Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury

Wei HUANG; Han LI; Ben-yue LI; Yang ZHANG; Li GUAN; Qi WANG

doi:10.13422/j.cnki.syfjx.20191102

您当前的位置：

首页 >

文章列表页 >

Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury

Pharmacology | 更新时间：2021-02-09

- Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 11, Pages: 67-72(2019)
- 作者机构：
  
  1.广州中医药大学　临床药理研究所，广州　 510006；
  2.广州中医药大学　基础医学院，广州　 510006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191102
  CLC： R2-0; R22; R285.5
- Received：09 November 2018，
  
  Published Online：19 February 2019，
  
  Published：05 June 2019
- 稿件说明：
移动端阅览
Wei HUANG, Han LI, Ben-yue LI, et al. Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(11): 67-72.
DOI：

Wei HUANG, Han LI, Ben-yue LI, et al. Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(11): 67-72. DOI： 10.13422/j.cnki.syfjx.20191102.

摘要

目的：

研究川芎嗪(TMP)对叔丁基过氧化氢(t-BHP)诱导的PC12细胞损伤的保护作用及对磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/雷帕霉素靶蛋白(mTOR)信号通路的影响。

方法：

培养PC12细胞，孵育不同浓度的川芎嗪(10，25，50，100，200 μmol·L

－1

)，12 h后采用细胞增殖毒性检测试剂盒(CCK-8)确定川芎嗪的保护浓度。随后将 PC12 细胞分为空白组，t-BHP 组，TMP组。空白组加入培养基，t-BHP组加入t-BHP(200 μmol·L

－1

)，TMP组加入TMP(25，50，100 μmol·L

－1

)预处理 12 h 后加入t-BHP (200 μmol·L

－1

)。共作用6 h后采用酶联免疫吸附测定(ELISA)检测乳酸脱氢酶(LDH)漏出量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、活性氧(ROS)含量和谷胱甘肽过氧化物酶(GSH-Px)活性。采用蛋白免疫印迹法(Western blot)检测B淋巴细胞瘤-2(Bcl-2)，Bcl-2相关X蛋白(Bax)，Akt，磷酸化Akt(p-Akt)及mTOR，p-mTOR的表达情况。

结果：

与空白组比较，t-BHP组细胞生存率降低(

0.01)，与t-BHP组比较，25，50，100 μmol·L

－1

的川芎嗪促进细胞增殖(

0.05，

0.01)，其中50 μmol·L

－1

组的细胞增殖率最高(

0.01)；与空白组比较，t-BHP组LDH的释放量，ROS和MDA含量显著增加(

0.01)，SOD和GSH-Px的活性显著降低(

0.01)，与t-BHP组比较，TMP组LDH的释放量，ROS和MDA含量显著降低(

0.05，

0.01)，SOD和GSH-Px的活性显著升高(

0.05，

0.01)；Western blot表明与空白组比较，t-BHP组中p-Akt，p-mTOR蛋白表达水平明显下降(

0.01)，Bcl-2/Bax值明显降低(

0.01)，与t-BHP组比较，TMP(50 μmol·L

－1

)组中p-Akt，p-mTOR蛋白表达水平明显上升(

0.05)，Bcl-2/Bax值明显上升(

0.05)。另外LY294002(PI3K蛋白抑制剂)处理减弱了这些变化。

结论：

川芎嗪通过激活PI3K/Akt/mTOR信号通路保护t-BHP诱导的PC12细胞损伤。

Abstract

Objective:

To study the protective effect of tetramethylpyrazine (TMP) on PC12 cells induced by tert-butyl hydroperoxide (t-BHP) and the regulatory mechanism on signaling pathway of phosphatidylinositol-3-kinases (PI3K)/kinase B (Akt)/mammalian target of rapamycin(mTOR).

Method:

PC12 cells cultured

in vitro

were treated with t-BHP (200 μmol·L

-1

) for 6 h to establish a model of oxidative damage in PC12 cells. The experiment was divided into blank group

model group (200 μmol·L

-1

t-BHP)

TMP group. PC12 cells were pretreated with TMP(25

100 μmol·L

-1

) for 12 h

and then treated with t-BHP for 6 h. The cell viability was detected by cell counting kit-8(CCK-8) method

and lactate dehydrogenase (LDH) leakage

malondialdehyde (MDA) content

superoxide dismutase (SOD) activity

reactive oxygen species (ROS) and glutathione peroxidase (GSH-Px) activity were detected by enzyme-linked immunosorbent assay (ELISA). Apoptosis was observed by annexin V-FITC/PI double staining. B-cell lymphoma-2 (Bcl-2)

Bcl-2-associated X protein (Bax)

total protein kinase B (Akt)

and phosphorylated protein kinase B (p-Akt)

mTOR and p-mTOR expressions were detected by Western blot.

