Evaluation of Insulin-Like Activity of Novel Zinc Metal–Organics toward Adipogenesis Signaling
Abstract
:1. Introduction
2. Results
2.1. Synthesis
2.2. Description of Crystallographic Structures
2.3. ESI-MS Spectrometry
2.4. Solution NMR Spectroscopy
2.4.1. 13C-NMR Spectroscopy
2.4.2. 1H-NMR Solution NMR
2.5. Thermal Studies
2.6. Luminescence
2.7. Biological Studies
2.7.1. Cytotoxicity Results
Zn(II) Compound Toxicity in 3T3-L1 Pre-Adipocytes for 24 h
Zn(II) Compound Toxicity in 3T3-L1 Pre-Adipocytes for 48 h
Zn(II) Compound Toxicity in Differentiated 3T3-L1 Cells for 48 h
2.7.2. Oil Red O Staining Results
2.7.3. Biomarker Gene Expression
2.8. Cell Signaling Studies
3. Discussion
3.1. From Hydroxycarboxylic Acids to Zwitterions in Zinc Chemistry
3.2. Structural Specificity of Zinc in the Biology of Adipogenesis
3.3. Luminescence
3.4. The Insulin Mimetic Potential of Hybrid Zinc Metal–Organics
4. Materials and Methods
4.1. Physical Measurements
4.2. Syntheses
4.2.1. Synthesis of Zn(C7H7NO2)2Cl2 (1)
4.2.2. Synthesis of [Zn(C6H9NO5)(H2O)]n • nH2O (2)
4.2.3. Synthesis of [Zn(C6H9NO5)(C12H8N2)]•4H2O (3)
4.2.4. Synthesis of Zn(C12H8N2)Cl2 (4)
4.2.5. Synthesis of [Zn(C12H8N2)2Cl](NO3)•H2O (5)
4.3. X-ray Crystal Structure Determination
4.4. Cell Cultures and Biological Tests
4.4.1. Cultivation of 3T3-L1 Cells
4.4.2. Preparation of Zn(II) Compound Stock Solutions
4.4.3. Cell Viability Growth Studies
4.4.4. Cell Biocompatibility—Morphology—Migration
4.4.5. Induction of Adipogenesis
4.4.6. Zn(II)-Induced Adipogenesis
4.4.7. Oil Red O Staining
4.4.8. cDNA Synthesis and RT-PCR Assay
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
Abbreviation List
ANOVA | One-way analysis of variance |
DM | Diabetes mellitus |
DMEM | Dulbecco’s modified Eagle’s medium |
FBS | Fetal Bovine Serum |
GLUT 1,4 | Glucose transporter 1,4 |
PPAR-γ | Peroxisome proliferator-activated receptor gamma |
SEM | Standard error mean |
LMCT | Ligand-to-metal-charge-transfer |
ADA | American Diabetes Association |
ESI-MS | Electrospray ionization mass spectrometry |
TGA | Thermogravimetric analysis |
RLU | Relative luminescence units |
MAPK | Mitogen-activated protein kinase |
PI3K | Phosphoinositide 3-kinase |
NMR | Nuclear magnetic resonance |
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1 | 2 | 3 | 4 | 5 | |
---|---|---|---|---|---|
Formula | C14H14Cl2N2O4Zn | C6H13NO7Zn | C18H25N3O9Zn | C12H8Cl2N2Zn | C24H18ClN5O4Zn |
Fw | 410.54 | 276.54 | 492.78 | 316.47 | 541.25 |
Space group | P | P21/c | P | P21/n | P21/n |
a (Å) | 7.2582(1) | 7.3509(3) | 7.0736(1) | 7.9667(1) | 10.8589(2) |
b (Å) | 8.1539(1) | 9.7341(2) | 12.4910(2) | 15.6568(2) | 15.4397(2) |
c (Å) | 13.8300(3) | 14.3156(3) | 12.7785(2) | 9.6838(1) | 13.1342(2) |
α(o) | 83.107(1) | 90.0 | 67.493(1) | 90.0 | 90.0 |
β(o) | 84.445(1) | 99.807(1) | 82.002(1) | 102.044(1) | 93.997(1) |
γ(o) | 75.707(1) | 90.0 | 79.475(1) | 90.0 | 90.0 |
V (Å3) | 785.51(2) | 1009.38(5) | 1022.53(3) | 1181.30(2) | 2196.70(6) |
Z | 2 | 4 | 2 | 4 | 4 |
T (°C) | −113 | −113 | −113 | −113 | −113 |
Radiation | Cu Kα | Cu Kα | Cu Kα | Cu Kα | Cu Kα |
ρcalcd (g cm−3) | 1.736 | 1.820 | 1.600 | 1.779 | 1.637 |
μ (mm−1) | 5.500 | 3.622 | 2.187 | 6.840 | 3.056 |
Reflections with I 2σ(I) | 2016 | 1347 | 2991 | 1848 | 3535 |
Ra | 0.0362 | 0.0556 | 0.0316 | 0.0271 | 0.0350 |
Rwa | 0.0787 | 0.1085 | 0.0781 | 0.0703 | 0.0909 |
Distances (Å) | |||||||||
