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白藜芦醇通过下调缺氧诱导因子-1α减少小鼠RAW264.7巨噬细胞凋亡

[Resveratrol reduces apoptosis of mouse RAW264.7 macrophage via down-regulating HIF-1α].

作者信息

Zheng Haijun, Qiu Cuiting, Jin Hui, Sun Yachao, Li Zhongdong

机构信息

Department of Cardiology, People's Hospital of Jiaozuo City, Jiaozuo 454000, China.

Department of Hematology, People's Hospital of Jiaozuo City, Jiaozuo 454000, China. *Corresponding author, E-mail:

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Jan;36(1):42-48.

Abstract

Objective To investigate the protective effect of resveratrol (Res) against mouse RAW264.7 macrophage injury induced by cobalt chloride (CoCl) and its mechanism. Methods Macrophages were divided into control group, CoCl group and Res pretreatment group. CoCl group was treated with 500 μmol/L CoCl for 8 hours, and Res pretreatment group was pretreated with 40 μmol/L Res for 2 hours followed by 500 μmol/L CoCl treatment for 8 hours. The cell vitality and apoptotic rate in every group were detected by CCK-8 assay and flow cytometry. The distributions of caspase-3 and hypoxia-inducible factor 1 alpha (HIF-1α), as well as the influences of CoCl and Res on their expression were detected by immunofluorescence cytochemistry. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in every group were measured by the corresponding kits. The expression levels of Bcl2, BAX, c-caspase-3 and HIF-1α in every group were observed by Western blot analysis. Results Compared with the CoCl group, pretreatment with Res increased cell vitality, decreased apoptosis, enhanced the activity of SOD, and reduced the level of MDA. The caspase-3 was mainly distributed in the cytoplasm, and HIF-1α was mainly distributed on the nucleus. Compared with the CoCl group, Res up-regulated the expression of Bcl2 and down-regulated the expression of BAX, cleaved caspase-3 and HIF-1α. Conclusion Res can decrease apoptosis in macrophages, which may occur via reducing the accumulation of intracellular reactive oxygen species, enhancing cell antioxidant capacity, and down-regulating the expression of HIF-1α on cells.

摘要

目的 探讨白藜芦醇(Res)对氯化钴(CoCl)诱导的小鼠RAW264.7巨噬细胞损伤的保护作用及其机制。方法 将巨噬细胞分为对照组、CoCl组和Res预处理组。CoCl组用500 μmol/L CoCl处理8小时,Res预处理组先用40 μmol/L Res预处理2小时,随后用500 μmol/L CoCl处理8小时。采用CCK-8法和流式细胞术检测各组细胞活力和凋亡率。通过免疫荧光细胞化学检测半胱天冬酶-3(caspase-3)和缺氧诱导因子1α(HIF-1α)的分布以及CoCl和Res对其表达的影响。用相应试剂盒检测各组超氧化物歧化酶(SOD)和丙二醛(MDA)水平。通过蛋白质免疫印迹分析观察各组Bcl2、BAX、c-caspase-3和HIF-1α的表达水平。结果 与CoCl组相比,Res预处理可提高细胞活力、降低凋亡率、增强SOD活性并降低MDA水平。caspase-3主要分布于细胞质,HIF-1α主要分布于细胞核。与CoCl组相比,Res上调Bcl2表达,下调BAX、裂解的caspase-3和HIF-1α的表达。结论 Res可减少巨噬细胞凋亡,其机制可能是通过减少细胞内活性氧的积累、增强细胞抗氧化能力以及下调细胞上HIF-1α的表达来实现。

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