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Abstract

Biology

LPS和ATP诱导的PMA分化THP-1巨噬细胞死亡及其验证

Published: May 3rd, 2024

DOI:

10.3791/66831

1State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, 2Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine

Abstract

细胞死亡是所有生物体的基本过程。该方案建立了脂多糖 (LPS) 和三磷酸腺苷 (ATP) 诱导的佛波醇-12-肉豆蔻酸酯-13-乙酸酯 (PMA) 分化的人单核细胞 (THP-1) 巨噬细胞中的脂质沉积模型,以观察细胞死亡。LPS联合ATP是一种经典的炎症诱导方法,常用于研究焦亡,但细胞凋亡和坏死性凋亡也对LPS/ATP的刺激有反应。在正常情况下,磷脂酰丝氨酸仅局限于质膜的内小叶中。然而,在细胞焦亡、凋亡和坏死性凋亡的早期阶段,细胞膜保持完整并暴露于磷脂酰丝氨酸,而在后期阶段,细胞膜失去其完整性。本文采用流式细胞术分析膜联蛋白V和7-氨基放线菌素D(AAD)双染色,检测全细胞的细胞死亡情况。结果显示,在用LPS/ATP刺激后,大量细胞死亡。使用扫描电子显微镜,我们观察了单个细胞中细胞死亡的可能形式。结果表明,在用LPS/ATP刺激后,细胞可能会发生焦亡、凋亡或坏死性凋亡。该协议的重点是观察用LPS / ATP刺激后巨噬细胞的死亡。结果表明,LPS和ATP刺激后的细胞死亡不仅限于焦亡,还可以发生凋亡和坏死性细胞凋亡,有助于研究人员更好地了解LPS和ATP刺激后的细胞死亡,并选择更好的实验方法。

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