The Cooperative Effect of Genistein and Protein Hydrolysates on the Proliferation and Survival of Osteoblastic Cells (hFOB 1.19)
Abstract
:1. Introduction
2. Results
2.1. Degrees of Hydrolysis and Amino Acid Compositions of the Prepared Hydrolysates
2.2. Growth Proliferation and Inhibition of Genistein on the Osteoblasts
2.3. Cooperation between Genistein and the Hydrolysates in Osteoblast Proliferation
2.4. Cooperation between Genistein and the Hydrolysates in Osteoblast Anti-Apoptosis
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Preparation of De-Isoflavoned Soy Protein and Protein Hydrolysates
4.3. Cell Line and Culture Conditions
4.4. In Vitro Effect of Genistein on the Osteoblasts
4.5. Assay of Cell Proliferation
4.6. Apoptosis Assay
4.7. Chemical Analyses
4.8. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
Gen | genistein |
CH | casein hydrolysate |
GH | gelatin hydrolysate |
SH | soy protein hydrolysate |
DH | degree of hydrolysis |
CCK-8 | cell counting kit-8 |
DMSO | dimethyl sulfoxide |
E | 17β-estradiol |
WST-8 | 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt |
EDTA | ethylenediamine tetra-acetic acid |
EP | etoposide |
FACS | fluorescence-activated cell sorting |
FBS | fetal bovine serum |
hFOB 1.19 cells | human fetal osteoblastic cells |
OPA | o-phthaldialdehyde |
PBS | phosphate-buffered saline |
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Amino Acids | Hydrolysates | Amino Acids | Hydrolysates | ||||
---|---|---|---|---|---|---|---|
GH2 | SH2 | CH2 | GH2 | SH2 | CH2 | ||
Ala | 100.1 | 39.0 | 31.2 | Lys | 37.7 | 73.7 | 80.2 |
Arg | 90.0 | 87.6 | 34.8 | Met | 8.1 | 14.1 | 32.0 |
Asp | 54.1 | 128.1 | 77.2 | Phe | 22.6 | 52.1 | 51.2 |
Cys | ND | 14.2 | ND | Pro | 146.4 | 61.2 | 109.2 |
Glu | 112.9 | 245.5 | 229.5 | Ser | 34.7 | 59.8 | 59.8 |
Gly | 266.1 | 47.1 | 21.8 | Thr | 19.2 | 39.4 | 46.9 |
His | 6.2 | 25.5 | 27.3 | Trp | ND | 12.5 | 14.6 |
4-Hyp | 110.7 | ND | ND | Tyr | 0.5 | 34.8 | 56.7 |
Ile | 16.2 | 46.8 | 55.1 | Val | 31.5 | 44.5 | 67.3 |
Groups | Dose Levels | Treatment Times | |
---|---|---|---|
48 h | 72 h | ||
Gen | 2.5 μg/L | 109.4 ± 4.1 | 112.3 ± 6.0 |
17β-Estradiol | 10−8 mol/L | 134.2 ± 2.9 | 126.3 ± 3.5 |
GH1 | 0.02 g/L | 108.4 ± 2.6 | 109.4 ± 1.6 |
GH1 | 0.05 g/L | 121.8 ± 2.5 | 117.9 ± 2.2 |
GH1 | 0.1 g/L | 114.6 ± 1.9 | 113.2 ± 1.4 |
Gen-GH1 | 2.5 μg/L+ 0.02 g/L | 113.8 ± 1.9 | 115.1 ± 2.2 |
