Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-<italic style="font-style: italic">κ</italic>B Signaling Pathway

ZHANG Hongjie; SU Dan; ZHU Genhua; SONG Yonggui; ZHOU Bugao; LI Shanshan; ZHANG Changhua; AI Zhifu

doi:10.13422/j.cnki.syfjx.20222138

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Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway

更新时间：2022-10-18

- Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 22, Pages: 58-67(2022)
- 作者机构：
  
  江西中医药大学中医脑科学研究所，江西省中医药防治认知障碍脑疾病重点实验室，江西省中医药管理局抑郁症中医证候动物模型重点研究室，江西省中医药管理局中药药效（防治精神障碍脑疾病）评价重点研究室，南昌 330006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20222138
  CLC： R2-0;R33;R289;R745.1
- Received：08 May 2022，
  
  Published Online：30 August 2022，
  
  Published：20 November 2022
- 稿件说明：
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张红杰,苏丹,朱根华等.黄芩-黄连抗神经炎症的配伍优势及其调控TLR4/MyD88/NF-κB信号通路的潜在靶点[J].中国实验方剂学杂志,2022,28(22):58-67.

ZHANG Hongjie,SU Dan,ZHU Genhua,et al.Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(22):58-67.
张红杰,苏丹,朱根华等.黄芩-黄连抗神经炎症的配伍优势及其调控TLR4/MyD88/NF-κB信号通路的潜在靶点[J].中国实验方剂学杂志,2022,28(22):58-67. DOI： 10.13422/j.cnki.syfjx.20222138.

ZHANG Hongjie,SU Dan,ZHU Genhua,et al.Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(22):58-67. DOI： 10.13422/j.cnki.syfjx.20222138.

摘要

目的

探索黄芩-黄连防治神经炎症的配伍优势，并基于Toll样受体4（TLR4）/髓样分化因子88（MyD88）/核转录因子-

B（NF-

B）通路，阐明黄芩-黄连配伍优势的作用特点及机制。

方法

选取体外具有代表性的小鼠小胶质细胞（BV2），将BV2分为8组，分别为正常组、模型组、黄芩-黄连组、吡拉西坦组、黄芩组、黄连组；利用细菌脂多糖（LPS）建立BV2细胞炎症模型，通过细胞增殖活性检测（CCK-8）试剂盒检测细胞活性；明场下观察细胞形态；酶联免疫吸附测定法（ELISA）、免疫荧光法测定药物对BV2细胞促炎因子产生及释放的影响；实时荧光定量聚合酶链式反应（Real-time PCR）检测TLR4、MyD88、NF-

B mRNA表达；细胞免疫荧光检测NF-

B p65核转位情况；通过TLR4信号转导抑制剂CLI-095、NF-

B阻断剂PDTC进行黄芩-黄连抗神经炎症作用靶点的确认。

结果

与正常组比较，LPS诱导的模型组细胞大多处于激活状态，培养液及细胞内的IL-6、TNF-

、IL-1

含量显著升高（

0.01），TLR4、MyD88、NF-

B p50及NF-

B p65的mRNA表达显著升高（

0.01），NF-

B p65的入核现象明显；与模型组比较，黄芩-黄连、吡拉西坦组预处理后，BV2细胞形态大部分恢复，IL-6、TNF-

、IL-1

水平均显著降低（

0.01），TLR4、MyD88、NF-

B p50及NF-

B p65 mRNA表达明显降低（

0.05，

0.01），NF-

B p65大多存在于细胞质中；黄芩组、黄连组与模型组比较，细胞形态略有恢复，促炎因子水平及TLR4、MyD88、NF-

B p50、NF-

B p65 mRNA表达量有所降低；与黄芩-黄连组比较，黄芩组、黄连组的促炎因子抑制作用效果显著低于药对组；与模型组比较，应用信号通路特异抑制剂“黄芩-黄连+CLI-095”组、“黄芩-黄连+PDTC”组，能够明显降低TLR4、MyD88、NF-

B p50及NF-

B p65的mRNA表达（

0.05，

0.01），并明显抑制NF-

B p65转入细胞核内。

结论

黄芩与黄连相须配伍后的抗神经炎症作用明显优于单味黄芩或黄连；该药对的抗神经炎症优势与抑制小胶质细胞中的TLR4/MyD88/NF-

B信号通路激活密切相关；通过验证说明TLR4、MyD88、NF-

B为黄芩-黄连发挥药对配伍优势的潜在靶点。

Abstract

Objective

To explore the compatibility advantage of Scutellariae Radix-Coptidis Rhizoma in the prevention and treatment of neuroinflammation， and to elucidate the action characteristics and mechanism of the compatibility advantage based on Toll like receptor （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor kappaB （NF-

B） pathway.

