Table 1.
Major in vitro studies portraying associations between BPA and phthalates, and obesity.
| Authors, Year | Type of Cell Culture | Main Findings | Remarks |
|---|---|---|---|
| Bisphenol A and obesity | |||
| Riu et al., 2011 [152] | NIH3T3-L1 cell line (pre-adipocytes) |
1. ↑ adipogenesis 2. ↑ lipid accumulation 3. ↑ mRNA level of PPARγ 4. ↑ PPARγ activity |
1. Animal in vitro model 2. ED: TBBPA 3. Obesogenic effects at 10 µM |
| Valentino et al., 2013 [118] | Primary hADSCs | 1. (-) mRNA level of PPARγ, GLUT4 2. ↓ of glucose utilization 3. ↓ tyrosine phosphorylation of insulin receptor (IR) 3. ↓ of PKB/Akt phosphorylation 4. ↑ of IL-6, IFN-γ 5. ↑ of JAK/STAT, JNK 6. ↑ activity NF-kB pathway |
1. Human in vitro model 2. ED: BPA 3. Biological effects at 1 nM |
| Bastos Sales et al., 2013 [102] | Murine N2A, human SK-N-AS neuroblastoma cells and murine pre-adipocyte fibroblasts (3T3-L1) | 1. Modest ↓ in global DNA methylation in murine N2A cells 2. No changes in global DNA methylation in human SK-N-AS cells. 3. ↑ adipocyte differentiation in murine 3T3-L1 pre-adipocytes |
1. Animal and human in vitro model 2. ED: BPA and a range of several EDCs not belonging to bisphenols 3. Biological effects at ≥ 10 μΜ |
| Menale et al., 2015 [137] | Primary pre-adipocytes | 1. ↑ adipogenesis 2. ↑ lipid accumulation 3. ↑ mRNA level of ERα (10 nM, 100 nM) 4. (-) mRNA level of ERβ 5. ↑ production of IL1B, IL18, CCL20 (10 nM) |
1. Human in vitro model 2. ED: BPA 3. Obesogenic effects at 1 nM, 10 nM, 100 nM |
| Ariemma et al., 2016 [92] | 3T3-L1 Pre-adipocytes |
1. Undifferentiated cells: - ↑ proliferation - ↑ differentiation - ↑ expression of PPARγ, C/EBPα and FABP4/AP2 2. Mature adipocytes: - Hypertrophy - ↑ lipid accumulation - ↑ mRNA of leptin, IL6, IFNγ - ↓ glucose utilization |
1. Animal in vitro model 2. ED: BPA 3. Obesogenic effects at 1 nM |
| Longo et al., 2020 [103] | 3T3L1 and NIH3T3 (committed and uncommitted pre-adipocytes, respectively) | - ↓ DNA methylation at PPARγ promoter, without affecting mRNA expression in pre-adipocytes - Transient ↑ in PPARγ expression and lipid accumulation at D4 of differentiation in 3T3L1 cells - Ending BPA exposure restores the PPARγ promoter methylation and inflammatory profile of 3T3L1 cells. - Expression of PPARγ is barely detectable and its promoter is completely methylated in NIH3T3 cells - ↑ PPARγ expression is more evident both in pre-adipocytes and during the adipocyte differentiation |
1. Animal in vitro model 2. ED: BPA 3. Biological effects at low doses: 1 nM |
| Cohen et al., 2021 [153] | Primary hADSCs | 1. ↑ adipogenesis and lipid production at 0.1 nM 2. ↓ adipogenesis and lipid production at 10 nM |
