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Anti-apoptotic Effects of Probucol are Associated with Downregulation of Daxx Expression in THP-1 Macrophage

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Abstract

Aim

To study the relationship between Daxx expression and the antiapoptotic effects of probucol in THP-1 macrophage.

Materials and methods

Apoptosis of THP-1 derived macrophages was induced by exposure to oxidized low density lipoprotein (oxLDL). The development of apoptosis was determined by flow cytometry analysis and nucleic acid-binding dye acridin orange. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and indirect immunofluorescence were used to evaluate the expression of Daxx and caspase-3 at both mRNA and protein level.

Results

As expected, THP-1 macrophages exposed to 100 mg/l oxLDL for 48 h exhibited typical morphologic changes of apoptosis, including condensed chromatin and shrunken nucleus. oxLDL treatment markedly increased Daxx expression in a time- and dose-dependent manner, and facilitated Daxx translocation from cytoplasm to nucleus. The percentage of cells with Daxx in nuclei was significantly increased from 8 to 59%. Treatment with probucol (50 μmol/l) for 4 h prior to exposure to oxLDL significantly inhibited Daxx expression and THP-1 macrophage apoptosis by 61.3%. Furthermore, oxLDL enhanced caspase-3 expression with increased mRNA and protein levels, but without obvious change in translocation of caspase-3 (the cells with nuclear Daxx: 14 vs 8%). In contrast, probucol attenuated oxLDL-stimulated caspase-3 expression in THP-1 macrophages.

Conclusion

OxLDL-induced apoptosis of THP-1 macrophage is associated with Daxx up-regulation; while inhibition of apoptosis by probucol is related to decreased Daxx expression and nuclear translocation.

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Acknowledgments

The authors are indebted to Dr. Li-Xin Li (Oregon University, USA), Dr. He-Sheng Ou (Baylor College of Medicine, USA) and Dr. Zhen-Gao Liu (Ohio State University, USA) for their critical discussions, suggestions, and valuable help in the preparation of the manuscript. This work was supported by the National Natural Science Foundation of China (30470719, 30600249), the National Major Basic Research Program of China (973) (2006CB503808), and the Education Department of Hunan Province (99B11).

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Authors and Affiliations

  1. Division of Pharmacoproteomics, Institute of Pharmacy & Pharmacology, Nanhua University, Changsheng Western Road 28#, Hengyang, 421001, Hunan, China

    Bo Su, Yun-Bo Yang, Qin-Hui Tuo, Bing-Yang Zhu, Xiao-Yong Lei & Duan-Fang Liao

  2. Department of Biochemistry and Molecular Biology, School of Life Science and Technology, Nanhua University, Changsheng Western Road 28#, Hengyang, 421001, Hunan, China

    Weidong Yin & Duan-Fang Liao

  3. Department of Pharmacology, Central South University, Changsha, Hunan, 410078, China

    Yun-Bo Yang, Qin-Hui Tuo & Duan-Fang Liao

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  1. Bo Su
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Correspondence to Duan-Fang Liao.

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Bo Su and Yun-Bo Yang equally contributed to this paper.

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Su, B., Yang, YB., Tuo, QH. et al. Anti-apoptotic Effects of Probucol are Associated with Downregulation of Daxx Expression in THP-1 Macrophage. Cardiovasc Drugs Ther 21, 37–45 (2007). https://doi.org/10.1007/s10557-007-6002-x

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