Result:

The cell viability of PC12 cells treated with 200 μmol·L

-1

t-BHP decreased to about 50%after 6 h. This condition was suitable for the establishment of oxidative damage model. Compared with the model group

TMP (25

100 μmol·L

-1

) pretreatment for 12 h significantly increased the survival rate of PC12 cells (

0.05

0.01)

decreased the release of LDH and MDA (

0.05

0.01)

significantly reduced the ROS content in PC12 cells (

0.01)

significantly increased the activity of SOD

GSH-Px (

0.05

0.01)

and inhibited the cell apoptosis. Western blot showed that compared with the model group

the protein expressions of p-Akt and p-mTOR in TMP (50 μmol·L

-1

) pretreatment group increased significantly (

0.01)

and the Bcl-2/Bax ratio increased significantly. Treatment with LY294002 (PI3K protein inhibitor) also attenuated these changes.

Conclusion:

Ligustrazine protects PC12 cell injury induced by t-BHP by activating PI3K/Akt/mTOR signaling pathway.

关键词

Keywords

references

CHEN Z C , ZHONG C J . Oxidative stress in Alzheimer's disease [J]. Neurosci Bull , 2014 , 30 ( 2 ): 271 - 281 .

Sandra F I , Miquel V , Celine P . Mitochondrial quality control in neurodegenerative diseases: focus on parkinson's disease and huntington's disease [J]. Front Neurosci , 2018 , 12 : 1 - 25 .

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

傅瑜，廖琴，樊东升．神经保护剂治疗缺血性脑卒中的研究进展 [J]．中国新药杂志， 2011 ， 20 ( 11 )： 973 - 977 ．

周惠芬，何昱，张宇燕，等．川芎和黄芪有效部位组合给药后川芎嗪在脑缺血再灌注大鼠体内的PK-PD结合研究 [J]．中草药， 2016 ， 47 ( 19 )： 3463 - 3468 ．

吴平勇，曾宪涛，王学军，等．川芎嗪对心肌缺血再灌注损伤防治作用的系统评价与Meta分析 [J]．中国循证心血管医学杂志， 2014 ， 6 ( 6 )： 653 - 657 ．

李健琳，韩江全，陈玲．川芎嗪和葛根素配伍对脑缺血大鼠神经细胞调亡及Caspase-3影响 [J]．齐齐哈尔医学院学报， 2016 ， 37 ( 4 )： 423 - 425 ．

林文新，马阮昕，冯真英．川芎嗪对过氧化氢诱导PC12细胞氧化应激的作用 [J]．江西中医药， 2018 ， 49 ( 2 )： 29 - 31 ．

李杰．川芎嗪联合血栓通治疗缺血性脑卒中的疗效分析 [J]．中国实用神经疾病杂志， 2014 ， 17 ( 17 )： 108 - 109 ．

刘文铠，肖辉亮，周徐州．丹参川芎嗪注射液辅助辨治缺血性脑卒中的临床研究 [J]．中国生化药物杂志， 2014 ， 34 ( 2 )： 120 - 121 ．

Ghosh M , Manna P , Sil P C . Protective role of a coumarin-derived schiff base scaffold against tertiary butyl hydroperoxide (TBHP)-induced oxidative impairment and cell death via MAPKs, NF-kappaB and mitochondria-dependent pathways [J]. Free Radic Res , 2011 , 45 ( 5 ): 620 - 637 .

Bajpai V K , Alam M B , Ju M K , et al . Antioxidant mechanism of polyphenol-rich Nymphaea nouchali leaf extract protecting DNA damage and attenuating oxidative stress-induced cell death via Nrf2-mediated heme-oxygenase-1 induction coupled with ERK/p38 signaling pathway [J]. Biomed Pharmacother , 2018 , 103 : 1397 - 1407 .

陈剑平，李中桂，张尚斌，等．大枣神经保护作用的活性组分筛选及其作用机制研究 [J]．中国药房， 2016 ． 27 ( 25 )： 3495 - 3498 ．

高婷，王子旭，陈祝茗，等． ROS介导的氧化应激与自噬 [J]．中国畜牧兽医， 2018 ， 45 ( 3 )： 656 - 662 ．

ZHOU J , MA X , CUI Y , et al . Methyleugenol protects against t-BHP-triggered oxidative injury by induction of Nrf2 dependent on AMPK/GSK3beta and ERK activation [J]. J Pharmacol Sci , 2017 , 135 ( 2 ): 55 - 63 .

