PMID- 35328525 OWN - NLM STAT- MEDLINE DCOM- 20220408 LR - 20220408 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 23 IP - 6 DP - 2022 Mar 13 TI - Activation of Autophagy by Low-Dose Silica Nanoparticles Enhances Testosterone Secretion in Leydig Cells. LID - 10.3390/ijms23063104 [doi] LID - 3104 AB - Silica nanoparticles (SNPs) can cause abnormal spermatogenesis in male reproductive toxicity. However, the toxicity and toxicological mechanisms of SNPs in testosterone synthesis and secretion in Leydig cells are not well known. Therefore, this study aimed to determine the effect and molecular mechanism of low doses of SNPs in testosterone production in Leydig cells. For this, mouse primary Leydig cells (PLCs) were exposed to 100 nm Stober nonporous spherical SNPs. We observed significant accumulation of SNPs in the cytoplasm of PLCs via transmission electron microscopy (TEM). CCK-8 and flow cytometry assays confirmed that low doses (50 and 100 mug/mL) of SNPs had no significant effect on cell viability and apoptosis, whereas high doses (more than 200 mug/mL) decreased cell viability and increased cell apoptosis in PLCs. Monodansylcadaverine (MDC) staining showed that SNPs caused the significant accumulation of autophagosomes in the cytoplasm of PLCs. SNPs activated autophagy by upregulating microtubule-associated protein light chain 3 (LC3-II) and BCL-2-interacting protein (BECLIN-1) levels, in addition to downregulating sequestosome 1 (SQSTM1/P62) level at low doses. In addition, low doses of SNPs enhanced testosterone secretion and increased steroidogenic acute regulatory protein (StAR) expression. SNPs combined with rapamycin (RAP), an autophagy activator, enhanced testosterone production and increased StAR expression, whereas SNPs combined with 3-methyladenine (3-MA) and chloroquine (CQ), autophagy inhibitors, had an opposite effect. Furthermore, BECLIN-1 depletion inhibited testosterone production and StAR expression. Altogether, our results demonstrate that low doses of SNPs enhanced testosterone secretion via the activation of autophagy in PLCs. FAU - Zhang, Jinlong AU - Zhang J AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. AD - Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China. FAU - Ye, Rongrong AU - Ye R AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. FAU - Grunberger, Jason William AU - Grunberger JW AUID- ORCID: 0000-0003-0436-5501 AD - Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112, USA. AD - Utah Center for Nanomedicine, University of Utah, Salt Lake City, UT 84112, USA. FAU - Jin, Jiaqi AU - Jin J AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. FAU - Zhang, Qianru AU - Zhang Q AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. FAU - Mohammadpour, Raziye AU - Mohammadpour R AD - Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112, USA. AD - Utah Center for Nanomedicine, University of Utah, Salt Lake City, UT 84112, USA. FAU - Khurana, Nitish AU - Khurana N AD - Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112, USA. AD - Utah Center for Nanomedicine, University of Utah, Salt Lake City, UT 84112, USA. FAU - Xu, Xianyu AU - Xu X AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. AD - Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China. FAU - Ghandehari, Hamidreza AU - Ghandehari H AD - Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112, USA. AD - Utah Center for Nanomedicine, University of Utah, Salt Lake City, UT 84112, USA. AD - Department of Biomedical Engineering, University of Utah, Salt Lake City, UT 84112, USA. FAU - Chen, Fenglei AU - Chen F AUID- ORCID: 0000-0002-7433-5474 AD - College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China. AD - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. AD - Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China. LA - eng GR - 31702298/National Natural Science Foundation of China/ GR - R01ES024681/the United States National Institute of Environmental Health Sciences/ GR - BK20170498/Natural Science Foundation of Jiangsu Province, China/ GR - 2017M621843/China Postdoctoral Science Foundation/ PT - Journal Article DEP - 20220313 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Beclin-1) RN - 0 (Microtubule-Associated Proteins) RN - 3XMK78S47O (Testosterone) RN - 7631-86-9 (Silicon Dioxide) SB - IM MH - Animals MH - Autophagy MH - Beclin-1/metabolism MH - *Leydig Cells/metabolism MH - Male MH - Mice MH - Microtubule-Associated Proteins/genetics/metabolism MH - *Nanoparticles MH - Silicon Dioxide/pharmacology MH - Testosterone/metabolism PMC - PMC8949068 OTO - NOTNLM OT - Leydig cells OT - StAR OT - autophagy OT - silica nanoparticles OT - testicular toxicity OT - testosterone COIS- The authors declare no conflict of interest. EDAT- 2022/03/26 06:00 MHDA- 2022/04/09 06:00 PMCR- 2022/03/13 CRDT- 2022/03/25 01:07 PHST- 2022/02/19 00:00 [received] PHST- 2022/03/10 00:00 [revised] PHST- 2022/03/11 00:00 [accepted] PHST- 2022/03/25 01:07 [entrez] PHST- 2022/03/26 06:00 [pubmed] PHST- 2022/04/09 06:00 [medline] PHST- 2022/03/13 00:00 [pmc-release] AID - ijms23063104 [pii] AID - ijms-23-03104 [pii] AID - 10.3390/ijms23063104 [doi] PST - epublish SO - Int J Mol Sci. 2022 Mar 13;23(6):3104. doi: 10.3390/ijms23063104.