Effect of Danggui Shaoyaosan Ameliorates in Alleviating H<sub>2</sub>O<sub>2</sub>-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-<italic style="font-style: italic">κ</italic>B Pathway

Jing-ping YU; Chun-xiang HE; Shao-wu CHENG; Yi-xuan CHEN; Zhen-yan SONG

doi:10.13422/j.cnki.syfjx.20200604

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Effect of Danggui Shaoyaosan Ameliorates in Alleviating H₂O₂-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway

Classic Prescriptions | 更新时间：2021-02-09

- Effect of Danggui Shaoyaosan Ameliorates in Alleviating H₂O₂-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 10, Pages: 1-7(2020)
- 作者机构：
  
  湖南中医药大学　中西医结合心脑疾病防治湖南省重点实验室，长沙　 410208
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200604
  CLC： R2-0; R22; R285.5; R289
- Received：04 September 2019，
  
  Published Online：06 December 2019，
  
  Published：20 May 2020
- 稿件说明：
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Jing-ping YU, Chun-xiang HE, Shao-wu CHENG, et al. Effect of Danggui Shaoyaosan Ameliorates in Alleviating H₂O₂-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 1-7.
DOI：

Jing-ping YU, Chun-xiang HE, Shao-wu CHENG, et al. Effect of Danggui Shaoyaosan Ameliorates in Alleviating H₂O₂-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 1-7. DOI： 10.13422/j.cnki.syfjx.20200604.

摘要

目的：

研究当归芍药散(Danggui Shaoyaosan，DSS)含药血清对过氧化氢(H

)诱导人神经母细胞瘤细胞(SH-SY5Y)氧化应激和炎症反应的调控作用。

方法：

采用噻唑蓝(MTT)比色法测定SH-SY5Y细胞存活率构建最佳时间和最佳剂量的H

诱导的细胞损伤模型。实验设空白组，模型组，当归芍药散含药血清高、中、低剂量组(2.5%，5%，10%)。采用MTT比色法检测DSS干预后细胞活率，分光光度法测定抗氧化指标丙二醛(MDA)，过氧化氢酶(CAT)，超氧化物歧化酶(SOD)和谷胱甘肽(GSH)，采用实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤坏死因子-

(TNF-

)，白细胞介素-1

(IL-1

)和白细胞介素-6(IL-6) mRNA表达水平，采用免疫荧光检测核转录因子-

B(NF-

B)p65的核移位情况。

结果：

浓度为250 μmol·L

-1

干预24 h后SH-SY5Y细胞存活率约55%为最佳造模条件，DSS高、中、低剂量干预后，与模型组比较，细胞活率呈剂量依赖性上升(

0.05)，MDA明显下调(

0.05)，抗氧化指标CAT，SOD和GSH明显上升(

0.05)；H

能显著上调SH-SY5Y细胞炎性因子TNF-

，IL-1

和IL-6 mRNA表达水平，并介导胞质NF-

B的激活和向核移位，当归芍药散含药血清能呈剂量依赖性抑制NF-

B p65的入核并降低IL-1

，IL-6和TNF-

等细胞炎性因子的mRNA表达水平。

结论：

当归芍药散含药血清可以显著降低H

诱导的SH-SY5Y细胞的氧化损伤通过改善其抗氧化状态，同时也能降低炎症反应通过抑制NF-

B信号通路。

Abstract

Objective:

To investigate the regulatory effect of Danggui Shaoyaosan (DSS)-containing serum on oxidative stress and inflammation in H

-induced SH-SY5Y cells.

Method:

Methyl thiazolyl tetrazolium(MTT) assay was used to determine the cell activity and construct the H

-induced cell damage model with the optimal time and dose. Normal group

model group and high

medium and low-dose DSS groups(2.5%

10%) were set up. MTT method was used to detect cell activity

spectrophotometry anti-oxidation indexes of malonaldehyde (MDA)

catalase (CAT) and superoxide dismutase (SOD) and glutathione (GSH). Real-time quantitative PCR (Real-time PCR) was used to detect tumor necrosis factor-

(TNF-

)

interleukin-1

(IL-1

) and interleukin-6 (IL-6) mRNA expressions. And immunofluorescence test was adopted to detect nuclear transcription factor-

B(NF-

B) p65 nuclear translocation of the DSS after the intervention.

Result:

After 24 h intervention with 250 μmol·L

-1

SH-SY5Y cell viability was about 55%

which was the best modeling condition. After high

medium and low-dose DSS intervention on H

-damaged cell model

compared with the model group

the cell activity showed a dose-dependent increase (

0.05)

MDA was significantly reduced (

0.05)

and antioxidant indexes CAT

SOD and GSH were significantly increased (

0.05). H

could significantly increase SH-SY5Y cell inflammatory factor TNF-

IL-6 and IL-1

mRNA expressions

and promote activation of cytoplasmic NF-

B and nuclear translocation. DSS-containing serum showed a dose-dependent inhibition of NF-

B p65 from nuclear

and reduced inflammatory factor levels

such as TNF-

IL-6 and IL-1

Conclusion:

DSS-containing serum can significantly reduce the oxidative damage in H

-induced SH-SY5Y cells by improving their antioxidant status

and reduce the inflammatory response by inhibiting NF-

B signaling pathway.

关键词

Keywords

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Effect of Danggui Shaoyaosan Ameliorates in Alleviating H2O2-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway

DOI：10.13422/j.cnki.syfjx.20200604

摘要

Abstract

关键词

Keywords

references

Effect of Danggui Shaoyaosan Ameliorates in Alleviating H₂O₂-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway