Medical College, Jiaxing University, Jiaxing, 314001, China.
Institute of Traditional She Medicine, Department of Pharmacy, Lishui People's Hospital, Lishui, 323000, China.
J Ethnopharmacol. 2020 May 10;253:112614. doi: 10.1016/j.jep.2020.112614. Epub 2020 Jan 31.
Harpagide is the main ingredient in Scrophularia ningpoensis Hemsl which is used for the therapeutic purpose of treating encephalopathy. Harpagide has shown promise in the treatment of oxygen-glucose deprivation and reoxygenation (OGD/R)-induced brain injury. However, the underlying mechanisms remain unclear.
In this study, we aimed to determine the neuroprotective effect of harpagide on rat cortical neurons under OGD/R conditions that induce the development of ischaemia-reperfusion (I/R).
To explore the biological function of harpagide in cerebral ischaemia-reperfusion injury (CIRI), The CIRI model was established by oxygen-glucose deprivation and reoxygenation (OGD/R) on rat cortical neurons. It tested cell survival rate by 3-(4,5-dimethylthiazol-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, apoptosis by flow cytometry, intracellular Ca concentration [Ca] by cofocal laser, and expressions related to endoplasmic reticulum stress (ERS) by RT-PCR and Western blot.
We found that pretreatment with harpagide (50 μM) prevented OGD/R-induced apoptotic cell death. Harpagide also significantly decreased the gene expression levels and protein production of ERS-related proteins. We found that harpagide also exerted a neuroprotective effect on TG-induced apoptosis in rat cortical neurons and decreased the gene expression levels and protein production of GRP78, caspase-12 and CHOP. We also measured the intracellular calcium ion concentration ([Ca]) in neurons and found that harpagide significantly decreased the [Ca] induced by OGD/R and TG.
These results suggest that harpagide protects against OGD/R-induced cell apoptosis, likely by decreasing ERS. Collectively, harpagide was demonstrated to be a prominent suppressor of ERS and prevented the apoptosis of rat cortical neurons. Based on the results, harpagide could potentially serve as a therapeutic agent of ischaemia-like injury associated with excessive ERS and apoptosis.
哈巴苷是玄参 Scrophularia ningpoensis Hemsl 的主要成分，用于治疗脑病。哈巴苷在治疗氧葡萄糖剥夺和再氧合（OGD/R）诱导的脑损伤方面显示出良好的效果。然而，其潜在机制尚不清楚。
本研究旨在确定哈巴苷在氧葡萄糖剥夺和再氧合（OGD/R）诱导的缺血再灌注（I/R）条件下对大鼠皮质神经元的神经保护作用。
为了探讨哈巴苷在脑缺血再灌注损伤（CIRI）中的生物学功能，采用氧葡萄糖剥夺和再氧合（OGD/R）建立大鼠皮质神经元 CIRI 模型。通过 3-(4,5-二甲基噻唑-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）比色法检测细胞存活率，通过流式细胞术检测细胞凋亡，通过共聚焦激光检测细胞内 Ca 浓度 [Ca]，通过 RT-PCR 和 Western blot 检测与内质网应激（ERS）相关的表达。
我们发现，哈巴苷（50 μM）预处理可预防 OGD/R 诱导的凋亡性细胞死亡。哈巴苷还显著降低了 ERS 相关蛋白的基因表达水平和蛋白产物。我们发现哈巴苷还对 TG 诱导的大鼠皮质神经元凋亡具有神经保护作用，并降低了 GRP78、caspase-12 和 CHOP 的基因表达水平和蛋白产物。我们还测量了神经元内钙离子浓度 ([Ca])，发现哈巴苷显著降低了 OGD/R 和 TG 诱导的 [Ca]。
这些结果表明，哈巴苷通过降低 ERS 来保护神经元免受 OGD/R 诱导的细胞凋亡。综上所述，哈巴苷被证明是 ERS 的显著抑制剂，并可防止大鼠皮质神经元凋亡。基于这些结果，哈巴苷可能成为与 ERS 过度和细胞凋亡相关的缺血样损伤的治疗药物。