1. Human in vitro model 2. ED: BPA 3. Biological effects at 0.1 nM, 10 nM |
| Yamasaki et al., 2021 [154] | ST-13 cell line (pre-adipocytes) | - Undifferentiated cells: 1. (-) lipid accumulation 2. (-) mRNA level of PPARγ 3. ↑ mRNA level of AACS, PLIN1, FAS, CIDEA, LSD-1 - Mature adipocytes: 1. (-) lipid accumulation 2. (-) mRNA level of AACS, SCOT |
1. Animal in vitro model 2. ED: TBBPA 3. Obesogenic effects at 0.5 µM, 1 µM |
| Schaffert et al., 2021 [121] | SGBSs (pre-adipocytes) | 1. ↑ binding to PPARγ (50 µM) 2. (-) PPARγ activity (10 nM, 100 nM, 1 µM, 10 µM) 3. ↓ lipid accumulation (10 nM, 100 nM, 1 µM, 10 µM) 4. ↑ leptin (10 nM) 5. ↓ cellular ROS level (10 nM, 100 nM, 1 µM, 10 µM) 6. ↓ insulin sensitivity (1 µM) |
1. Human in vitro model 2. ED: BPA 3. Obesogenic effects at 10 nM, 100 nM, 1 µM, 10 µM, 50 µM |
| Marqueno et al., 2021 [155] | ZFL cell line (primary mouse hepatocytes) | 1. ↑ lipid accumulation (5 µM, 50 µM) 2. ↑ ROS generation (20 µM, 50 µM, 70 µM, 100 µM, 150 µM, 200 µM) |
1. Animal in vitro model 2. ED: BPA 3. Biological effects at 5 µM, 20 µM, 50 µM, 70 µM, 100 µM, 150 µM, 200 µM |
| Lee et al., 2022 [156] | Huh-7 cell line (primary hepatocytes) | 1. ↓ cell viability (200 µM, 400 µM) 2. ↑ lipid accumulation (10 µM, 50 µM, 100 µM, 200 µM) 3. Fatty acid uptake ↑ (10 µM, 50 µM, 100 µM) 4. ↑ intracellular ROS formation (10 µM, 50 µM, 100 µM, 200 µM) |
1. Human in vitro model 2. ED: BPA 3. Biological effects at 10 µM, 50 µM, 100 µM, 200 µM, 400 µM |
| Phthalates and obesity | |||
| Sargis et al., 2010 [85] | 3T3-L1 cell line (pre-adipocytes) | 1. ↑ adipogenic differentiation 2. ↑ lipid accumulation (100 nM) 3. ↑ PPARγ and glucocorticoid-like activity (1 µM) 4. ↑ adiponectin and protein expression of IR-β (1 µM–100 pM) |
1. Animal in vitro model 2. Pthalate: DCHP 3. Obesogenic effects at 100 pM, 1 nM, 10 nM, 100 nM, 1 µM |
| Dimastrogiovanni et al., 2015 [150] | RTL-W1 cell line (hepatocytes) |
1. ↑ lipid accumulation 2. ↓ alteration of membrane lipids 3. ↓ mRNA level of CD36, FAS, LPL |
1. Animal in vitro model 2. Pthalate: DEHP 3. Biological effects at 5 μΜ |
| Zhang et al., 2017 [105] | C3H10T1/2 cell line (MSCs) | 1. ↑ adipogenesis 2. ↑ mRNA level of AP2, PPARγ 3. ↑ lipid accumulation 4. ↑ protein level of FOXO1 5. ↑ acetylation of FOXO1, β-catenin 6. ↓ protein level of SIRT1, SIRT3 |
1. Animal in vitro model 2. Pthalate: BBP 3. Biological effects at 50 μΜ |
| Schaedlich et al., 2018 [151] | SGBSs (pre-adipocytes) | 1. ↓ TGsaccumulation 2. ↓ adiponectin production 3. ↓ protein level of PPARα, PPARγ 4. ↓ phosphorylation of ERK1, ERK2 5. ↑ lipolysis 6. ↑ ROS formation |
1. Human in vitro model 2. Pthalate: DEHP 3. Obesogenic effects at 50 µg/mL |
