Protective Effect of Dihuang Yinzi on Energy Deprivation-induced Endoplasmic Reticulum Stress in SH-SY5Y Cells

GAO Jun-feng; ZHANG Zhi-chen; WEN Bin-yu; TANG Xu; HUANG Qian-qian; MA Tao

doi:10.13422/j.cnki.syfjx.20182140

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Protective Effect of Dihuang Yinzi on Energy Deprivation-induced Endoplasmic Reticulum Stress in SH-SY5Y Cells

更新时间：2024-09-23

- Protective Effect of Dihuang Yinzi on Energy Deprivation-induced Endoplasmic Reticulum Stress in SH-SY5Y Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 21, Pages: 118-123(2018)
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- DOI：10.13422/j.cnki.syfjx.20182140
  CLC： R285.5
- Published：2018
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GAO Jun-feng, ZHANG Zhi-chen, WEN Bin-yu, et al. Protective Effect of Dihuang Yinzi on Energy Deprivation-induced Endoplasmic Reticulum Stress in SH-SY5Y Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 118-123.
DOI：

GAO Jun-feng, ZHANG Zhi-chen, WEN Bin-yu, et al. Protective Effect of Dihuang Yinzi on Energy Deprivation-induced Endoplasmic Reticulum Stress in SH-SY5Y Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 118-123. DOI： 10.13422/j.cnki.syfjx.20182140.

摘要

目的：以葡萄糖剥夺致能量代谢障碍SH-SY5Y细胞为模型，探讨地黄饮子药物血清通过调节蛋白激酶R样内质网激酶（protein kinase r-like endoplasmic reticulum kinase，PERK）/真核细胞起始因子2α（eukaryotic initiation factor 2α， eIF2α）通路改善能量代谢障碍致SH-SY5Y细胞内质网应激的作用机制。方法：以成人临床用药量10倍剂量对SD大鼠灌胃给药，制备地黄饮子药物血清。SH-SY5Y细胞葡萄糖剥夺处理6 h后，分别以0，1.25%，2.5%，5%，10%地黄饮子含药血清孵育。噻唑蓝（MTT）比色法检测细胞存活率，蛋白免疫印迹法（Western blot）检测PERK，eIF2α磷酸化水平，B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2），Bcl-2相关X蛋白（Bcl-2-associated X protein，Bax）表达水平。Annexin V/碘化丙啶（PI）双染流式细胞术检测细胞凋亡率。结果：与模型组比较，地黄饮子药物血清能显著提高葡萄糖剥夺SH-SY5Y细胞的存活率（P<0.01），明显减少PERK和eIF2α磷酸化激活（P<0.05，P<0.01），提高Bcl-2水平和降低Bax表达量（P<0.05，P<0.01）。抑制能量代谢障碍SH-SY5Y细胞凋亡率（P<0.01）。结论：地黄饮子可以减少PERK/eIF2α通路的激活，抑制能量代谢障碍致内质网应激造成的细胞凋亡。

Abstract

Objective: To investigate the effect and mechanism of Dihuang Yinzi-medicated serum on endoplasmic reticulum stress induced by glucose deprivation in SH-SY5Y cells. Method: SD rats were intragastrically administrated with Dihuang Yinzi at dosage of 10 times of the adult clinical dose to prepare Dihuang Yinzi-medicated serum. SH-SY5Y cells were deprivated by glucose for 6 h and then incubated with medium containing Dihuang Yinzi-medicated serum at concentrations of 0

1.25%

2.50%

5.00% and 10.00%

respectively. The cell viability was detected by methythiazol tetrazolium (MTT). The phosphorylation of protein kinase R-like endoplasmic reticulum kinase (PERK) and eukaryotic initiation factor 2α (eIF2α)

B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) expression levels were detected by Western blot. Annexin V/propidium iodide(PI) double staining flow cytometry were performed to detect the apoptotic rate. Result: As compared with the model group

Dihuang Yinzi-medicated serum could significantly increase the survival rate of SH-SY5Y cells deprived of glucose (P<0.01)

significantly reduce the phosphorylation of PERK and eIF2α (P<0.05

P<0.01)

increase the level of Bcl-2 and decrease the expression of Bax (P<0.05

P<0.01). Dihuang Yinzi-medicated serum could also inhibit apoptosis (P<0.05

P<0.01) induced by energy-deprivation in SH-SY5Y cells. Conclusion: Dihuang Yinzi-medicated serum can reduce the activation of PERK/eIF2α pathway and inhibit the apoptosis caused by energy deprivation-induced endoplasmic reticulum stress.

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