川芎嗪对过氧化氢诱导星形胶质细胞氧化应激损伤的保护作用

杨琴; 漆国栋; 伍亚民; 王海燕; 徐珂; 侯科名; 江琼

doi:10.16016/j.1000-5404.202008129

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川芎嗪对过氧化氢诱导星形胶质细胞氧化应激损伤的保护作用

神经科学 | Updated：2026-01-09

- 川芎嗪对过氧化氢诱导星形胶质细胞氧化应激损伤的保护作用
- Tetramethylpyrazine protects rat astrocytes against H₂O₂-induced oxidative stress injury
- 第三军医大学学报 2021年43卷第3期页码：226-233
- Affiliations：
  
  1.Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Metabolic Diseases, College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing, 400016
  2.Department of Special Environmental Warfare Injury Prevention and Control, Institute of Surgery Research, Army Characteristic Medical Center, Chongqing, 400042
  3.Department of Orthopedics, Chongqing Orthopedic Hospital of Traditional Chinese Medicine, Chongqing, 400010, China
- Author bio：
  
  [ 江琼，E-mail：101755@cqmu.edu.cn JIANG Qiong, E-mail: 101755@cqmu.edu.cn
- Funds：
  
  重庆市科卫联合中医药科研项目(2019ZY023495)
- DOI：10.16016/j.1000-5404.202008129
  中图分类号： R282.71;R285.5;R322.8
- 收稿：2020-08-17，
  
  修回：2020-09-23，
  
  纸质出版：2021-02-15
- Accepted：
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杨琴, 漆国栋, 伍亚民, 等. 川芎嗪对过氧化氢诱导星形胶质细胞氧化应激损伤的保护作用[J]. 第三军医大学学报, 2021,43(3):226-233.

Qin YANG, Guodong QI, Yamin WU, et al. Tetramethylpyrazine protects rat astrocytes against H₂O₂-induced oxidative stress injury[J]. Journal of Third Military Medical University, 2021, 43(3): 226-233.
杨琴, 漆国栋, 伍亚民, 等. 川芎嗪对过氧化氢诱导星形胶质细胞氧化应激损伤的保护作用[J]. 第三军医大学学报, 2021,43(3):226-233. DOI： 10.16016/j.1000-5404.202008129.

Qin YANG, Guodong QI, Yamin WU, et al. Tetramethylpyrazine protects rat astrocytes against H₂O₂-induced oxidative stress injury[J]. Journal of Third Military Medical University, 2021, 43(3): 226-233. DOI： 10.16016/j.1000-5404.202008129.

Abstract

目的探讨川芎嗪(tetramethylpyrazine，TMP)对过氧化氢(H₂O₂)所致大鼠星形胶质细胞凋亡的保护作用。方法利用SD新生鼠(24~48 h)分离、培养星形胶质细胞，按随机数字表法分为空白对照组(以正常培养液培养)、模型组(100 μmol/L的H₂O₂作用24 h)、TMP剂量组(25、50、100 μmol/L TMP预处理48 h后，加入H₂O₂作用24 h)。采用CCK-8检测各组细胞存活率；GFAP染色观察细胞形态；流式细胞术检测各组细胞凋亡率；特定试剂盒检测乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活力；Real-time PCR检测各组细胞中PI3K、AKT、Bcl-2、Bax、Caspase-3 mRNA的表达；Western blot检测各组细胞PI3K、AKT、p-AKT、Bcl-2、Bax、Caspase-3的蛋白表达。结果与空白对照组比较，模型组细胞存活率明显降低(P < 0.05)，细胞凋亡率升高(P < 0.05)，LDH含量增加(P < 0.05)，SOD活性降低(P < 0.05)，PI3K、AKT、Bcl-2 mRNA表达下调，Bax、Caspase-3 mRNA表达上调(P < 0.05)，PI3K、p-AKT、Bcl-2蛋白表达明显降低，Bax、Caspase-3蛋白表达明显升高(P < 0.05)；与模型组比较，TMP预处理后细胞存活率明显升高(P < 0.05)，其中50 μmol/L组的细胞存活率最高(P < 0.05)，细胞凋亡率明显降低(P < 0.05)，LDH含量降低(P < 0.05)，SOD活性升高(P < 0.05)，PI3K、AKT、Bcl-2 mRNA表达上调，Bax、Caspase-3 mRNA表达下调(P < 0.05)，PI3K、p-AKT、Bcl-2蛋白表达明显升高，Bax、Caspase-3蛋白表达明显降低(P < 0.05)。结论TMP对H₂O₂所致的大鼠星形胶质细胞氧化应激损伤具有保护作用，可改善损伤后细胞形态并抑制凋亡，该作用可能与PI3K、p-AKT蛋白活化有关。

