PMID- 32547023 OWN - NLM STAT- MEDLINE DCOM- 20200730 LR - 20220415 IS - 1178-2013 (Electronic) IS - 1176-9114 (Print) IS - 1176-9114 (Linking) VI - 15 DP - 2020 TI - Silver Nanoparticle-Induced Apoptosis in ARPE-19 Cells Is Inhibited by Toxoplasma gondii Pre-Infection Through Suppression of NOX4-Dependent ROS Generation. PG - 3695-3716 LID - 10.2147/IJN.S244785 [doi] AB - PURPOSE: External and internal stimuli easily affect the retina. Studies have shown that cells infected with Toxoplasma gondii are resistant to multiple inducers of apoptosis. Nanoparticles (NPs) have been widely used in biomedical fields; however, little is known about cytotoxicity caused by NPs in the retina and the modulators that inhibit nanotoxicity. MATERIALS AND METHODS: ARPE-19 cells from human retinal pigment epithelium were treated with silver nanoparticles (AgNPs) alone or in combination with T. gondii. Then, the cellular toxicity, apoptosis, cell cycle analysis, autophagy, ROS generation, NOX4 expression, and MAPK/mTOR signaling pathways were investigated. To confirm the AgNP-induced cytotoxicity in ARPE-19 cells and its modulatory effects caused by T. gondii infection, the major experiments carried out in ARPE-19 cells were performed again using human foreskin fibroblast (HFF) cells and bone marrow-derived macrophages (BMDMs) from NOX4(-/) (-) mice. RESULTS: AgNPs dose-dependently induced cytotoxicity and cell death in ARPE-19 cells. Apoptosis, sub-G1 phase cell accumulation, autophagy, JNK phosphorylation, and mitochondrial apoptotic features, such as caspase-3 and PARP cleavages, mitochondrial membrane potential depolarization, and cytochrome c release into the cytosol were observed in AgNP-treated cells. AgNP treatment also increased the Bax, Bik, and Bim protein levels as well as NOX4-dependent ROS generation. However, T. gondii-infected ARPE-19 cells inhibited AgNP-induced apoptosis, JNK phosphorylation, sub-G1 phase cell accumulation, autophagy, NOX4-mediated ROS production, and mitochondrial apoptosis. Furthermore, mitochondrial apoptosis was found in AgNP-treated HFF cells and BMDMs, and AgNP-induced mitochondrial apoptosis inhibition via NOX4-dependent ROS suppression in T. gondii pre-infected HFF cells and BMDMs was also confirmed. CONCLUSION: AgNPs induced mitochondrial apoptosis in human RPE cells combined with cell cycle dysregulation and autophagy; however, these effects were significantly inhibited by T. gondii pre-infection by suppression of NOX4-mediated ROS production, suggesting that T. gondii is a strong inhibitory modulator of nanotoxicity in in vitro models. CI - (c) 2020 Quan et al. FAU - Quan, Juan-Hua AU - Quan JH AD - Department of Gastroenterology, The Affiliated Hospital of Guangdong Medical University, Zhanjiang 524-001, People's Republic of China. FAU - Gao, Fei Fei AU - Gao FF AUID- ORCID: 0000-0002-4454-8073 AD - Department of Infection Biology and Department of Medical Science, Chungnam National University College of Medicine, Daejeon 301-131, Korea. FAU - Ismail, Hassan Ahmed Hassan Ahmed AU - Ismail HAHA AD - Communicable and Non-Communicable Diseases Control Directorate, Federal Ministry of Health, Khartoum, Sudan. FAU - Yuk, Jae-Min AU - Yuk JM AUID- ORCID: 0000-0002-0630-9194 AD - Department of Infection Biology and Department of Medical Science, Chungnam National University College of Medicine, Daejeon 301-131, Korea. FAU - Cha, Guang-Ho AU - Cha GH AD - Department of Infection Biology and Department of Medical Science, Chungnam National University College of Medicine, Daejeon 301-131, Korea. FAU - Chu, Jia-Qi AU - Chu JQ AD - Stem Cell Research and Cellular Therapy Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong Province 524-001, People's Republic of China. FAU - Lee, Young-Ha AU - Lee YH AUID- ORCID: 0000-0002-5795-8960 AD - Department of Infection Biology and Department of Medical Science, Chungnam National University College of Medicine, Daejeon 301-131, Korea. LA - eng PT - Journal Article DEP - 20200526 PL - New Zealand TA - Int J Nanomedicine JT - International journal of nanomedicine JID - 101263847 RN - 0 (Reactive Oxygen Species) RN - 3M4G523W1G (Silver) RN - EC 1.6.3.- (NADPH Oxidase 4) RN - EC 1.6.3.- (NOX4 protein, human) SB - IM MH - Animals MH - Apoptosis/*drug effects MH - Autophagy/drug effects MH - Cell Line MH - Cell Shape/drug effects MH - Disease Models, Animal MH - Fibroblasts/drug effects/parasitology MH - G1 Phase/drug effects MH - Humans MH - MAP Kinase Signaling System/drug effects MH - Male MH - Metal Nanoparticles/*chemistry MH - Mice MH - Mitochondria/drug effects/metabolism MH - Models, Biological MH - NADPH Oxidase 4/*metabolism MH - Phosphorylation/drug effects MH - Reactive Oxygen Species/*metabolism MH - Retinal Pigment Epithelium/*metabolism/*parasitology MH - Silver/*pharmacology MH - Toxoplasmosis/*pathology PMC - PMC7266428 OTO - NOTNLM OT - NADPH oxidase 4 OT - Toxoplasma gondii OT - human retinal pigment epithelium OT - mitochondrial apoptosis OT - reactive oxygen species OT - silver nanoparticles COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2020/06/18 06:00 MHDA- 2020/07/31 06:00 PMCR- 2020/05/26 CRDT- 2020/06/18 06:00 PHST- 2020/06/18 06:00 [entrez] PHST- 2020/06/18 06:00 [pubmed] PHST- 2020/07/31 06:00 [medline] PHST- 2020/05/26 00:00 [pmc-release] AID - 244785 [pii] AID - 10.2147/IJN.S244785 [doi] PST - epublish SO - Int J Nanomedicine. 2020 May 26;15:3695-3716. doi: 10.2147/IJN.S244785. eCollection 2020.