Mechanism of pterostilbene protecting HT22 cells from hydrogen peroxide-induced apoptosis

FAN Chong-xi; ZHANG Yun-shui; SUN Bing-bing; WANG Xiao-ying; ZHAO Dong-lin; YANG Quan-long; SHI Xue-hui; LIU Lei

doi:10.11659/jjssx.04E023029

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Mechanism of pterostilbene protecting HT22 cells from hydrogen peroxide-induced apoptosis

Basic Research | 更新时间：2023-10-23

- Mechanism of pterostilbene protecting HT22 cells from hydrogen peroxide-induced apoptosis
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 32, Issue 10, Pages: 839-844(2023)
- 作者机构：
  
  1.空军特色医学中心消化内科，北京 100142
  2.空军特色医学中心重症医学科，北京 100142
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.04E023029
  CLC： R741.05
- Published：25 October 2023，
  
  Received：29 April 2023，
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范崇熙,张云水,孙兵兵等.紫檀芪保护HT22细胞抗过氧化氢诱导凋亡的机制研究[J].局解手术学杂志,2023,32(10):839-844.

FAN Chong-xi,ZHANG Yun-shui,SUN Bing-bing,et al.Mechanism of pterostilbene protecting HT22 cells from hydrogen peroxide-induced apoptosis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(10):839-844.
范崇熙,张云水,孙兵兵等.紫檀芪保护HT22细胞抗过氧化氢诱导凋亡的机制研究[J].局解手术学杂志,2023,32(10):839-844. DOI： 10.11659/jjssx.04E023029.

FAN Chong-xi,ZHANG Yun-shui,SUN Bing-bing,et al.Mechanism of pterostilbene protecting HT22 cells from hydrogen peroxide-induced apoptosis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(10):839-844. DOI： 10.11659/jjssx.04E023029.

摘要

目的

探讨紫檀芪（PTE）对过氧化氢（H

）诱导HT22细胞氧化应激损伤的保护作用及调控机制。

方法

不同浓度的PTE处理HT22细胞12 h，并构建H

氧化应激损伤模型。CCK-8法检测细胞活力，比色法检测细胞乳酸脱氢酶（LDH）释放量，TUNEL法染色检测细胞凋亡，DCFH-DA测定活性氧（ROS）水平，JC-1标记线粒体膜电位（MMP），并通过Western blot检测内质网应激和凋亡相关蛋白表达情况。

结果

CCK-8结果显示，PTE浓度小于10.0 μmol/L时，HT22细胞活性无明显变化。H

可显著抑制HT22细胞活性，5.0 μmol/L和10.0 μmol/L PTE可有效增加损伤后细胞活性，减少LDH释放量，抑制凋亡及相关蛋白的表达，同时减少细胞内ROS，稳定MMP和细胞色素C分布。Western blot检测发现，PTE降低H

损伤后PERK和eIF2α的磷酸化水平以及下调Bip、CHOP的表达。

结论

PTE通过维持线粒体稳态、抑制内质网应激，进而减轻氧化应激诱导的细胞凋亡，最终发挥保护HT22细胞抗H

损伤的作用。

Abstract

Objective

To investigate the protective effect and regulatory mechanism of pterostilbene （PTE） on hydrogen peroxide （H

）-induced oxidative stress injury in HT22 cells.

Methods

HT22 cells were treated with different concentrations of PTE for 12 hours，and the H

oxidative stress injury model was established. The cell viability was detected by CCK-8 assay. The release of lactate dehydrogenase （LDH） was evaluated by colorimetry. The apoptosis was detected by TUNEL staining method. The reactive oxygen species （ROS） level was detected by DCFH-DA，and the mitochondrial membrane potential （MMP） was marked by JC-1. Western blot was used to detect the expression of endoplasmic reticulum stress and apoptosis-related proteins.

Results

The results of CCK-8 showed that the activity of HT22 cells was not significantly changed when the concentration of PTE was less than 10.0 μmol/L. H

could significantly inhibit the activity of HT22 cells， 5.0 μmol/L and 10.0 μmol/L PTE could effectively increase the activity of injured cells，reduce the release of LDH， inhibit the expression of apoptosis and related proteins，and reduce intracellular ROS， stabilize MMP and the distribution of cytochrome C. Western blot showed that PTE decreased the phosphorylations level of PERK and eIF2α，and down-regulated the expression of Bip and CHOP after H

injury.

Conclusion

PTE protects HT22 cells against H

injury through stabilizing mitochondrial homeostasis，inhibiting endoplasmic reticulum stress，and then reducing oxidative stress-induced apoptosis.

关键词

紫檀芪HT22细胞内质网应激线粒体稳态氧化应激

Keywords

pterostilbeneHT22 cellsendoplasmic reticulum stressmitochondrial homeostasisoxidative stress

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