---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | |||||
Zn-O(1) | 1.966(2) | Zn-O(4) | 2.016(3) | Zn(1)-N(12) | 2.086(2) | Zn(1)-N(2) | 2.059(2) | Zn-N(21) | 2.068(2) |
Zn-O(11) | 1.967(2) | Zn-O(1) | 2.074(3) | Zn(1)-O(3) | 2.088(2) | Zn(1)-N(1) | 2.078(2) | Zn-N(1) | 2.076(2) |
Zn-Cl(2) | 2.250(1) | Zn-O(1w) | 2.108(4) | Zn(1)-O(1) | 2.088(2) | Zn(1)-Cl(2) | 2.2102(6) | Zn-N(2) | 2.148(2) |
Zn-Cl(1) | 2.269(1) | Zn-O(3) | 2.134(3) | Zn(1)-N(1) | 2.144(2) | Zn(1)-Cl(1) | 2.2190(6) | Zn-N(22) | 2.179(2) |
Zn-O(5) | 2.145(3) | Zn(1)-N(11) | 2.199(2) | Zn-Cl | 2.2951(6) | ||||
Zn-N | 2.201(3) | Zn(1)-O(5) | 2.244(2) | ||||||
Angles (o) | |||||||||
O(1)-Zn-O(11) | 113.52(10) | O(4)-Zn-O(1) | 110.84(11) | N(12)-Zn(1)-O(3) | 90.01(7) | N(2)-Zn(1)-N(1) | 81.49(8) | N(21)-Zn-N(1) | 121.09(7) |
O(1)-Zn-Cl(2) | 105.80(8) | O(4)-Zn-O(1w) | 92.99(12) | N(12)-Zn(1)-O(1) | 107.62(7) | N(2)-Zn(1)-Cl(2) | 108.31(6) | N(21)-Zn-N(2) | 96.90(7) |
O(11)-Zn-Cl(2) | 112.04(9) | O(1)-Zn-O(1w) | 88.78(13) | O(3)-Zn(1)-O(1) | 101.41(6) | N(1)-Zn(1)-Cl(2) | 125.83(6) | N(1)-Zn-N(2) | 78.94(7) |
O(1)-Zn-Cl(1) | 111.22(9) | O(4)-Zn-O(3) | 93.25(11) | N(12)-Zn(1)-N(1) | 171.07(8) | N(2)-Zn(1)-Cl(1) | 115.77(5) | N(21)-Zn-N(22) | 78.72(7) |
O(11)-Zn-Cl(1) | 104.54(8) | O(1)-Zn-O(3) | 155.57(11) | O(3)-Zn(1)-N(1) | 82.41(6) | N(1)-Zn(1)-Cl(1) | 107.11(5) | N(1)-Zn-N(22) | 98.24(7) |
Cl(2)-Zn-Cl(1) | 109.80(4) | O(1w)-Zn-O(3) | 85.85(14) | O(1)-Zn(1)-N(1) | 78.66(7) | Cl(2)-Zn(1)-Cl(1) | 114.40(2) | N(2)-Zn-N(22) | 172.72(7) |
O(4)-Zn-O(5) | 88.91(12) | N(12)-Zn(1)-N(11) | 78.29(7) | N(21)-Zn-Cl | 118.31(5) | ||||
O(1)-Zn-O(5) | 96.16(12) | O(3)-Zn(1)-N(11) | 166.26(7) | N(1)-Zn-Cl | 120.59(5) | ||||
O(1w)-Zn-O(5) | 173.70(14) | N(1)-Zn(1)-N(11) | 108.58(7) | N(2)-Zn-Cl | 95.16(5) | ||||
O(3)-Zn-O(5) | 88.06(12) | N(12)-Zn(1)-O(5) | 95.65(7) | N(22)-Zn-Cl | 92.04(5) | ||||
O(4)-Zn-N | 166.17(12) | O(3)-Zn(1)-O(5) | 87.80(6) | ||||||
O(1)-Zn-N | 78.18(11) | O(1)-Zn(1)-O(5) | 154.81(7) | ||||||
O(1w)-Zn-N | 97.77(13) | N(1)-Zn(1)-O(5) | 79.42(7) | ||||||
O(3)-Zn-N | 78.96(11) | N(11)-Zn(1)-O(5) | 86.24(6) | ||||||
O(5)-Zn-N | 79.49(12) |
Gene | Primer (Forward (F); Reverse (R)) | Sequence |
---|---|---|
GLUT 4 | F | 5′-AACCAGCATCTTCGAGTCGG-3′ |
R | 5′-TAAGAGCACCGAGACCAACG-3′ | |
GLUT 1 | F | 5′-CTCACCACGCTTTGGTCTCT-3′ |
R | 5′-CGCCTGCCAAAGCGATTAAC-3′ | |
PPAR-γ | F | 5′-GTCAGCGACTGGGACTTTTC-3′ |
R | 5′-CGAGGACATCCAAGACAACC-3′ |
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Gabriel, C.; Tsave, O.; Yavropoulou, M.P.; Architektonidis, T.; Raptopoulou, C.P.; Psycharis, V.; Salifoglou, A. Evaluation of Insulin-Like Activity of Novel Zinc Metal–Organics toward Adipogenesis Signaling. Int. J. Mol. Sci. 2021, 22, 6757. https://doi.org/10.3390/ijms22136757
Gabriel C, Tsave O, Yavropoulou MP, Architektonidis T, Raptopoulou CP, Psycharis V, Salifoglou A. Evaluation of Insulin-Like Activity of Novel Zinc Metal–Organics toward Adipogenesis Signaling. International Journal of Molecular Sciences. 2021; 22(13):6757. https://doi.org/10.3390/ijms22136757
Chicago/Turabian StyleGabriel, Catherine, Olga Tsave, Maria P. Yavropoulou, Theodore Architektonidis, Catherine P. Raptopoulou, Vassilis Psycharis, and Athanasios Salifoglou. 2021. "Evaluation of Insulin-Like Activity of Novel Zinc Metal–Organics toward Adipogenesis Signaling" International Journal of Molecular Sciences 22, no. 13: 6757. https://doi.org/10.3390/ijms22136757