Chemopreventive Potential of Ethanolic Extracts of Luobuma Leaves (Apocynum venetum L.) in Androgen Insensitive Prostate Cancer
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Extraction
2.3. LC-MS Analysis
2.4. NMR Analysis
2.5. UPLC Analysis
2.6. The Total Phenolic, Flavonoid, Triterpene Contents and Antioxidant Properties
2.7. Cell Culture and Proliferation Assay
2.8. Cell Cycle Analysis
2.9. Assay of Caspase Activity
2.10. Western Blot Analysis
2.11. Statistical Analysis
3. Results
3.1. Extraction, Fractionation, and Cytotoxicity Induced by AVL Ethanolic Extracts
3.2. Identification and Analysis of Bioactive Compounds
3.3. AVL Fraction No. 8 Inhibited Cell Proliferation by G2/M Arrest in PC3 Cells
3.4. AVL F8 Impaired DNA Repair System through Downregulation of the Expression of Uracil-DNA Glycosylase Leading to the Promotion of G2/M Arrest
3.5. Reduced β-Catenin Expression Involved in AVL F8-Induced G2/M Arrest
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Fraction | IC50 (24 h) | Total Flavonoid Content | Total Phenolic Content | Total Triterpenoids | ABTS+ | DPPH |
---|---|---|---|---|---|---|
μg/mL | mg Catecin Equivalents/g | mg Gallic Acid Equivalents/g | Oleanolic Acid (%) | μmol Trolox Equivalents/g | μmol Vitamin C Equivalents/g | |
No. 8 | 99 | 100.3 ± 4.9 | 28.3 ± 1.2 | 51.6 ± 6.3 | 65.2 ± 1.0 | 34.6 ± 0.2 |
No. 10 | 216 | 64.0 ± 5.5 | 35.5 ± 0.1 | 49.4 ± 2.4 | 180.9 ± 3.0 | 55.1 ± 0.1 |
No. 13 | 274 | 25.2 ± 0.8 | 12.8 ± 0.2 | 26.3 ± 1.6 | 118.2 ± 1.6 | 77.4 ± 0.4 |
No. 22 | 240 | 64.0 ± 0.4 | 54.8 ± 0.1 | 79.4 ± 7.5 | 196.1 ± 2.7 | 80.4 ± 0.4 |
No. 25 | 227 | 86.8 ± 0.5 | 51.9 ± 1.2 | 39.4 ± 3.0 | 251.6 ± 3.6 | 96.5 ± 1.4 |
Compounds | In AVL * | In Fract. # | In AVL * | In Fract. # | In AVL * | In Fract. # | In AVL * | In Fract. # | In AVL * | In Fract. # |
---|---|---|---|---|---|---|---|---|---|---|
Fraction | No. 8 | No. 10 | No. 13 | No. 22 | No. 25 | |||||
Sterols | ||||||||||
Sitgmasterol | 5.01 | 4.09 | 10.71 | 25.10 | 0.88 | 1.34 | ||||
β-sitosterol | 1.66 | 3.89 | 19.10 | 29.07 | ||||||
Triterpenoid | ||||||||||
Lupeol | 23.65 | 19.31 | 1.25 | 2.92 | ||||||
Flavonoids | ||||||||||
Kaempferol | 0.37 | 0.60 | 0.12 | 0.36 | ||||||
Isorhamnetin | 0.05 | 0.16 |
Compounds | Molecular Formula | Molecular Weight | Q1 Mass (m/z) |
---|---|---|---|