Keywords

摘要

目的

探讨川芎嗪(tetramethylpyrazine，TMP)对过氧化氢(H

)所致大鼠星形胶质细胞凋亡的保护作用。

方法

利用SD新生鼠(24~48 h)分离、培养星形胶质细胞，按随机数字表法分为空白对照组(以正常培养液培养)、模型组(100 μmol/L的H

作用24 h)、TMP剂量组(25、50、100 μmol/L TMP预处理48 h后，加入H

作用24 h)。采用CCK-8检测各组细胞存活率；GFAP染色观察细胞形态；流式细胞术检测各组细胞凋亡率；特定试剂盒检测乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活力；Real-time PCR检测各组细胞中PI3K、AKT、Bcl-2、Bax、Caspase-3 mRNA的表达；Western blot检测各组细胞PI3K、AKT、p-AKT、Bcl-2、Bax、Caspase-3的蛋白表达。

结果

与空白对照组比较，模型组细胞存活率明显降低(

0.05)，细胞凋亡率升高(

0.05)，LDH含量增加(

0.05)，SOD活性降低(

0.05)，PI3K、AKT、Bcl-2 mRNA表达下调，Bax、Caspase-3 mRNA表达上调(

0.05)，PI3K、p-AKT、Bcl-2蛋白表达明显降低，Bax、Caspase-3蛋白表达明显升高(

0.05)；与模型组比较，TMP预处理后细胞存活率明显升高(

0.05)，其中50 μmol/L组的细胞存活率最高(

0.05)，细胞凋亡率明显降低(

0.05)，LDH含量降低(

0.05)，SOD活性升高(

0.05)，PI3K、AKT、Bcl-2 mRNA表达上调，Bax、Caspase-3 mRNA表达下调(

0.05)，PI3K、p-AKT、Bcl-2蛋白表达明显升高，Bax、Caspase-3蛋白表达明显降低(

0.05)。

结论

TMP对H

所致的大鼠星形胶质细胞氧化应激损伤具有保护作用，可改善损伤后细胞形态并抑制凋亡，该作用可能与PI3K、p-AKT蛋白活化有关。

Abstract

Objective

To investigate the protective effect of tetramethylprazine (TMP) on oxidative stress injur

y induced by hydrogen peroxide (H

) in rat astrocytes.

Methods

Astrocytes were isolated from SD newborn rats (24~48 h old) and cultured

and then the cells were randomly divided into control group (cultured with normal culture medium)

model group (100 μmol·L

-1

for 24 h)

TMP groups (pretreatment of 25

100 μmol·L

-1

TMP for 48 h

and then H

was added for 24 h). CCK-8 assay was used to detect cell survival rate

GFAP staining to observe cell morphology

flow cytometry to detect cell apoptosis

corresponding reagent test kits to detect the lactate dehydrogenase (LDH) content and superoxide dismutase (SOD) activity

real-time fluorescent quantitative PCR to measure the detect the mRNA expression of PI3K

AKT

Bcl-2

Bax and Caspase-3

and Western blotting to test the protein expression levels of PI3K

AKT

p-AKT

Bcl-2

Bax and Caspase-3 in each group of cells.

Results

The model group had significantly reduced cell survival rate (

0.05)

increased apoptotic rate (

0.05)

elevated LDH content (

0.05)

decreased SOD activity (

0.05); down-regulated PI3K

AKT

Bcl-2 mRNA and protein levels

and up-regulated the expression levels of Bax and Caspase-3 mRNA and protein levels (

0.05). Compared with the model group

TMP pretreatment resulted in significantly increased cell survival rate (

0.05)

lowest apoptotic rate (

0.05) and LDH content (

0.05)

enhanced SOD activity (

0.05)

up-regulation of PI3K

AKT

and Bcl-2 at mRNA and protein levels

and down-regulation of Bax and Caspase-3 at mRNA and protein levels (

0.05).