| Zhang et al., 2019 [145] | BRL-3A cell line (hepatocytes) |
1. ↑ lipid accumulation (100 µM, 200 µM) 2. ↑ mRNA level of FAS, PDK4, AP2 (10 µM, 50 µM, 100 µM, 200 µM) 3. ↑ mRNA level of PPARγ (50 µM, 100 µM, 200 µM) 4. ↓ JAK2/STAT5 signaling 5. ↓ level of indicators of oxidative stress: SOD ↓, MDA ↑ (10 µM, 50 µM, 100 µM, 200 µM) |
1. Animal in vitro model 2. Pthalate: MEHP 3. Biological effects at 10 µM, 50 µM, 100 µM, 200 µM |
| Perez-Albaladejo et al., 2021 [157] | PLHC-1 cell line (hepatocytes) | - DBP: 1. ↑ TG accumulation (20 µM) 2. ↑ ROS formation (5 µM, 20 µM, 50 µM, 100 µM) - DEHP: 1. ↑ TG accumulation (5 µM, 10 µM) 2. ↑ ROS formation (100 µM) |
1. Animal in vitro model 2. Phthalates: DBP and DEHP 3. Biological effects at - DBP: 5 µM, 20 µM, 50 µM, 100 µM - DEHP: 5 µM, 10 µM, 100 µM |
| Meruvu et al., 2021 [104] | 3T3-L1 cells | - ↑ miR-34a-5p expression - ↑ adipogenesis - ↓ Nampt, Sirt1 and Sirt3 gene expression levels; ↓ Nampt protein - ↓ adipogenesis, ↑ Nampt protein and NAD+ after miR-34a-5p knockdown in the presence of BBP |
1. Animal in vitro model 2. Phthalate: BBP 2. Biological effects at various doses of BBP without exogenous adipogenic stimuli |
| Al-Abdulla et al., 2022 [158] | MIN-6 cell line (pancreatic cells) | 1. ↓ viability of cells after 24 exposure at 1 μΜ 2. ↑ mRNA level of SUR1, GLUT2 at 10 μΜ 3. ↓ GSIS (20 μΜ glucose) 4. ↓ insulin content at 1 μΜ |
1. Animal in vitro model 2. Pthalate: DEHP 3. Dose: 100 pM, 1 nM, 10 nM, 100 nM, 1 µM, 10 µM |
| Schaffert et al., 2022 [141] | SGBSs (pre-adipocytes) |
1. DINP: - ↑ binding to PPARγ - (-) PPARγ activation - (-) lipid accumulation - ↑ adipsin (10 µM) - Mature adipocytes: * 10 µM: ↑ MCP-1, LAP3, GPX1 * 10 nM: ↑ GPX8, GSR * 10 nM, 10 µM: ↑ LEP, GPX4 * 10 nM, 10 µM: ↓ adiponectin 2. DPHP: - ↑ binding to PPARγ - (-) PPARγ activation - Undifferentiated cells: * (-) lipid accumulation * ↓ MCP-1 (10 nM, 10 µM) - Mature adipocytes: * ↓ lipid accumulation (10 µM, 25 µM, 50 µM, 100 µM) * 10 µM: ↑ LEP, MCP-1, LAP-3, GPX4, GPX8, adipsin * 10 nM: ↑ GSR * 10 nM, 10 µM: ↑ GPX1, GSTO1 * 10 nM, 10 µM: ↓ adiponectin 3. MHINP: - ↑ binding to PPARγ (100 µM, 200 µM, 400 µM) - ↑ PPARγ activation (1 µM) - Undifferentiated cells: * ↑ pre-adipocyte differentiation, lipid accumulation (10 µM, 25 µM, 50 µM, 100 µM) * 10 µM: ↑ LEP, PLIN1, GPD1, FASN, FABP4, FABP5 * 10 nM: ↓ MCP-1 * 10 nM, 10 µM: ↑ adipsin - Mature adipocytes: * 1 µM: ↑ lipid accumulation * 10 µM: ↑ LAP3, adipsin * 10 nM: ↑ GSR, GPX8 * 10 nM, 10 µM: ↑LEP, MCP-1, GPX1, GPX4, GSTO1 * 10 nM, 10 µM: ↓ adiponectin 4. OH-MPHP: - ↑ binding to PPARγ - ↑ PPARγ activation - Undifferentiated cells: * ↑ pre-adipocyte differentiation, lipid accumulation (10 µM, 25 µM, 50 µM) * ↑ LEP, GPD1, FASN, FABP4, FABP5 (10 µM) - Mature adipocytes: * 10 µM: ↑ LAP3, GPX1, GPX4, GPX8, adipsin * 10 nM, 10 µM: ↑ LEP, GSR, MCP-1, GSTO1 * 10 nM, 10 µM: ↓ adiponectin * 10 nM, 10 µM, 25 µM, 50 µM, 100 µM: ↓ lipid accumulation |
