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姜黄素通过 miR-125a-5p/SIRT6 轴调节 ABCA1 表达促进胆固醇流出,从而防止动脉粥样硬化发生在 THP-1 巨噬细胞中。

Curcumin promotes cholesterol efflux by regulating ABCA1 expression through miR-125a-5p/SIRT6 axis in THP-1 macrophage to prevent atherosclerosis.

机构信息

Department of Internal Medicine, the First Affiliated Hospital of Hunan University of Chinese Medicine, China.

The Domestic First-class Discipline Construction Project of Chinese Medicine of Hunan University of Chinese Medicine, China.

出版信息

J Toxicol Sci. 2021;46(5):209-222. doi: 10.2131/jts.46.209.

Abstract

OBJECTIVE

To seek out the effect of curcumin on cholesterol efflux in THP-1 macrophages and clarify its specific mechanism.

METHODS

THP-1 macrophages were cultured with curcumin at different concentrations, followed by detection of the toxicity of curcumin to cells utilizing CCK-8 assay. Following culturing with serum-free ox-LDL, THP-1 macrophages were transfected with mi-miR-125a-5p, or in-miR-125a-5p, or pcDNA3.1-SIRT6, or si-SIRT6 for 24 hr, prior to treatment with curcumin at different concentrations. Oil red O staining was applied to examine the formation rate of foam cells, the kits were used for measuring intracellular lipid content of THP-1 macrophages, and the fluorescence detection kit for observing the cholesterol efflux rate. The expressions of miR-125a-5p, SIRT6, and ABCA1 were assayed by qRT-PCR and Western blot. ELISA was adopted to assess the contents of TNF-α, IL-6, and MCP-1. The interaction between miR-125a-5p and SIRT6 was evaluated by dual-luciferase reporter gene assay.

RESULTS

The optimal dosage of curcumin could reduce foam cell formation and intracellular lipid content, and promote cholesterol efflux in THP-1 macrophages. Meanwhile, curcumin markedly suppressed the expression of miR-125a-5p and upregulated the expression of SIRT6. MiR-125a-5p negatively targeted SIRT6. Overexpression of SIRT6 partially reversed the inhibition role of miR-125a-5p mimic in the biological function of curcumin. Silencing of SIRT6 could partially reverse the effect of the miR-125a-5p inhibitor on the biological function of curcumin.

CONCLUSION

urcumin could promote cholesterol efflux of THP-1 macrophages through miR-125a-5p/SIRT6 axis and regulate the expression of ABCA1.

摘要

目的

探讨姜黄素对 THP-1 巨噬细胞胆固醇外流的影响,并阐明其具体机制。

方法

用不同浓度的姜黄素培养 THP-1 巨噬细胞,用 CCK-8 法检测姜黄素对细胞的毒性。用无血清 ox-LDL 孵育 THP-1 巨噬细胞后,转染 mi-miR-125a-5p、in-miR-125a-5p、pcDNA3.1-SIRT6 或 si-SIRT6 24 小时,再用不同浓度的姜黄素处理。油红 O 染色检测泡沫细胞形成率,试剂盒检测 THP-1 巨噬细胞内脂质含量,荧光检测试剂盒观察胆固醇流出率。qRT-PCR 和 Western blot 检测 miR-125a-5p、SIRT6 和 ABCA1 的表达。ELISA 法检测 TNF-α、IL-6 和 MCP-1 的含量。双荧光素酶报告基因检测 miR-125a-5p 与 SIRT6 的相互作用。

结果

最佳剂量的姜黄素可减少 THP-1 巨噬细胞泡沫细胞形成和细胞内脂质含量,促进胆固醇外流。同时,姜黄素显著抑制 miR-125a-5p 的表达,上调 SIRT6 的表达。miR-125a-5p 负靶向 SIRT6。SIRT6 过表达部分逆转 miR-125a-5p 模拟物对姜黄素生物学功能的抑制作用。沉默 SIRT6 可部分逆转 miR-125a-5p 抑制剂对姜黄素生物学功能的影响。

结论

姜黄素通过 miR-125a-5p/SIRT6 轴促进 THP-1 巨噬细胞胆固醇外流,调节 ABCA1 的表达。

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