Method

Representative mouse microglia cells （BV2）

in vitro

were selected and divided into 8 groups： control group， model group， Scutellariae Radix-Coptidis Rhizoma group， Piracetam group， Scutellariae Radix group and Coptidis Rhizoma group. The BV2 cell inflammatory model was established by lipopolysaccharide （LPS）， and the cell activity was detected by cell counting kit-8 （CCK-8）. Cell morphology was observed under bright field. The production and release of pro-inflammatory factors in BV2 cells were determined by enzyme-linked immunosorbent assay （ELISA） and immunofluorescence assay， and the mRNA expressions of TLR4， MyD88 and NF-

B were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The nuclear translocation of NF-

B p65 was detected by immunofluorescence， and TLR4 signal transduction inhibitor （CLI-095） and NF-

B inhibitor （PDTC） were used to confirm the anti-neuroinflammation targets of Scutellariae Radix-Coptidis Rhizoma.

Result

Compared with the conditions in the control group， most cells in LPS-induced model group were activated， and the contents of IL-6， TNF-

and IL-1

in culture medium and cells and the mRNA expressions of TLR4， MyD88， NF-

B p50 and NF-

B p65 were increased （

0.01）， with obvious nuclear entry of NF-

B p65. Compared with the conditions in the model group， BV2 cell morphology was mostly recovered after pretreatment in Scutellariae Radix-Coptidis Rhizoma and Piracetam groups， and the levels of IL-6， TNF-

and IL-1

and the mRNA expressions of TLR4， MyD88， NF-

B p50 and NF-

B p65 were decreased （

0.05，

0.01）， with NF-

B p65 mostly observed in cytoplasm. Compared with the conditions in the model group， cell morphology was slightly recovered in Scutellariae Radix group and Coptidis Rhizoma group， and the levels of pro-inflammatory factors and mRNA expressions of TLR4， MyD88， NF-

B p50 and NF-

B p65 were reduced. In terms of inhibitory effect on pro-inflammatory factors， Scutellariae Radix group and Coptidis Rhizoma group were lower than Scutellariae Radix-Coptidis Rhizoma group （

0.05）. Compared with the model group， the "Scutellariae Radix-Coptidis Rhizoma+CLI-095" group and "Scutellariae Radix-Coptidis Rhizoma+PDTC" group had lowered mRNA expressions of TLR4， MyD88， NF-

B p50 and NF-

B p65 （

0.05，

0.01）， and the transfer of NF-

B p65 into nucleus was obviously inhibited.

Conclusion

The anti-neuroinflammation effect of Scutellariae Radix-Coptidis Rhizoma was significantly better than Scutellariae Radix or Coptidis Rhizom alone， and the anti-neuroinflammation advantage was closely related to the inhibition of activation of TLR4/MyD88/NF-

B signaling pathway in microglial cells. It was confirmed that TLR4， MyD88 and NF-

B were potential targets for Scutellariae Radix-Coptidis Rhizoma to exert the compatibility advantage.

关键词

Keywords

references

INOUE K . Potential significance of CX3CR1 dynamics in stress resilience against neuronal disorders ［J］. Neural Regen Res ， 2022 ， 17 （ 10 ）： 2153 .

ERDOGAN M A ， KIRAZLAR M ， YIGITTURK G ， et al . Digoxin exhibits neuroprotective properties in a rat model of dementia ［J］. Neurochem Res ， 2022 ， 47 （ 5 ）： 1290 .

ZHANG P F ， GAO F . Neuroinflammation in Parkinson's disease：A meta-analysis of PET imaging studies ［J］. J Neurol ， 2022 ， 269 （ 5 ）： 2304 .

王维 . 转位分子蛋白在脂多糖致小鼠中枢神经炎症中的调控作用及机制研究［D］. 北京：解放军总医院， 2017 .

李涛 . 脂多糖对BV-2小胶质细胞的影响及吡咯烷二硫代氨基甲酸的干预效应［D］. 广州：广州医科大学， 2018 .

邵园，王建，许海云 . 精神分裂症的神经炎症机制［J］. 临床精神医学杂志， 2016 ， 26 （ 4 ）： 3 .

ZHANG G X ， ZHOU A L ， ZHANG G P ， et al . The role of TLR4-mediated MyD88-dependent pathway in neuroinflammation in hippocampal neurons of rats ［J］. Chin J Appl Physiol ， 2013 ， 29 （ 1 ）： 42 .

金毅然，邵宝平，丁艳平，等 . TLR4在神经炎症及其介导的神经退行性疾病中的作用［J］. 中国免疫学杂志， 2019 ， 35 （ 12 ）： 1524 - 1527，1532 .

DUAN H ， QU L ， SHOU C . Mycoplasma hyorhinis induces epithelial-mesenchymal transition in gastric cancer cell MGC803 via TLR4-NF- κ B signaling ［J］. Cancer Lett ， 2014 ， 354 （ 2 ）： 447 .

刘思颖，邝枣园，张韧，等 . 黄芩苷对代谢性炎症及肠道菌群的调节作用探讨［J］. 广州中医药大学学报， 2016 ， 33 （ 3 ）： 372 .

