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The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages

PURPOSE: To investigate the sonoactivity of hypericin (HY), together with its sonodynamic effect on THP-1 macrophages and the underlying mechanism. MATERIALS AND METHODS: CCK-8 was used to examine cell viability. Confocal laser scanning microscopy was performed to assess the localization of HY in ce...

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Autores principales: Li, Xuesong, Gao, Lei, Zheng, Longbin, Kou, Jiayuan, Zhu, Xing, Jiang, Yueqing, Zhong, Zhaoyu, Dan, Juhua, Xu, Haobo, Yang, Yang, Li, Hong, Shi, Sa, Cao, Wenwu, Zhao, Yajun, Tian, Ye, Yang, Liming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309797/
https://www.ncbi.nlm.nih.gov/pubmed/25653524
http://dx.doi.org/10.2147/IJN.S75398
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author Li, Xuesong
Gao, Lei
Zheng, Longbin
Kou, Jiayuan
Zhu, Xing
Jiang, Yueqing
Zhong, Zhaoyu
Dan, Juhua
Xu, Haobo
Yang, Yang
Li, Hong
Shi, Sa
Cao, Wenwu
Zhao, Yajun
Tian, Ye
Yang, Liming
author_facet Li, Xuesong
Gao, Lei
Zheng, Longbin
Kou, Jiayuan
Zhu, Xing
Jiang, Yueqing
Zhong, Zhaoyu
Dan, Juhua
Xu, Haobo
Yang, Yang
Li, Hong
Shi, Sa
Cao, Wenwu
Zhao, Yajun
Tian, Ye
Yang, Liming
author_sort Li, Xuesong
collection PubMed
description PURPOSE: To investigate the sonoactivity of hypericin (HY), together with its sonodynamic effect on THP-1 macrophages and the underlying mechanism. MATERIALS AND METHODS: CCK-8 was used to examine cell viability. Confocal laser scanning microscopy was performed to assess the localization of HY in cells, reactive oxygen species (ROS) generation, and opening of the mitochondrial permeability transition pore (mPTP) after different treatments. Apoptosis was analyzed using Hoechst–propidium iodide and transmission electron microscopy. Mitochondrial membrane potential (ΔΨm) collapse was detected via fluorescence microscopy. Lipoprotein oxidation was determined in malondialdehyde (MDA) assays. Western blotting was conducted to determine the translocation of BAX and cytochrome C and the expression of apoptosis-related proteins. RESULTS: HY was sublocalized among the nuclei and the mitochondria, endoplasmic reticulum, Golgi apparatus, and lysosome in the cytosol of THP-1 macrophages. Under low-intensity ultrasound irradiation, HY significantly decreased cell viability and induced apoptosis. Furthermore, greater ROS generation, higher MDA levels, and greater ΔΨm loss were observed in the sonodynamic therapy (SDT) group. Both ROS generation and MDA levels were significantly reduced by the ROS scavenger N-acetyl cysteine (NAC) and the singlet oxygen scavenger sodium azide. Most of the loss of ΔΨm was inhibited by pretreatment with NAC, sodium azide, and the mPTP inhibitor cyclosporin A (CsA). mPTP opening was induced upon SDT but was reduced by pretreatment with bongkrekic acid, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid disodium, CsA, and NAC. Western blot analyses revealed translocation of BAX and cytochrome C, downregulated expression of Bcl-2, and upregulated expression of cleaved caspase-9, cleaved caspase-3, and cleaved poly(ADP-ribose) polymerase in the SDT group, which were reversed by NAC. CONCLUSION: HY mediated SDT-induced apoptosis in THP-1 macrophages via ROS generation. Then, the proapoptotic factor BAX translocated from the cytosol to the mitochondria, increasing the ratio of BAX/Bcl-2, and the mPTP opened to release cytochrome C. This study demonstrated the great potential of HY-mediated SDT for treating atherosclerosis.
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spelling pubmed-43097972015-02-04 The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages Li, Xuesong Gao, Lei Zheng, Longbin Kou, Jiayuan Zhu, Xing Jiang, Yueqing Zhong, Zhaoyu Dan, Juhua Xu, Haobo Yang, Yang Li, Hong Shi, Sa Cao, Wenwu Zhao, Yajun Tian, Ye Yang, Liming Int J Nanomedicine Original Research PURPOSE: To investigate the sonoactivity of hypericin (HY), together with its sonodynamic effect on THP-1 macrophages and the underlying mechanism. MATERIALS AND METHODS: CCK-8 was used to examine cell viability. Confocal laser scanning microscopy was performed to assess the localization of HY in cells, reactive oxygen species (ROS) generation, and opening of the mitochondrial permeability transition pore (mPTP) after different treatments. Apoptosis was analyzed using Hoechst–propidium iodide and transmission electron microscopy. Mitochondrial membrane potential (ΔΨm) collapse was detected via fluorescence microscopy. Lipoprotein oxidation was determined in malondialdehyde (MDA) assays. Western blotting was conducted to determine the translocation of BAX and cytochrome C and the expression of apoptosis-related proteins. RESULTS: HY was sublocalized among the nuclei and the mitochondria, endoplasmic reticulum, Golgi apparatus, and lysosome in the cytosol of THP-1 macrophages. Under low-intensity ultrasound irradiation, HY significantly decreased cell viability and induced apoptosis. Furthermore, greater ROS generation, higher MDA levels, and greater ΔΨm loss were observed in the sonodynamic therapy (SDT) group. Both ROS generation and MDA levels were significantly reduced by the ROS scavenger N-acetyl cysteine (NAC) and the singlet oxygen scavenger sodium azide. Most of the loss of ΔΨm was inhibited by pretreatment with NAC, sodium azide, and the mPTP inhibitor cyclosporin A (CsA). mPTP opening was induced upon SDT but was reduced by pretreatment with bongkrekic acid, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid disodium, CsA, and NAC. Western blot analyses revealed translocation of BAX and cytochrome C, downregulated expression of Bcl-2, and upregulated expression of cleaved caspase-9, cleaved caspase-3, and cleaved poly(ADP-ribose) polymerase in the SDT group, which were reversed by NAC. CONCLUSION: HY mediated SDT-induced apoptosis in THP-1 macrophages via ROS generation. Then, the proapoptotic factor BAX translocated from the cytosol to the mitochondria, increasing the ratio of BAX/Bcl-2, and the mPTP opened to release cytochrome C. This study demonstrated the great potential of HY-mediated SDT for treating atherosclerosis. Dove Medical Press 2015-01-22 /pmc/articles/PMC4309797/ /pubmed/25653524 http://dx.doi.org/10.2147/IJN.S75398 Text en © 2015 Li et al. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0) License The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.
spellingShingle Original Research
Li, Xuesong
Gao, Lei
Zheng, Longbin
Kou, Jiayuan
Zhu, Xing
Jiang, Yueqing
Zhong, Zhaoyu
Dan, Juhua
Xu, Haobo
Yang, Yang
Li, Hong
Shi, Sa
Cao, Wenwu
Zhao, Yajun
Tian, Ye
Yang, Liming
The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title_full The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title_fullStr The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title_full_unstemmed The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title_short The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages
title_sort efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in thp-1 macrophages
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309797/
https://www.ncbi.nlm.nih.gov/pubmed/25653524
http://dx.doi.org/10.2147/IJN.S75398
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