Sterols | |||
Sitgmasterol | C29H48O | 412.69 | 413 [M + H]+ |
β-sitosterol | C29H50O | 414.71 | 437 [M + Na]+ |
Triterpenoid | |||
Lupeol | C30H50O | 426.72 | 449 [M + Na]+ |
Flavonoids | |||
Kaempferol | C15H10O6 | 286.23 | 287 [M + H]+ |
Isorhamnetin | C16H12O7 | 316.26 | 317 [M + H]+ |
Quercetin | C15H10O7 | 302.24 | 303 [M + H]+ |
Compounds | Selected 1H-NMR Data δ (Multiplicity/Hz) | Selected 13C-NMR Data δ |
---|---|---|
Sterols | ||
Sitgmasterl | 0.70 (3H, s, H-29), 0.82(3H, d, H-26), 0.84 (3H, d, H-24), 0.85 (3H, d, H-27), 1.03 (3H, s, H-28), 3.53 (1H, m, H-3), 5.02 (1H, dd, H-21), 5.16 (1H, dd, H-20), 5.35 (1H, d, H-6) | 12.0 (CH3, C-18), 12.3 (CH3, C-29), 18.8 (CH3, C-21), 19.0 (CH3, C-27), 19.4 (CH3, C-19), 21.0 (CH3, C-26), 21.1 (CH2, C-11), 24.4 (CH2, C-15), 25.4 (CH2, C-23) 28.9 (CH, C-25), 31.6 (CH2, C-2), 31.9 (CH, C-8), 31.9 (CH2, C-7), 36.5 (C, C-10), 37.2 (CH2, C-1), 39.7 (CH2, C-12), 42.2 (CH2, C-4), 42.3 (C, C-13), 50.1 (CH, C-9), 56.0 (CH, C-17), 56.8 (CH, C-14), 71.8 (CH, C-3), 121.7 (CH, C-6), 129.2 (CH, C-3), 138.3 (CH, C-22), 140.7 (C, C-5) |
β-sitosterol | 0.68 (3H, s, H-18), 0.83(3H, d, H-27), 0.84 (3H, d, H-26), 0.90 (3H, s, H-29), 0.93 (3H, d, H-21), 1.01 (3H, s, H-19), 3.54 (1H, m, H-3), 5.35 (1H, dd, H-6) | 1.8 (CH3, C-18), 12.0 (CH3, C-29), 18.8 (CH3, C-21), 19.0 (CH3, C-27), 19.4 (CH3, C-19), 19.8 (CH3, C-26), 21.1 (CH2, C-11), 23.1 (CH2, C-28), 24.3 (CH2, C-15), 26.0 (CH2, C-23), 28.2 (CH2, C-16), 29.1 (CH, C-25), 31.6 (CH2, C-2), 31.8 (CH, C-8), 31.9 (CH2, C-7), 33.9 (CH2, C-22), 36.1 (CH, C-20), 36.5 (C, C-10), 37.2 (CH2, C-1), 39.7 (CH2, C-12), 42.3 (CH2, C-4), 42.3 (C, C-13), 45.8 (CH, C-24), 50.1 (CH, C-9), 56.0 (CH, C-17), 56.7 (CH, C-14), 71.8 (CH, C-3), 121.7 (CH, C-6), 140.7 (C, C-5) |
Triterpenoid | ||
Lupeol | 0.69 (1H, d, H-5), 0.76 (3H, s, H-23), 0.78 (3H, s, H-28), 0.83 (3H, s, H-25), 0.94 (3H, s, H-27), 0.97 (3H, s, H-26), 1.03 (3H, s, H-24), 1.27 (2H, m, H-21), 1.30 (1H, m, H-9), 1.38 (2H, m, H-7), 1.39 (2H, m, H-6), 1.38 (1H, m, H-18), 1.68 (1H, s, H-30), 2.38 (1H, m, H-19), 3.18 (1H, m, H-3), 4.56 (1H, s, H-29b), 4.68 (1H, s, H-29a) | 14.5 (CH3, C-27), 15.4(CH3, C-24), 16.0 (CH3, C-26), 16.1 (CH3, C-25), 18.0 (CH3, C-28), 18.3 (CH2, C-6), 19.3 (CH3, C-30), 20.9 (CH2, C-11), 25.1 (CH2, C-12), 27.2 (CH2, C-15), 27.4 (CH2, C-2), 28.0 (CH3, C-23), 29.8 (CH2, C-21), 34.3 (CH2, C-7), 35.9 (CH2, C-16), 37.2 (C, C-10), 38.0(CH, C-13), 38.7 (CH2, C-1), 38.9 (C, C-4), 40.0 (CH2, C-22), 40.8 (C, C-8), 42.8 (C, C-14), 43.0 (C, C-17), 48.0 (CH, C-19), 48.3 (CH, C-18), 50.4 (CH, C-9), 55.3 (CH, C-5), 79.0 (CH, C-3), 109.3 (CH2, C-29), 151.0 (C, C-20) |