HE X ， WEI Z ， ZHOU E ， et al . Baicalein attenuates inflammatory responses by suppressing TLR4 mediated NF- κ B and MAPK signaling pathways in LPS-induced mastitis in mice ［J］. Int Immunopharmacol ， 2015 ， 28 （ 1 ）： 470 - 476 .

WANG X ， WANG N ， LI H ， et al . Up-regulation of PAI-1 and down-regulation of uPA are involved in suppression of invasiveness and motility of hepatocellular carcinoma cells by a natural compound berberine ［J］. Int J Mol Sci ， 2016 ， 17 （ 4 ）： 577 .

庞婕 . 黄连活性物质的抗焦虑作用及其机理研究［D］. 重庆：西南大学， 2014 .

ASKI M L ， REZVANI M E ， KHAKSARI M ， et al . Neuroprotective effect of berberine chloride on cognitive impairment and hippocampal damage in experimental model of vascular dementia ［J］. Iran J Basic Med Sci ， 2018 ， 21 （ 1 ）： 53 - 58 .

CHEN C C ， HUNG T H ， LEE C Y ， et al . Berberine protects against neuronal damage via suppression of glia-mediated inflammation in traumatic brain injury ［J］. PLoS One ， 2014 ， 9 （ 12 ）： e115694 .

刘志勇 . 加味葛根芩连汤治疗阳明湿热瘀阻型血管性痴呆临床疗效观察［J］. 海峡药学， 2015 ， 27 （ 6 ）： 241 - 242 .

高炎 . 黄连解毒汤治疗阿尔茨海默病的临床观察及实验研究［D］. 武汉：湖北中医药大学， 2021 .

INOUE K . Potential significance of CX3CR1 dynamics in stress resilience against neuronal disorders ［J］. Neural Regen Res ， 2022 ， 17 （ 10 ）： 2153 .

章常华，刘彤彤，邓可众，等 . 黄芩-黄连防治 D -半乳糖痴呆小鼠的作用机制［J］. 中成药， 2018 ， 40 （ 3 ）： 524 - 529 .

DANIELA G . Inflammation and oxidative damage in Alzheimer's disease：Friend or foe？［J］. Frontiers in Bioscience ， 2011 ， 3 （ 1 ）： 252 .

DYNE E ， CAWOOD M ， SUZELIS M ， et al . Ultrastructural analysis of the morphological phenotypes of microglia associated with neuroinflammatory cues ［J］. J Comp Neurol ， 2022 ， 530 （ 8 ）： 1263 .

TAKI-NAKANO N ， KOTERA J ， OHTA H . 12-oxo-phytodienoic acid，a plant-derived oxylipin，attenuates lipopolysaccharide-induced inflammation in microglia ［J］. Biochem Biophys Res Commun ， 2016 ， 473 （ 4 ）： 1288 .

张丽丽，段淑荣，赵敬堃，等 . Toll样受体4在乙醇所致的小胶质细胞激活中的作用［J］. 中国神经免疫学和神经病学杂志， 2011 ， doi： 10.3969/j.issn.1006-2963.2011.06.017 http://dx.doi.org/10.3969/j.issn.1006-2963.2011.06.017 .

YUAN L ， LIU S ， BAI X ， et al . Oxytocin inhibits lipopolysaccharide-induced inflammation in microglial cells and attenuates microglial activation in lipopolysaccharide-treated mice ［J］. J Neuroinflammation ， 2016 ， 13 （ 1 ）： 77 .

BAKKAR N ， GUTTRIDGE D C . NF-kappaB signaling：A tale of two pathways in skeletal myogenesis ［J］. Physiol Rev ， 2010 ， 90 （ 2 ）： 495 - 511 .

LU Y C ， YEH W C ， OHASHI P S . LPS/TLR4 signal transduction pathway ［J］. Cytokine ， 2008 ， 42 （ 2 ）： 145 - 151 .

吕玉华 . CagA与YWHAE互作激活NF- κ B的途径及细胞侵袭转移效应［D］. 福州：福建医科大学， 2018 .

FAYEZ A M ， ELNOBY A S ， BAHNASAWY N H ， et al . Neuroprotective effects of zafirlukast，piracetam and their combination on L -methionine-induced vascular dementia in rats ［J］. Fundam Clin Pharmacol ， 2019 ， 33 （ 6 ）： 634 - 648 .

ÇAKICI N ， JUDGE-HUNDAL G ， KOOLA M M ， et al . An update on the efficacy of anti-inflammatory agents for patients with schizophrenia：A Meta-analysis ［J］. Psychol Med ， 2019 ， 49 （ 14 ）： 2307 - 2319 .

ZAITONE S A ， ABO-ELMATTY D M ， ELSHAZLY S M . Piracetam and vinpocetine ameliorate rotenone-induced Parkinsonism in rats ［J］. Indian J Pharmacol ， 2012 ， 44 （ 6 ）： 774 - 779 .

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Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway

DOI：10.13422/j.cnki.syfjx.20222138

摘要

Abstract

关键词

Keywords

references