Conclusion

TMP exerts protective effects on oxidative stress injury of rat astrocytes i

nduced by H

and improves cell morphology and inhibits cell apoptosis after injury

and this effect may be related to the regulation of PI3K and p-AKT activation.

关键词

Keywords

references

MATIAS I, MORGADO J, GOMES F C A. Astrocyte heterogeneity: impact to brain aging and disease[J]. Front Aging Neurosci, 2019, 11: 59. DOI: 10.3389/fnagi.2019.00059.

CARTER S F, HERHOLZ K, ROSA-NETO P, et al . Astrocyte biomarkers in Alzheimer's disease[J ] . Trends Mol Med, 2019, 25(2): 77-95. DOI: 10.1016/j.molmed.2018.11.006.

BOISON D, STEINHÄUSER C. Epilepsy and astrocyte energy metabolism[J]. Glia, 2018, 66(6): 1235-1243. DOI: 10.1002/glia.23247.

CHEN L, LIU B. Relationships between stress granules, oxidative stress, and neurodegenerative diseases[J]. Oxid Med Cell Longev, 2017, 2017: 1809592. DOI: 10.1155/2017/1809592.

CABEZAS R, BAEZ-JURADO E, HIDALGO-LANUSSA O, et al . Growth factors and neuroglobin in astrocyte protection against neurodegeneration and oxidative stress[J ] . Mol Neurobiol, 2019, 56(4): 2339-2351. DOI: 10.1007/s12035-018-1203-9.

GUO M, LIU Y, SHI D. Cardiovascular actions and therapeutic potential oftetramethylpyrazine (active component isolated from rhizoma Chuanxiong): roles and mechanisms[J]. Biomed Res Int, 2016, 2016: 2430329. DOI: 10.1155/2016/2430329.

谢保城, 陈世春, 王清辉, 等.丹参川芎嗪注射液辅助治疗急性脑梗死疗效与安全性的系统评价[J].中国中药杂志, 2018, 43(17): 3573-3581. DOI: 10.19540/j.cnki.cjcmm. 20180611.015.

XIE B C, CHEN S C, WANG Q H, et al .Systematic review on efficacy and safety of Danshen Chuanxiongqin Injection in treatment of acute cerebral infarction[J ] . China J Chin Materia Medica, 2018, 43(17): 3573-3581. DOI: 10.19540/j.cnki.cjcmm. 20180611.015.

ZHANG G X, ZHANG T, WU L M. et al .Neuruprotective effect and mechanism of action of tetramethylpyrazine nitrone for ischemic stroke therapy[J ] . Neuromolecular Med, 2018, 20(1): 97-111. DOI: 10.1007/s12017-018-8478-x.

喻志慧, 黎良达.川芎嗪对小鼠脑缺血再灌注损伤的影响[J].中国临床药理学杂志, 2020, 36(12): 1648-1652. DOI:10.13699/j.cnki.1001-6821.2020.12.013.

YU Z H, LI L D. Effect of ligustrazine on the cerebral ischemia-reperfusion injury in mice[J]. Chin J Clin Pharmacol, 2020, 36(12): 1648-1652.DOI: 10.13699/j.cnki.1001-6821.2020.12.013.

JIANG T, SUN Q, CHEN S. Oxidative stress: a major pathogenesis and potential therapeutic target of antioxidative agents in Parkinson's disease and Alzheimer's disease[J]. Prog Neurobiol, 2016, 147: 1-19. DOI: 10.1016/j.pneurobio.2016.07.005.

CHO C H, KIM E A, KIM J, et al . N-Adamantyl-4-methylthiazol-2-amine suppresses amyloid β-induced neuronal oxidative damage in cortical neurons[J ] . Free Radic Res, 2016, 50(6): 678-690. DOI: 10.3109/10715762.2016.1167277.

陈艳清, 甄然, 杨璇, 等.星形胶质细胞的功能作用研究进展[J].脑与神经疾病杂志, 2020, 28(5): 327-330.

