、c-Jun氨基末端激酶（JNK）抑制剂（SP600125）组、Janus酪氨酸蛋白激酶（JAK）抑制剂（AG490）组，先后加入相应的抑制剂和黄芩汤，并经LPS刺激后，提取RNA和蛋白，分别采用实时荧光定量聚合酶链式反应（Real-time PCR）和蛋白免疫印迹法（Western blot）检测NF-

B p65、p38 MAPK、ERK、JNK和JAK mRNA及蛋白的表达水平，探究黄芩汤通过调控NF-

B、丝裂原活化蛋白激酶（MAPK）和JAK/信号转导及转录激活因子（STAT）信号通路发挥抗炎作用的机制。

结果

与空白组比较，LPS刺激后，模型组细胞中NO、IL-6、TNF-

、PGE

的浓度明显增加（

0.05，

0.01），与模型组比较，加入黄芩汤后，NO、IL-6、TNF-

、PGE

分泌量有减少趋势（

0.05，

0.01）；与空白组比较，模型组细胞内p38 MAPK、ERK、JNK、JAK和NF-

B p65 mRNA及总蛋白表达明显升高（

0.05，

0.01），与模型组比较，黄芩汤孵育后，炎症细胞内p38 MAPK、ERK、JNK、JAK和NF-

B p65总蛋白及mRNA的表达明显降低（

0.05，

0.01）；同时各抑制剂组细胞内NF-

B p65总蛋白及mRNA表达均有下降趋势（

0.05，

0.01）。

结论

黄芩汤能够通过NF-

B、MAPK和JAK/STAT信号通路抑制炎症反应。

Abstract

Objective

To explore the anti-inflammatory mechanism of Huangqintang based on the inflammation model in RAW264.7 cells.

Method

Huangqintang was prepared and the safe dose to RAW264.7 cells was screened out. The RAW264.7 cells were seeded in 24-well plates and incubated with Huangqintang and lipopolysaccharide （LPS）， successively. The concentrations of nitric oxide （NO）， interleukin （IL）-6， tumor necrosis factor （TNF）-

， and prostaglandin E

（PGE

） were measured by Griess assay and enzyme-linked immunosorbent assay （ELISA）， respectively. Meanwhile， RAW264.7 cells were inoculated in 6-well plates， and normal group， LPS group， LPS+Huangqintang group， nuclear factor-

B （NF-

B） p65 inhibitor PDTC group， p38 mitogen-activated protein kinase （MAPK） inhibitor SB203580 group， extracellular signal-regulated kinase （ERK） inhibitor PD98059 group， c-Jun N-terminal kinase （JNK） inhibitor SP600125 group， and Janus kinase （JAK） inhibitor AG490 group were set up. After the cells were incubated with corresponding inhibitors and Huangqintang and stimulated by LPS， RNA and protein were extracted. The mRNA and protein expression levels of NF-

B p65， p38 MAPK， ERK， JNK， and JAK were detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively， to explore the anti-inflammatory mechanism of Huangqintang by regulating the NF-

B， MAPK， and JAK/signal transducer and activator of transcription protein （STAT） signaling pathways.

Result

After stimulation with LPS， the concentrations of NO， IL-6， TNF-

， and PGE

in the cells of the model group increased significantly（

0.05，

0.01）. Compare with the model group， after incubation with Huangqintang， the secretion of NO， IL-6， TNF-

， and PGE

showed a downward trend （

0.05，

0.01）. Compared with the normal group， the model group showed increased mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-

B p65 and total protein expression in cells after stimulation with LPS （

0.05，

0.01）. Compare with the model group，after incubation with Huangqintang， the total protein and mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-

B p65 in inflammatory cells decreased （

0.05，

0.01）. Meanwhile， the expression of NF-

B p65 total protein and mRNA in each inhibitor group showed a downward trend （

0.05，

0.01）.

Conclusion

Huangqintang can inhibit the inflammatory response through the NF-

B， MAPK， and JAK-STAT signaling pathways.

关键词

黄芩汤炎症核转录因子-κB（NF-κB）信号通路丝裂原活化蛋白激酶（MAPK）信号通路Janus酪氨酸蛋白激酶（JAK）/信号转导及转录激活因子（STAT）信号通路

Keywords

Huangqintanginflammationnuclear factor-κB （NF-κB） signaling pathwaymitogen-activated protein kinase （MAPK） signaling pathwayJanus kinase （JAK）/signal transducer and activator of transcription protein （STAT） signaling pathway

references

成梦群，尹健彬，张旋.巨噬细胞自噬在炎症性疾病中的作用研究进展［J］.中国医药导报，2019，16（21）：35-38.

