低压缺氧比常压缺氧更能增加PC12大鼠嗜铬细胞瘤细胞的自噬和凋亡。

IF 1.6 4区 医学 Q4 BIOPHYSICS
Jiaojiao Yin, Yuhang Wang, Bing Li, Xiaoyan Hu, Yao Ma, Chong Zhang, Xiaoqin Ha, Linyan Wang, Yaozhu Pan
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引用次数: 0

摘要

尹娇娇,王玉航,李冰,胡小燕,马瑶,张冲,哈小琴,王林燕,潘耀柱。低压缺氧比常压缺氧更能促进PC12大鼠嗜铬细胞瘤细胞的自噬和凋亡。高Alt医学生物杂志,200,2025。目的:目前,体外研究主要集中在急性高山病(AMS)的缺氧损伤,但对低气压缺氧的影响评价较少。AMS是一种神经系统疾病,大鼠嗜铬细胞瘤(PC12)细胞是研究神经元存活和凋亡的模型。本研究开发了一种模拟高空低压缺氧的细胞培养方法,比较了低压缺氧和常压缺氧对PC12细胞自噬和凋亡的影响。方法:将PC12细胞分别培养于正常、常压缺氧、低压缺氧条件下。透射电镜和免疫荧光显微镜观察细胞自噬现象。Western blot检测缺氧诱导因子1-α (HIF1-α)、LC3、caspase-3、cleaved caspase-3的表达水平。结果:模拟高空低气压缺氧的细胞培养室,气压保持在41.1 kPa,氧密度保持在1% (PO2约3.08 mmHg)。低压缺氧处理PC12细胞0、4、8、16、24和48 h时,HIF1-α和LC3Ⅱ蛋白水平升高,HIF1-α/actin和LC3Ⅱ/actin比值均在16 h达到峰值(p < 0.05)。0.01),细胞存活率为88.02%。与常压缺氧16小时的PC12细胞相比,低压缺氧时LC3Ⅱ表达增加1.5倍,lc3b阳性点增加2倍,自噬体积累增加(p < 0.001)。有趣的是,在常压缺氧条件下,促进PC12细胞自噬(与雷帕霉素或3-MA共培养)可降低裂解型caspase-3的表达(裂解型caspase-3/caspase-3的比值降低,p < 0.01)。然而,在低气压低氧条件下,自噬促进作用使裂解型caspase-3呈反比升高(裂解型caspase-3/caspase-3比值升高,p < 0.01),而自噬抑制作用(羟氯喹[HCQ]共培养)使裂解型caspase-3降低(裂解型caspase-3/caspase-3比值降低,p < 0.01)。结论:与常压缺氧细胞相比,体外培养的低压缺氧细胞自噬和凋亡增加。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Hypobaric Hypoxia Increased Autophagy and Apoptosis in PC12 Rat Pheochromocytoma Cells More Than Normobaric Hypoxia.

Jiaojiao Yin, Yuhang Wang, Bing Li, Xiaoyan Hu, Yao Ma, Chong Zhang, Xiaoqin Ha, Linyan Wang, and Yaozhu Pan. Hypobaric hypoxia increased autophagy and apoptosis in PC12 rat pheochromocytoma cells more than normobaric hypoxia. High Alt Med Biol. 00:00-00, 2025. Purpose: Currently, in vitro studies have focused on hypoxia injury in acute mountain sickness (AMS), but little effort has been made to assess the effects of hypobaric hypoxia. AMS is a neurological disorder, and rat pheochromocytoma (PC12) cells are a model to study neuronal survival and apoptosis. Here, we developed a novel cell culture method that mimics hypobaric hypoxia at high attitude and compared the effects of hypobaric hypoxia and normobaric hypoxia on autophagy and apoptosis of PC12 cells. Methods: PC12 cells were cultured under normal conditions, normobaric hypoxia, and hypobaric hypoxia. Autophagy was observed by transmission electron microscopy and immunofluorescence microscopy. The hypoxia-inducible factor1-α (HIF1-α), LC3, caspase-3, and cleaved caspase-3 expression levels were determined by Western blot. Results: The cell culture chamber mimicking hypobaric hypoxia at high attitude perfectly maintained the air pressure at 41.1 kPa and the oxygen density at 1% (PO2 around 3.08 mmHg). Hypobaric hypoxic treatment of PC12 cells at 0, 4, 8, 16, 24, and 48 hours resulted in an increase in HIF1-α and LC3Ⅱ protein levels, and the ratio of HIF1-α/actin and LC3Ⅱ/actin both peaked at 16 hours (p < 0. 01) when the cell viability was 88.02%. There was a 1.5-fold increase in LC3Ⅱ expression, a 2-fold increase in LC3B-positive spots, and an increase in autophagosome accumulation at hypobaric hypoxia compared to PC12 cells at normobaric hypoxia for 16 hours (p < 0.001). Interestingly, the promotion of autophagy (coculture with rapamycin or 3-MA) in PC12 cells under normobaric hypoxia reduced cleaved caspase-3 expression (the ratio of cleaved caspase-3/caspase-3 decreased, p < 0.01). However, under hypobaric hypoxia, the promotion of autophagy inversely increased cleaved caspase-3 (the ratio of cleaved caspase-3/caspase-3 increased, p < 0.01), and the inhibition of autophagy (hydroxychloroquine [HCQ] coculture) decreased cleaved caspase-3 (the ratio of cleaved caspase-3/caspase-3 decreased, p < 0.01). Conclusions: Compared with normobaric hypoxia cells, hypobaric hypoxia cells cultured in vitro exhibited increased autophagy and apoptosis.

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来源期刊
High altitude medicine & biology
High altitude medicine & biology 医学-公共卫生、环境卫生与职业卫生
CiteScore
3.80
自引率
9.50%
发文量
44
审稿时长
>12 weeks
期刊介绍: High Altitude Medicine & Biology is the only peer-reviewed journal covering the medical and biological issues that impact human life at high altitudes. The Journal delivers critical findings on the impact of high altitude on lung and heart disease, appetite and weight loss, pulmonary and cerebral edema, hypertension, dehydration, infertility, and other diseases. It covers the full spectrum of high altitude life sciences from pathology to human and animal ecology.
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