玛咖水提物在PC12细胞对抗化学性缺氧损伤中作用

张静; 陈萌; 赵君; 周雯; 李慧; 唐慧; 张天亮; 范轶欧

doi:10.11847/zgggws1119716

玛咖水提物在PC12细胞对抗化学性缺氧损伤中作用

Effect of maca water extract on cobalt chloride-induced hypoxia injury in PC12 cells

摘要

摘要:
目的探讨玛咖水提物在PC12细胞对抗二氯化钴（CoCl₂）诱导化学性缺氧损伤中的作用及机制。
方法应用不同浓度的CoCl₂处理PC12细胞不同时间，建立化学性缺氧损伤模型。应用细胞增殖检验试剂盒（CCK-8）检测细胞存活率；试剂盒法测定细胞培养液中超氧化物歧化酶（SOD）和一氧化氮合酶（NOS）的活性，检测丙二醛、一氧化氮（NO）及乳酸脱氢酶（LDH）的含量；NO荧光探针（DAF-FM DA）染色流式细胞术检测细胞内NO含量；Annexin V-FITC/PI 染色检测细胞凋亡率。
结果与对照组比较，CoCl₂组PC12细胞存活率明显受到抑制，且呈现一定的浓度与时间依赖性；与对照组比较，300 μmol/L CoCl₂组PC12细胞存活率（51.55 ± 2.71）%明显降低，细胞培养液中SOD、NOS活性升高，丙二醛、NO和LDH含量增加；与CoCl₂组比较，60、120 μg/mL玛咖组PC12细胞存活率分别为（63.99 ± 3.24）%、（69.95 ± 2.25）%明显升高，细胞培养液中NOS活性降低，丙二醛、NO和LDH含量降低。与对照组比较，500 μmol/L CoCl₂组PC12细胞内NO含量（61.5 ± 5.7） %明显升高，细胞凋亡率（41.33 ± 3.83）%明显增加；与500 μmol/L CoCl₂组比较，60、120 μg/mL玛咖组细胞内NO含量分别为（36.5 ± 4.7）%、（33.4 ± 3.8）%明显降低，细胞凋亡率分别为（34.87 ± 2.50）%、（31.33 ± 3.44）%明显下降。
结论玛咖水提物对CoCl₂诱导的化学性缺氧损伤具有一定拮抗作用，其机制可能与抑制NO的产生、减轻氧化应激反应，抑制细胞凋亡有关。

Abstract:
Objective To explore the effect of maca water extract on cobalt chloride (CoCl₂)-induced hypoxia injury in PC12 cells.
Methods PC12 cells were exposed to CoCl₂ at different dose and for different time duration to set up the chemical hypoxia injury model. Cell viability was determined with cell counter kit-8 (CCK-8). The activity of superoxide dismutase (SOD) and nitric oxide synthase (NOS), the quantity of malondialdehyde (MDA), nitric oxide (NO), and lactate dehydrogenase (LDH) in culture medium were measured with kit assay. The intracellular NO was detected using a fluorescent probe (2′,7′-dichlorodihydrofluorescein diacetate, DCFH-DA). The cell apoptotic rate was determined with annexin V-FITC/PI staining.
Results Compared with that of the control cells, the viability of PC12 cells exposed to CoCl₂ was inhibited significantly in a dose- and time-dependent manner. Significantly decreased cell viability rate (51.55% ± 2.71%) and increased of SOD and NOS and content of MDA, NO, and LDH in culture medium were observed in PC12 cells exposed to 300 μmol/L CoCl₂ for 24 hours in comparison with those of the control cells. For the PC12 cells treated with maca water extraction of 60 and 120 μg/ml, the viability rate increased obviously (63.99% ± 3.24% and 69.95% ± 2.25%) and the NOS activity and content of MDA, NO, and LDH in culture medium decreased compared to the CoCl₂ exposed PC12 cells. Obviously increased intracellular NO content (61.5% ± 5.7%) and cell apoptotic rate (41.33% ± 3.83%) were observed in the PC12 cells treated with 500 μmol/L CoCl₂ for 24 hours compared to the control cells. Obviously decreased intracellular NO content (36.50% ± 4.70% and 33.40% ± 3.80%) and cell apoptotic rate (34.87 ± 2.50% and 31.33% ± 3.44%) were observed in the PC12 cells treated with maca water extraction of 60 and 120 μg/ml compared to those in the cells exposed to 500 μmol/L CoCl₂.
Conclusion Maca water extract has protective effect on cobalt chloride-induced hypoxia injury by inhibiting NO generation, oxidative stress and cell apoptosis in PC12 cells.

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