CHEN Y Q, ZHEN R, YANG X, et al . Research progress on the function of astrocytes[J ] . J Brain Nerv Dis, 2020, 28(5): 327-330.

孙旋, 冯卓蕾, 陈娜.川芎嗪对过氧化氢致大鼠视网膜神经节细胞氧化损伤的抑制作用及机制[J].眼科新进展, 2019, 39(6): 515-518. DOI: 10.13389/j.cnki.rao.2019.0118.

SUN X, FENG Z L, CHEN N. Inhibitory effect of ligustrazine on rat retinal ganglion cell oxidative damage induced by peroxide hydrogen and its mechanisms[J]. Rec Adv Ophthalmol, 2019, 39(6): 515-518. DOI: 10.13389/j.cnki.rao.2019.0118.

黄伟, 李涵, 李本岳, 等.川芎嗪对t-BHP致PC12细胞损伤的保护及机制[J].中国实验方剂学杂志, 2019, 25(11): 67-72. DOI: 10.13422/j.cnki.syfjx.20191102.

HUANG W, LI H, LI B Y, et al . Protective effect and mechanism of tetramethylpyrazine against t-BHP-induced PC12 cell injury[J ] . Chin J Exper Trad Med Formul, 2019, 25(11): 67-72. DOI: 10.13422/j.cnki.syfjx.20191102.

GUAN D S, SU Y F, LI Y X, et al . Tetramethylpyrazine inhibits CoCl 2 -induced neurotoxicity through enhancement of Nrf2/GCLc/GSH and suppression of HIF1α/NOX2/ROS pathways[J ] . J Neurochem, 2015, 134(3): 551-565. DOI: 10.1111/jnc.13161..

QUINCOZES-SANTOS A, BOBERMIN L D, LATINI A, et al . Resveratrol protects C6 astrocyte cell line against hydrogen peroxide-induced oxidative stress through heme oxygenase 1[J ] . PLoS ONE, 2013, 8(5): e64372. DOI: 10.1371/journal.pone.0064372.

黑鑫鑫, 刘涛.石菖蒲活性成分对过氧化氢诱导PC12细胞Caspase-1、IL-18表达的影响[J].时珍国医国药, 2019, 30(5): 1025-1028.

HEI X X, LIU T. Protective effects of β-asarone, an active ingredient of Acorus tatarinowii, on the pyroptosis of PC12 c ells induced by H 2 O 2 [J ] . Lishizhen Med Mater Med Res, 2019, 30(5): 1025-1028..

李尽文, 许月莹, 曹天寿, 等.姜黄素对过氧化氢诱导PC12细胞氧化应激损伤的保护作用[J].四川化工, 2019, 22(3): 1-5.

LI J W, XU Y Y,CAO T S, et al . The neuroprotective effects of curcumin against H 2 O 2 -induced oxidative stress in PC12 cells[J ] . Sichuan Chem Ind, 2019, 22(3): 1-5..

D'ARCY M S. Cell death: a review of the major forms of apoptosis, necrosis and autophagy[J]. Cell Biol Int, 2019, 43(6): 582-592. DOI: 10.1002/cbin.11137.

ZHAO W W, FENG H T, SUN W, et al . Tert-butyl hydroperoxide (t-BHP) induced apoptosis and necroptosis in endothelial cells: Roles of NOX4 and mitochondrion[J ] . Redox Biol, 2017, 11: 524-534. DOI: 10.1016/j.redox.2016.12.036.

SÁNCHEZ-MIGALLÓN M C, VALIENTE-SORIANO F J, NADAL-NICOLÁS F M, et al . Apoptotic retinal ganglion cell death after optic nerve transection or crush in mice: delayed RGC loss with BDNF or a caspase 3 inhibitor[J ] . Invest Ophthalmol Vis Sci, 2016, 57(1): 81. DOI: 10.1167/iovs.15-17841.

YU J S, CUI W. Proliferation, survival and metabolism: the role of PI 3 K/AKT/mTOR signalling in pluripotency and cell fate determination[J ] . Development, 2016, 143(17): 3050-3060. DOI: 10.1242/dev.137075.

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