Flavonoids | ||
Kaempferol | 6.20 (1H, d, H-6), 6.44 (1H, d, H-8), 6.93 (1H, d, H-5′), 8.03 (1H, m, H-6′), 9.39 (1H, s, OH), 10.10 (1H, s, OH), 10.78 (1H, s, OH), 12.47 (1H, s, OH) | 146.9 (C-2), 135.7 (C-3), 175.9 (C-4), 156.3 (C-5), 98.3(C-6), 163.9 (C-7), 93.6 (C-8), 160.8(C-9), 103.1 (C-10), 121.7 (C-1′),115.5 (C-2′), 129.6 (C-3′), 159.3 (C-4′), 115.5 (C-5′), 121.7 (C-6′) |
Isorhamnetin | 3.84 (3H, s, OCH3), 6.20 (1H, d, H-6), 6.48 (1H, d, H-8), 6.93 (1H, d, H-5′), 7.70 (1H, m, H-6′), 7.75 (1H, d, H-2′), 9.44 (1H, s, OH), 9.75 (1H, s, OH), 10.76 (1H, s, OH), 12.47 (1H, s, OH) | 148.8 (C-2), 135.8 (C-3), 175.9 (C-4), 156.2 (C-5), 98.2(C-6), 163.9 (C-7), 93.6 (C-8), 160.7(C-9), 103.0 (C-10), 121.9 (C-1′),111.7 (C-2′), 146.6 (C-3′), 147.4 (C-4′),115.5 (C-5′), 121.7 (C-6′), 55.8 (OCH3) |
Quercetin | 6.24 (1H, d, H-6), 6.48 (1H, d, H-8), 6.96 (1H, d, H-5′), 7.63 (1H, m, H-6′), 7.70 (1H, d, H-2′), 12.00 (1H, s, OH) |
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Huang, S.-P.; Ho, T.-M.; Yang, C.-W.; Chang, Y.-J.; Chen, J.-F.; Shaw, N.-S.; Horng, J.-C.; Hsu, S.-L.; Liao, M.-Y.; Wu, L.-C.; et al. Chemopreventive Potential of Ethanolic Extracts of Luobuma Leaves (Apocynum venetum L.) in Androgen Insensitive Prostate Cancer. Nutrients 2017, 9, 948. https://doi.org/10.3390/nu9090948
Huang S-P, Ho T-M, Yang C-W, Chang Y-J, Chen J-F, Shaw N-S, Horng J-C, Hsu S-L, Liao M-Y, Wu L-C, et al. Chemopreventive Potential of Ethanolic Extracts of Luobuma Leaves (Apocynum venetum L.) in Androgen Insensitive Prostate Cancer. Nutrients. 2017; 9(9):948. https://doi.org/10.3390/nu9090948
Chicago/Turabian StyleHuang, Szu-Ping, Tzu-Ming Ho, Chih-Wen Yang, Ya-Ju Chang, Jie-Fu Chen, Ning-Sing Shaw, Jia-Cherng Horng, Shih-Lan Hsu, Ming-Yuan Liao, Li-Chen Wu, and et al. 2017. "Chemopreventive Potential of Ethanolic Extracts of Luobuma Leaves (Apocynum venetum L.) in Androgen Insensitive Prostate Cancer" Nutrients 9, no. 9: 948. https://doi.org/10.3390/nu9090948