1. Human in vitro model 2. Phthalates: DINP, DPHP, MHINP, OH-MPHP 2. Obesogenic effects at - DINP: 10 nM, 10 µM, - DPHP: 10 nM, 10 µM, 25 µM, 50 µM, 100 µM - MHINP: 10 nM, 10 µM, 25 µM, 50 µM, 100 µM, 200 µM, 400 µM - OH-MPHP: 10 nM, 10 µM, 25 µM, 50 µM, 100 µM, 200 µM, 400 µM |
Abbreviations: AACS: acetoacetyl-CoA synthetase; AP2: adipocyte protein 2; BPA: bisphenol A; BBP: benzyl butyl phthalate; CCL20: chemokine (C-C motif) ligand 20; C/EBPα: CCAAT/enhancer-binding protein α; CD36: fatty acid translocase; CIDEA: cell-death-inducing DNA fragmentation factor-alpha-like effector A; DCHP: bis(2-propylheptyl) phthalate; DBP: dibutyl phthalate; DEHP: di(2-ethylhexyl) phthalate; DINP: diisononyl phthalate; DPHP: bis(2-propylheptyl) phthalate; ED: endocrine disruptor; ERα: estrogen receptor α; ERβ: estrogen receptor β; ERK1/2: extracellular-signal-regulated protein kinase 1/2; FABP4: fatty-acid-binding protein 4; FABP5: fatty-acid-binding protein 5; FABP4/AP2: fatty-acid-binding protein 4/adipocyte protein 2; FASN: fatty acid synthase; FOXO1: forkhead box protein O1; GLUT2: glucose transporter type 2; GLUT4: glucose transporter type 4; GPD1: glycerol-3-phosphate-dehydrogenase; GPX1: glutathione peroxidase 1; GPX4: glutathione peroxidase 4; GPX8: glutathione peroxidase 8; GSIS: glucose-stimulated insulin secretion; GSR: glutathione-disulfide reductase; GSTO1: glutathione S-transferase omega-1; IFN-γ: interferon-γ; IL: interleukin; IR-β: insulin receptor subunit β; JAK/STAT: Janus kinase/signal transducer and activator of transcription; JNK: c-Jun N-terminal kinase;LAP3: leucine aminopeptidase 3; LEP: leptin; LPL: lipoprotein lipase; LSD-1: lysine-specific demethylase-1; MCP-1: monocyte chemoattractant protein-1; MDA: malondialdehyde; MEHP: mono-2-ethylhexyl phthalate; MHINP: monohydroxy isononyl phthalate; N2A: Neuro-2A cells; NAD: nicotinamide adenine dinucleotide; Nampt: nicotinamide phosphoribosyltransferase; NF-kB: nuclear factor kappa-light-chain-enhancer of activated B cells; OH-MPHP: 6-hydroxy monopropylheptyl phthalate; PDK4: pyruvate dehydrogenase kinase 4; PPARα: peroxisome-proliferator-activated receptor α; PPARγ: peroxisome-proliferator-activated receptor γ; PKB/Akt: protein kinase B/AKT; PLIN1: perilipin-1; ROS: reactive oxygen species; SCOT: succinyl-CoA-3-oxoacid CoA-transferase; SIRT1: sirtuin 1; SIRT3: sirtuin 3; SOD: superoxide dismutase; SUR1: sulfonylurea receptor 1; TBBPA: tetrabromobisphenol A; TGs: triglycerides. ↑ increase, ↓ decrease.