任彩瑗，张向颖，时红波，等.自噬调控脂多糖诱导的巨噬细胞炎症因子水平［J］.细胞与分子免疫学杂志，2017，33（5）：581-585.

汤鑫淼，崔悦，朱鹤云，等.黄芩汤的化学成分与药理作用研究进展［J］.吉林医药学院学报，2022，43（1）：59-61.

王怡薇，张会会，王彦礼，等.黄芩汤对溃疡性结肠炎大鼠NF-κB p65调控作用研究［J］.药学学报，2015，50（1）：21-27.

苗金雪.通过PINK1/Parkin线粒体自噬通路探究黄芩汤对溃疡性结肠炎大鼠的治疗作用［D］.哈尔滨：黑龙江中医药大学，2021.

MA X，WANG D，FENG X，et al.Huangqin tang interference with colitis associated colorectal cancer through regulation of epithelial mesenchymal transition and cell cycle［J］.Front Pharmacol，2022，13：837217.

王德芳，赵明，李晓艳.针刺结合黄芩汤化裁方治疗溃疡性结肠炎活动期（湿热蕴结证）的疗效观察［J］.中医药导报，2022，28（2）：71-75.

杨婧怡，杜康，杨莉，等.不同基原芍药制备的黄芩汤治疗溃疡性结肠炎药效对比研究［J］.中国中药杂志，2021，46（24）：6395-6402.

王敦方.基于TLR4/MyD88通路和组学研究黄芩汤治疗溃疡性结肠炎的作用机制［D］.北京：中国中医科学院，2017.

宋红新.基于网络药理学的黄芩汤调控UC模型小鼠Th17/Treg、Th1/Th2细胞平衡的免疫学机制研究［D］.北京：中国中医科学院，2021.

李涛，王怡薇，王彦礼，等.黄芩汤血浆中多成分LC-MS法测定及其药代动力学特征研究［J］.药学学报，2013，48（6）：917-924.

MEDZHITOV R.Origin and physiological roles of inflammation［J］.Nature，2008，454（7203）：428-435.

MONJE M L，TODA H，PALMER T D.Inflammatory blockade restores adult hippocampal neurogenesis［J］.Science，2003，302（5651）：1760-1765.

ZHANG W J，WEI H，HAGEN T，et al.Alpha-lipoic acid attenuates LPS-induced inflammatory responses by activating the phosphoinositide 3-kinase/Akt signaling pathway［J］.Proc Natl Acad Sci U S A，2007，104（10）：4077-4082.

DI LORENZO F，DE CASTRO C，SILIPO A，et al.Lipopolysaccharide structures of Gram-negative populations in the gut microbiota and effects on host interactions［J］.FEMS Microbiol Rev，2019，43（3）：257-272.

ARANGO DUQUE G，DESCOTEAUX A.Macrophage cytokines： Involvement in immunity and infectious diseases［J］.Front Immunol，2014，5：491.

BANSKOTA S，WANG H，KWON Y H，et al.Salicylates ameliorate intestinal inflammation by activating macrophage AMPK［J］.Inflamm Bowel Dis，2021，27（6）：914-926.

王乐旬，吴惠娟，张盛昔，等.伊马替尼抑制脂多糖诱导的巨噬细胞炎症表型［J］.中山大学学报：医学科学版，2019，40（5）：689-697.

YAN G，CHEN L，WANG H，et al.Baicalin inhibits LPS-induced inflammation in RAW264.7 cells through miR-181b/HMGB1/TRL4/NF-κB pathway［J］.Am J Transl Res，2021，13（9）：10127-10141.

LEE S M，SON K N， SHAH D，et al.Histatin-1 attenuates LPS-induced inflammatory signaling in RAW264.7 macrophages［J］.Int J Mol Sci，2021，22（15）：7856.

HINZ M，SCHEIDEREIT C.The IκB kinase complex in NF-κB regulation and beyond［J］.EMBO Rep，2014，15（1）：46-61.

OECKINGHAUS A，HAYDEN M S，GHOSH S.Crosstalk in NF-κB signaling pathways［J］.Nat Immunol，2011，12（8）：695-708.

ZHANG W，LIU H T.MAPK signal pathways in the regulation of cell proliferation in mammalian cells［J］.Cell Res，2002，12（1）：9-18.

DENT P，YACOUB A，FISHER P B，et al.MAPK pathways in radiation responses［J］.Oncogene，2003，22（37）：5885-5896.

XIN P，XU X，DENG Cet al.The role of JAK/STAT signaling pathway and its inhibitors in diseases［J］.Int Immunopharmacol，2020，80：106210.

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