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当归芍药散通过调控NF-
κ B炎性通路改善H2O2诱导的SH-SY5Y细胞氧化损伤的作用Effect of Danggui Shaoyaosan Ameliorates in Alleviating H2O2-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-
κ B Pathway- 中国实验方剂学杂志 2020年26卷第10期 页码：1-7
- 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：
[第一作者] 余婧萍，硕士，从事中西医结合神经退行性疾病防治研究，E-mail：838619403@qq.com
*宋祯彦，实验师，从事中西医结合神经退行性疾病防治研究，E-mail：songzhenyan2013@hnucm.edu.cn
- 基金信息：国家自然科学基金面上项目(81774129);湖南省自然科学基金面上项目(2018JJ2296;2019JJ50441);湖南省中医药管理局科研项目(2018025;201609);湖南省教育厅科研项目(15K088;18B246);2018年湖南中医药大学大学生研究性学习和创新性实验计划课题项目(2018017)
DOI：10.13422/j.cnki.syfjx.20200604
中图分类号： R2-0; R22; R285.5; R289收稿日期：2019-09-04，
网络出版日期：2019-12-06，
纸质出版日期：2020-05-20
- 稿件说明：
移动端阅览
余婧萍, 贺春香, 成绍武, 等. 当归芍药散通过调控NF-
κ B炎性通路改善H2O2诱导的SH-SY5Y细胞氧化损伤的作用[J]. 中国实验方剂学杂志, 2020,26(10):1-7.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-
κ B Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 1-7.余婧萍, 贺春香, 成绍武, 等. 当归芍药散通过调控NF-
κ B炎性通路改善H2O2诱导的SH-SY5Y细胞氧化损伤的作用[J]. 中国实验方剂学杂志, 2020,26(10):1-7. DOI： 10.13422/j.cnki.syfjx.20200604.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-
κ B 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：
2
研究当归芍药散(Danggui Shaoyaosan，DSS)含药血清对过氧化氢(H
2
O
2
)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：
2
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
2
O
2
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-
α
(TNF-
α
)，白细胞介素-1
β
(IL-1
β
)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-
κ
B(NF-
κ
B)p65的核移位情况。
结果：
2
H
2
O
2
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(
P
<
0.05)，MDA明显下调(
P
<
0.05)，抗氧化指标CAT，SOD和GSH明显上升(
P
<
0.05)；H
2
O
2
能显著上调SH-SY5Y细胞炎性因子TNF-
α
，IL-1
β
和IL-6 mRNA表达水平，并介导胞质NF-
κ
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-
κ
B p65的入核并降低IL-1
β
，IL-6和TNF-
α
等细胞炎性因子的mRNA表达水平。
结论：
2
当归芍药散含药血清可以显著降低H
2
O
2
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-
κ
B信号通路。
Abstract
Objective:
2
To investigate the regulatory effect of Danggui Shaoyaosan (DSS)-containing serum on oxidative stress and inflammation in H
2
O
2
-induced SH-SY5Y cells.
Method:
2
Methyl thiazolyl tetrazolium(MTT) assay was used to determine the cell activity and construct the H
2
O
2
-induced cell damage model with the optimal time and dose. Normal group
model group and high
medium and low-dose DSS groups(2.5%
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-
α
(TNF-
α
)
interleukin-1
β
(IL-1
β
) and interleukin-6 (IL-6) mRNA expressions. And immunofluorescence test was adopted to detect nuclear transcription factor-
κ
B(NF-
κ
B) p65 nuclear translocation of the DSS after the intervention.
Result:
2
After 24 h intervention with 250 μmol·L
-1
H
2
O
2
SH-SY5Y cell viability was about 55%
which was the best modeling condition. After high
medium and low-dose DSS intervention on H
2
O
2
-damaged cell model
compared with the model group
the cell activity showed a dose-dependent increase (
P
<
0.05)
MDA was significantly reduced (
P
<
0.05)
and antioxidant indexes CAT
SOD and GSH were significantly increased (
P
<
0.05). H
2
O
2
could significantly increase SH-SY5Y cell inflammatory factor TNF-
α
IL-6 and IL-1
β
mRNA expressions
and promote activation of cytoplasmic NF-
κ
B and nuclear translocation. DSS-containing serum showed a dose-dependent inhibition of NF-
κ
B p65 from nuclear
and reduced inflammatory factor levels
such as TNF-
α
IL-6 and IL-1
β
.
Conclusion:
2
DSS-containing serum can significantly reduce the oxidative damage in H
2
O
2
-induced SH-SY5Y cells by improving their antioxidant status
and reduce the inflammatory response by inhibiting NF-
κ
B signaling pathway.
关键词
Keywords
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