Beijing University of Chinese Medicine, Beijing, China (mainland).
Central Laboratory, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing, China (mainland).
Med Sci Monit. 2019 Feb 21;25:1373-1382. doi: 10.12659/MSM.915123.
BACKGROUND Autophagy is characterized by the degradation of cellular components in autophagosomes. It plays a significant role in cerebral ischemic injury and has a complex functional connection with apoptosis. The neurovascular unit (NVU) is a structural and functional unit of the nervous system presented as a therapeutic target of stroke. This study aimed to investigate the effect of autophagy induced by ischemic damage on NVUs. MATERIAL AND METHODS SH-SY5Y cells, C6 cells, and rat brain microvascular endothelial cells were cultured with oxygen-glucose deprivation (OGD) exposure for different time durations, and 3-methyladenine (3-MA) was added as an autophagy inhibitor. In all 3 cell lines, lactate dehydrogenase (LDH) release was measured. Furthermore, apoptosis was detected using Annexin V-fluorescein isothiocyanate/propidium iodide labeling and immunofluorescence staining. Autophagosomes were observed through AO/MDC (acridine orange/monodansycadaverine) double staining. LC3-II expression levels were evaluated by western blot analysis. RESULTS In the OGD groups of 3 cell lines, LDH leakage, and apoptotic rates were obviously increased. Remarkable increase in LC3-II expression was found in the OGD groups of SH-SY5Y cells and C6 cells. However, 3-MA decreased the LC3-II expression to varying degrees. CONCLUSIONS OGD could induce the over-activation of autophagy and augment the apoptotic activity in neurons and glial cells of NVUs.
自噬的特征是在自噬体中降解细胞成分。它在脑缺血损伤中起重要作用，与细胞凋亡有复杂的功能联系。神经血管单元（NVU）是神经系统的结构和功能单位，是中风的治疗靶点。本研究旨在探讨缺血损伤诱导的自噬对 NVU 的影响。
用氧葡萄糖剥夺（OGD）暴露培养 SH-SY5Y 细胞、C6 细胞和大鼠脑微血管内皮细胞不同时间，并加入 3-甲基腺嘌呤（3-MA）作为自噬抑制剂。在所有 3 种细胞系中，测量乳酸脱氢酶（LDH）释放。此外，通过 Annexin V-荧光素异硫氰酸酯/碘化丙啶标记和免疫荧光染色检测细胞凋亡。通过吖啶橙/单丹磺酰尸胺（AO/MDC）双重染色观察自噬体。通过 Western blot 分析评估 LC3-II 表达水平。
在 3 种细胞系的 OGD 组中，LDH 漏出和凋亡率明显增加。在 SH-SY5Y 细胞和 C6 细胞的 OGD 组中，LC3-II 表达明显增加。然而，3-MA 不同程度地降低了 LC3-II 的表达。
OGD 可诱导 NVU 神经元和神经胶质细胞中自噬的过度激活，并增加细胞凋亡活性。
