The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells
Abstract Background Graphene oxide (GO) nanoparticles (NPs) have been widely applied in various fields, especially in biomedical applications. Extensive studies have suggested that GO can pass through the blood–brain barrier (BBB) and induce abnormal autophagy and cytotoxicity in the central nervous...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-03-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12951-020-00605-6 |
| _version_ | 1798041449216868352 |
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| author | Chao Zhang Xiaoli Feng Longwen He Yaqing Zhang Longquan Shao |
| author_facet | Chao Zhang Xiaoli Feng Longwen He Yaqing Zhang Longquan Shao |
| author_sort | Chao Zhang |
| collection | DOAJ |
| description | Abstract Background Graphene oxide (GO) nanoparticles (NPs) have been widely applied in various fields, especially in biomedical applications. Extensive studies have suggested that GO can pass through the blood–brain barrier (BBB) and induce abnormal autophagy and cytotoxicity in the central nervous system (CNS). However, the effect and specific mechanism of GO on astrocytes, the most abundant cells in the brain still has not been extensively investigated. Results In this study, we systematically explored the toxicity and mechanism of GO exposure in the rat astroglioma-derived F98 cell line using molecular biological techniques (immunofluorescence staining, flow cytometry and Western blot) at the subcellular level and the signaling pathway level. Cells exposed to GO exhibited decreased cell viability and increased lactate dehydrogenase (LDH) release in a concentration- and time-dependent manner. GO-induced autophagy was evidenced by transmission electron microscopy (TEM) and immunofluorescence staining. Western blots showed that LC3II/I and p62 were upregulated and PI3K/Akt/mTOR was downregulated. Detection of lysosomal acidity and cathepsin B activity assay indicated the impairment of lysosomal function. Annexin V-FITC-PI detection showed the occurrence of apoptosis after GO exposure. The decrease in mitochondrial membrane potential (MMP) with an accompanying upregulation of cleaved caspase-3 and Bax/Bcl-2 further suggested that endogenous signaling pathways were involved in GO-induced apoptosis. Conclusion The exposure of F98 cells to GO can elicit concentration- and time-dependent toxicological effects. Additionally, increased autophagic response can be triggered after GO treatment and that the blocking of autophagy flux plays a vital role in GO cytotoxicity, which was determined to be related to dysfunction of lysosomal degradation. Importantly, the abnormal accumulation of autophagic substrate p62 protein can induce capase-3-mediated apoptosis. Inhibition of abnormal accumulation of autophagic cargo could alleviate the occurrence of GO-induced apoptosis in F98 cells. |
| first_indexed | 2024-04-11T22:21:41Z |
| format | Article |
| id | doaj.art-35b054e703944c178fb472ae0449cc0a |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-11T22:21:41Z |
| publishDate | 2020-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-35b054e703944c178fb472ae0449cc0a2022-12-22T04:00:05ZengBMCJournal of Nanobiotechnology1477-31552020-03-0118111710.1186/s12951-020-00605-6The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cellsChao Zhang0Xiaoli Feng1Longwen He2Yaqing Zhang3Longquan Shao4Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital)Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital)Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital)Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital)Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital)Abstract Background Graphene oxide (GO) nanoparticles (NPs) have been widely applied in various fields, especially in biomedical applications. Extensive studies have suggested that GO can pass through the blood–brain barrier (BBB) and induce abnormal autophagy and cytotoxicity in the central nervous system (CNS). However, the effect and specific mechanism of GO on astrocytes, the most abundant cells in the brain still has not been extensively investigated. Results In this study, we systematically explored the toxicity and mechanism of GO exposure in the rat astroglioma-derived F98 cell line using molecular biological techniques (immunofluorescence staining, flow cytometry and Western blot) at the subcellular level and the signaling pathway level. Cells exposed to GO exhibited decreased cell viability and increased lactate dehydrogenase (LDH) release in a concentration- and time-dependent manner. GO-induced autophagy was evidenced by transmission electron microscopy (TEM) and immunofluorescence staining. Western blots showed that LC3II/I and p62 were upregulated and PI3K/Akt/mTOR was downregulated. Detection of lysosomal acidity and cathepsin B activity assay indicated the impairment of lysosomal function. Annexin V-FITC-PI detection showed the occurrence of apoptosis after GO exposure. The decrease in mitochondrial membrane potential (MMP) with an accompanying upregulation of cleaved caspase-3 and Bax/Bcl-2 further suggested that endogenous signaling pathways were involved in GO-induced apoptosis. Conclusion The exposure of F98 cells to GO can elicit concentration- and time-dependent toxicological effects. Additionally, increased autophagic response can be triggered after GO treatment and that the blocking of autophagy flux plays a vital role in GO cytotoxicity, which was determined to be related to dysfunction of lysosomal degradation. Importantly, the abnormal accumulation of autophagic substrate p62 protein can induce capase-3-mediated apoptosis. Inhibition of abnormal accumulation of autophagic cargo could alleviate the occurrence of GO-induced apoptosis in F98 cells.http://link.springer.com/article/10.1186/s12951-020-00605-6Graphene oxideAstrocytep62AutophagyApoptosis |
| spellingShingle | Chao Zhang Xiaoli Feng Longwen He Yaqing Zhang Longquan Shao The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells Journal of Nanobiotechnology Graphene oxide Astrocyte p62 Autophagy Apoptosis |
| title | The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells |
| title_full | The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells |
| title_fullStr | The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells |
| title_full_unstemmed | The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells |
| title_short | The interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62-dependent apoptosis of F98 cells |
| title_sort | interrupted effect of autophagic flux and lysosomal function induced by graphene oxide in p62 dependent apoptosis of f98 cells |
| topic | Graphene oxide Astrocyte p62 Autophagy Apoptosis |
| url | http://link.springer.com/article/10.1186/s12951-020-00605-6 |
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