SHEN C , MA W , ZHENG W , et al . The antioxidant effects of riluzole on the APRE-19 celll model injury-induced by t-BHP [J]. BMC Ophthalmol , 2017 , 17 ( 1 ): 210 .

高健美，李德芬，牛爽，等．金丝桃苷对过氧化氢诱导的A549细胞氧化损伤的保护作用 [J]．中国实验方剂学杂志， 2017 ， 23 ( 11 )： 128 - 133 ．

刘亭，宋菲，杨健，等．基于HSF1高表达H9c2细胞的参芎葡萄糖注射液抗氧化损伤作用探讨 [J]．中国实验方剂学杂志， 2018 ， 24 ( 8 )： 85 - 90 ．

张建武，闵冬雨，周云，等．番茄红素对H2O2致乳鼠心肌细胞氧化应激损伤的保护作用 [J]．中国实验方剂学杂志， 2014 ， 20 ( 12 )： 160 - 164 ．

林文新，马阮昕，冯真英．川芎嗪对过氧化氢诱导PC12细胞氧化应激的作用 [J]．江西中医药， 2018 ， 49 ( 2 )： 29 - 31 ．

Aydemir T , Oztürk R , Bozkaya L A , et al . Effects of antioxidant vitamins A, C, E and trace elements Cu, Se on CuZn SOD, GSH-Px, CAT and LPO levels in chicken erythrocytes [J]. Cell Biochem Funct , 2000 , 18 ( 2 ): 109 - 115 .

陈丹丹，彭成，李梦婷，等．自噬参与氢溴酸樟柳碱对缺氧状态下PC12细胞损伤的保护作用 [J]．中国实验方剂学杂志， 2017 ， 23 ( 20 )： 144 - 149 ．

卢春凤，杨玉，商宇，等． ROS介导的氧化应激在INH诱导的细胞毒性中的作用及槲皮素的干预 [J]．生态毒理学报， 2015 ， 10 ( 3 )： 209 - 215 ．

严钢莉，李朝武，聂海岭，等．丹参川芎嗪注射液对Aβ损伤的PC12细胞保护作用及机制研究 [J]．卒中与神经疾病， 2016 ， 23 ( 1 )： 15 - 20 ．

TANG F , ZHOU X , WANG L , et al . A novel compound DT-010 protects against doxorubicin-induced cardiotoxicity in zebrafish and H9c2 cells by inhibiting reactive oxygen species-mediated apoptotic and autophagic pathways [J]. Eur J Pharmacol , 2018 , 820 : 86 - 96 .

WANG L , ZHANG H Y , GAO B , et al . Tetramethylpyrazine protects against glucocorticoid-induced apoptosis by promoting autophagy in mesenchymal stem cells and improves bone mass in glucocorticoid-induced osteoporosis rats [J]. Stem Cells Dev , 2017 , 26 ( 6 ): 419 - 430 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Modified Buwangsan Ameliorates Cognitive Dysfunction in Rat Model of Type 2 Diabetes Mellitus by Regulating Autophagy in Hippocampus via PI3K/Akt/mTOR Pathway

Protective Effect and Mechanism of Anmeidan against Neuronal Damage in Rat Model of Sleep Deprivation Based on Hippocampal Neuroinflammation

Effect of Artemisinin and Its Derivatives in Treatment of Nervous System Diseases： A Review

Neuroprotective Effect of Ginsenoside Rb₁ on Lipopolysaccharide-induced Neuroinflammation in Mice Based on PI3K/Akt Signaling Pathway

Feasibility of Tetramethylpyrazine in Prevention and Treatment of Alzheimer's Disease Based on Theory of 'Toxin Damaging Brain Collaterals'

Related Author

YANG Jie

LIU Tonghua

LIU Wei

WU Lili

QIN Lingling

XIE Guangjing

XU Zixuan

ZHANG Junlu

Related Institution

Science and Technology Department， Beijing University of Chinese Medicine

Key Laboratory of Health Preservation and Rehabilitation， Ministry of Education， Beijing University of Chinese Medicine

Dongfang Hospital， Beijing University of Chinese Medicine

Hubei Provincial Hospital of Traditional Chinese Medicine

Hubei Shizhen Laboratory

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