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Effects and Mechanisms of Autophagy Induced by Solubilized-Cholesterol in Hepatocytes: A Comparative Study Among Solvents

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Abstract

Cholesterol, the principal sterol in mammalian cells, has been reported to play a role in the pathogenesis of several diseases through autophagy. Due to its insoluble characteristic, all in vitro cholesterol experiments are performed using dimethyl sulphoxide, methyl-β-cyclodextrin, and ethanol co-solvents. To investigate whether the types of solvents have different effects on cholesterol-induced cell behaviors, we analyzed the effects and mechanisms of autophagy induced by solubilized-cholesterol in hepatic cells. We found that both solubilized-cholesterol and involved solvents could induce autophagy. Solubilized-cholesterol could further enhance the LC3-II expression with or without the pre-treatment with lysosomal blockers compared with the single-solvent groups, indicating that cholesterol could sensitize cells to solvents-induced autophagy. Besides, solubilized-cholesterol and single-solvent treatment could repress the activation of AKT-mTOR pathway. Furthermore, cholesterol solubilized in methyl-β-cyclodextrin could induce apoptosis while other solubilized-cholesterol or single solvent groups could not, suggesting that different dissolve methods may affect the cytotoxic of cholesterol. These results strongly suggest that the effect of solvent should be taken into consideration in further in vitro cholesterol studies.

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Acknowledgements

The subject was supported by: The National Natural Science Foundation of China (No. 81370952).

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Author notes

  1. These authors contributed equally: Dong Liang, Ezra K. Osoro

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi’an Jiaotong University Health Science Center, Xi’an, China

    Dong Liang, Ezra K. Osoro, Xi Lan, Wenhua Zhu, Litao Wu, Xiaojuan Du, Dongmin Li & Shemin Lu

  2. Key Laboratory of Environment and Genes Related to Diseases, Xi’an Jiaotong University, Ministry of Education of China, Beijing, China

    Dong Liang, Ezra K. Osoro, Xi Lan, Wenhua Zhu, Litao Wu, Xiaojuan Du, Dongmin Li & Shemin Lu

  3. Department of Medicine, Solna, Karolinska Universitetssjukhuset, Stockholm, Sweden

    Shuai Tan

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Liang, D., Osoro, E.K., Tan, S. et al. Effects and Mechanisms of Autophagy Induced by Solubilized-Cholesterol in Hepatocytes: A Comparative Study Among Solvents. Cell Biochem Biophys 78, 357–366 (2020). https://doi.org/10.1007/s12013-020-00917-2

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