Gen-GH1 | 2.5 μg/L + 0.05 g/L | 127.2 ± 2.6 | 123.1 ± 2.8 |
Gen-GH1 | 2.5 μg/L + 0.1 g/L | 119.2 ± 1.3 | 117.9 ± 2.9 |
GH2 | 0.02 g/L | 122.7 ± 2.8 | 110.0 ± 1.4 |
GH2 | 0.05 g/L | 131.1 ± 4.3 | 125.3 ± 2.1 |
GH2 | 0.1 g/L | 124.0 ± 2.0 | 116.7 ± 1.4 |
Gen-GH2 | 2.5 μg/L + 0.02 g/L | 125.3 ± 3.4 | 115.8 ± 2.1 |
Gen-GH2 | 2.5 μg/L + 0.05 g/L | 140.9 ± 1.3 | 131.2 ± 4.1 |
Gen-GH2 | 2.5 μg/L + 0.1 g/L | 130.2 ± 2.3 | 122.2 ± 3.6 |
SH1 | 0.02 g/L | 113.2 ± 5.7 | 115.8 ± 6.2 |
SH1 | 0.05 g/L | 110.4 ± 4.1 | 110.9 ± 2.6 |
SH1 | 0.1 g/L | 104.6 ± 6.0 | 104.6 ± 0.8 |
Gen-SH1 | 2.5 μg/L + 0.02 g/L | 114.6 ± 4.5 | 118.7 ± 4.4 |
Gen-SH1 | 2.5 μg/L + 0.05 g/L | 111.1 ± 4.4 | 114.8 ± 4.7 |
Gen-SH1 | 2.5 μg/L + 0.1 g/L | 104.8 ± 3.2 | 108.7 ± 2.2 |
SH2 | 0.02 g/L | 119.8 ± 5.7 | 107.7 ± 3.3 |
SH2 | 0.05 g/L | 121.3 ± 2.4 | 110.6 ± 5.6 |
SH2 | 0.1 g/L | 118.0 ± 1.2 | 104.4 ± 2.2 |
Gen-SH2 | 2.5 μg/L + 0.02 g/L | 121.1 ± 1.2 | 110.9 ± 3.3 |
Gen-SH2 | 2.5 μg/L + 0.05 g/L | 123.1 ± 0.9 | 112.3 ± 3.4 |
Gen-SH2 | 2.5 μg/L + 0.1 g/L | 119.6 ± 1.8 | 107.1 ± 3.9 |
CH1 | 0.02 g/L | 107.6 ± 4.9 | 106.6 ± 5.1 |
CH1 | 0.05 g/L | 108.6 ± 4.4 | 105.9 ± 5.8 |
CH1 | 0.1 g/L | 102.9 ± 6.6 | 105.1 ± 5.6 |
Gen-CH1 | 2.5 μg/L + 0.02 g/L | 108.6 ± 3.3 | 108.1 ± 4.4 |
Gen-CH1 | 2.5 μg/L + 0.05 g/L | 110.5 ± 2.9 | 107.4 ± 1.3 |
Gen-CH1 | 2.5 μg/L + 0.1 g/L | 101.9 ± 4.9 | 104.4 ± 4.6 |
CH2 | 0.02 g/L | 107.9 ± 3.9 | 108.0 ± 3.3 |
CH2 | 0.05 g/L | 112.4 ± 5.1 | 114.6 ± 4.6 |
CH2 | 0.1 g/L | 105.6 ± 1.9 | 113.1 ± 6.7 |
Gen-CH2 | 2.5 μg/L + 0.02 g/L | 107.9 ± 7.0 | 107.3 ± 5.8 |
Gen-CH2 | 2.5 μg/L + 0.05 g/L | 114.6 ± 5.1 | 116.8 ± 5.5 |
Gen-CH2 | 2.5 μg/L + 0.1 g/L | 106.7 ± 3.4 | 113.9 ± 3.8 |
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Wang, S.; Fu, Y.; Zhao, X.-H. The Cooperative Effect of Genistein and Protein Hydrolysates on the Proliferation and Survival of Osteoblastic Cells (hFOB 1.19). Molecules 2016, 21, 1489. https://doi.org/10.3390/molecules21111489
Wang S, Fu Y, Zhao X-H. The Cooperative Effect of Genistein and Protein Hydrolysates on the Proliferation and Survival of Osteoblastic Cells (hFOB 1.19). Molecules. 2016; 21(11):1489. https://doi.org/10.3390/molecules21111489
Chicago/Turabian StyleWang, Shuo, Yu Fu, and Xin-Huai Zhao. 2016. "The Cooperative Effect of Genistein and Protein Hydrolysates on the Proliferation and Survival of Osteoblastic Cells (hFOB 1.19)" Molecules 21, no. 11: 1489. https://doi.org/10.3390/molecules21111489