Bio‑IT Fusion Technology Research Institute, Pusan National University, Busan 46241, Republic of Korea.
Department of Horticultural Bioscience, Pusan National University, Miryang, Gyeongsangnam 50463, Republic of Korea.
Mol Med Rep. 2020 Jul;22(1):239-246. doi: 10.3892/mmr.2020.11075. Epub 2020 Apr 16.
Neuronal injury is a common, and critical, occurrence in clinical ischemic strokes, and can cause irreversible brain damage. However, the precise pathological mechanisms underlying this condition and effective treatment remain unclear. Increasing evidence shows that the nuclear factor erythroid 2‑related factor 2 (Nrf2)/activated protein kinase (AMPK) signaling pathway serves a significant role in neuronal injury and is involved in neuroprotection. The present study demonstrated that petatewalide B, the active constituent of Petasites japonicus, otherwise known as butterbur, can alleviate oxygen‑glucose deprivation/reoxygenation (OGD/R)‑induced neuronal death via the adenosine monophosphate‑AMPK/glycogen synthase kinase (GSK)‑3/β/Nrf2/antioxidant response element (ARE) signaling pathways in human neuroblastoma SH‑SY5Y cells. A neuronal injury model was established by depriving SH‑SY5Y cells of oxygen and glucose for 8 h, followed by 24 h of reoxygenation (OGD/R). The results indicated that the OGD/R model exhibited reduced cell viability but increased lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) production and apoptosis. These were accompanied by increased levels of cleaved PARP, cleaved caspase‑9, cleaved caspase‑3, p53, Bax and p21, as well as decreased Bcl‑2 levels. Treatment with petatewalide B was able to strengthen cell viability but reduced LDH release, ROS production and the expression levels of apoptosis‑related proteins. Additionally, treatment with petatewalide B activated AMPK in the OGD/R‑exposed SH‑SY5Y cells and upregulated activation of the downstream transcription factor Nrf2, which accompanied heme oxygenase 1 (HO‑1) and NAD(P)H quinone dehydrogenase 1 (NQO1) expression. Furthermore, silencing AMPK, Nrf2, HO‑1 and NQO1 expression inhibited petatewalide B's protective effect against apoptosis in the OGD/R‑exposed SH‑SY5Y cells. Therefore, petatewalide B protected human neuroblastoma cells against OGD/R‑induced injury by downregulating apoptosis and oxidative stress via upregulation of the AMPK/Nrf2 signaling pathway, suggesting that petatewalide B may be a prospective protector against neuronal injury, having possible therapeutic and medical implications.
神经元损伤是临床缺血性中风中常见且关键的事件，并可导致不可逆转的脑损伤。然而，这种情况的确切病理机制和有效治疗方法仍不清楚。越来越多的证据表明，核因子红细胞 2 相关因子 2（Nrf2）/蛋白激酶（AMPK）信号通路在神经元损伤中起重要作用，并参与神经保护。本研究表明，白头翁素 B，白头翁的活性成分，也称为蜂斗菜，可通过人神经母细胞瘤 SH-SY5Y 细胞中的腺苷一磷酸-AMPK/糖原合成酶激酶（GSK）-3/β/Nrf2/抗氧化反应元件（ARE）信号通路减轻氧葡萄糖剥夺/复氧（OGD/R）诱导的神经元死亡。通过剥夺 SH-SY5Y 细胞 8 h 的氧气和葡萄糖，然后进行 24 h 的复氧（OGD/R）建立神经元损伤模型。结果表明，OGD/R 模型表现出降低的细胞活力，但增加了乳酸脱氢酶（LDH）释放、活性氧（ROS）产生和细胞凋亡。这伴随着 PARP 裂解、caspase-9 裂解、caspase-3 裂解、p53、Bax 和 p21 水平的增加，以及 Bcl-2 水平的降低。用白头翁素 B 处理能够增强细胞活力，减少 LDH 释放、ROS 产生和细胞凋亡相关蛋白的表达水平。此外，在 OGD/R 暴露的 SH-SY5Y 细胞中，白头翁素 B 处理激活了 AMPK，并上调了下游转录因子 Nrf2 的激活，伴随着血红素加氧酶 1（HO-1）和 NAD（P）H 醌氧化还原酶 1（NQO1）的表达。此外，沉默 AMPK、Nrf2、HO-1 和 NQO1 表达抑制了白头翁素 B 对 OGD/R 暴露的 SH-SY5Y 细胞中细胞凋亡的保护作用。因此，白头翁素 B 通过下调凋亡和氧化应激来保护人神经母细胞瘤细胞免受 OGD/R 诱导的损伤，通过上调 AMPK/Nrf2 信号通路，提示白头翁素 B 可能是一种有前途的神经元损伤保护剂，具有潜在的治疗和医学意义。
