B Type and Complex A/B Type Epicatechin Trimers Isolated from Litchi pericarp Aqueous Extract Show High Antioxidant and Anticancer Activity
Abstract
:1. Introduction
2. Results and Discussion
2.1. A Fraction from Litchi phenolic Extract Displayed Strong Inhibition on Cancer Cell Proliferation and Antioxidant Activity
2.2. The Fraction with High Anti-Cancer Activity was Further Separated, and a Sub-Fraction S3 Was Isolated with High Activity
2.3. The Sub-Fraction with High Anti-Cancer Activity Was Further Separated and a Fraction Contained a B-Type Trimer Proanthocyanidin Showed High Antioxidant and Anticancer Activities
2.4. Contents of Catechins and Proanthocyanidins in Litchi pericarp during Fruit Development
2.5. Expression of Catechin and Anthocyanin Biosynthesis Related Genes in Litchi pericarp during Fruit Development
3. Materials and Methods
3.1. Plant Materials
3.2. Extraction and Purification of Litchi pericarp Phenolics
3.3. Analysis for Total Phenolic Compounds
3.4. Determination of Total Flavonoid Content
3.5. Ferric Reducing Antioxidant Power (FRAP) Assays
3.6. DPPH Radical Scavenging Activity
3.7. Inhibition Activity Assay on Cancer Cells
3.8. HPLC-ESI-MS/MS Analysis of Fractions P1–P4
3.9. Determination of the Contents of Catechin-Type Compounds and Proanthocyanidins by HPLC
3.10. Gene Expression Analysis
3.11. Statistics
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
GAE | gallic acid equivalents |
RE | rutin equivalents |
CCK-8 | cell counting Kit-8 |
EC | (−)-epicatechin |
C | (+)-catechin |
EGCG: | (−)-epigallocatechin gallate; |
ECG | (−)-epicatechin-3-gallate |
CG | (−)-catechin gallate |
FRAP | Ferric reducing antioxidant power |
DAA | day after anthesis |
CHI | chalcone isomerase |
ANS | anthocyanin synthase |
UFGT | UDP-glucose: flavonoid 3-O-glucosyltransferase |
ANR | anthocyanidin reductase |
LAR | leuacoanthocyanidin reductase |
SEM | standard error of mean |
BHT | butylated hydroxytoluene |
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Peak | Compound Number | Mode | Mass | m/z | * Molecular Formula | Auto MS-MS Fragment Ions | * Tentative Identification |
---|---|---|---|---|---|---|---|
P1 | 1 | pos | 290.0792 | 291.0869 | C15H14O6 | 139.0388; 123.044 | EC (Epicatechin) [20,21] |
neg | 290.1537 | 289.1464 | 245.0816; 125.0243 | ||||
P2 | 2 | pos | 578.1425 | 579.1498 | C30H26O12 | 127.0391; 291.0866; 409.0920 | PA (Proanthocyanidin) B2 [22] |
neg | 578.1418 | 577.1345 | 125.0244; 289.0713; 407.0778 | ||||
P3 | 3 | pos | 864.1899 | 887.1792 | C45H36O18 | 289.0714; 425.0902; 577.1345; 713.1506 | Pavetannin B2 (Trimers of complex Proanthocyanidin A and B) [23] |
neg | 864.1899 | 863.1867 | 125.0244; 289.0717; 423.0712; 575.1186; 711.1357 | ||||
4 | pos | 866.2053 | 889.1945 | C45H38O18 | 289.0706; 601.1310; 717.1510 | Litchitannin B1 (Trimers of Proanthocyanidin B) | |
neg | - | - | - | ||||
P4 | 5 | pos | - | - | C30H24O12 | - | 2 × PA(A) (Dimers of Proanthocyanidin A) [24] |
neg | 576.1257 | 575.1184 | - | ||||
6 | pos | 866.2056 | 867.2129 | C45H38O18 | 247.0601; 289.0705; 427.1025; 579.1492; 715.1647 | Litchitannin B2 (Trimers of Proanthocyanidin B) | |
neg | 866.2053 | 865.1981 | 125.0243; 287.0556; 407.0755; 575.1187; 713.1531 | ||||
7 | pos | 864.1902 | 865.1974 | C45H36O18 | 287.0550; 411.1073; 533.1076; 575.1179; 713.1497 | 3 × PA(A&B) (Trimers of complex Proanthocyanidin A and B) [23] | |
neg | 864.1899 | 863.1826 | 125.0247; 289.0716; 411.0720; 573.1025; 711.1346 | ||||
8 | pos | 848.1951 | 849.2023 | C45H36O17 | 287.0557; 517.1125; 713.1504 | Trimeric PA (with an afzelechin or epiafzelechin) [23] | |
neg | 848.1948 | 847.1875 | 289.0710; 411.0723; 557.1095 | ||||
9 | pos | 864.1905 | 865.1979 | C45H36O18 | 287.0551; 425.0869; 577.1336; 713.1497 | 3 × PA (A&B) [23] | |
neg | 864.1899 | 863.1827 | 125.0243; 285.0404; 575.1201; 711.1362 | ||||
10 | pos | 576.127 | 577.1342 | C30H24O12 | 123.0440; 287.0549; 425.0864 | Procyanidin A2 [25] |
Genes | GenBank ID | Forward Primer (5′ to 3′) | Reverse Primer (5′ to 3′) |
---|---|---|---|
LcCHI | HQ402910 | CGGAGTTTACTTGGAGGATGT | CAGTGACCTTCTCAGAGTATTG |
LcANS | HQ402913 | AGGAAGTTGGTGGTCTGGAAG | CCGTTGCTGAGGATTTCAATGGTG |
LcUFGT | HQ402914 | GCCACCAGCGGTTCCTAATA | ATGCCTCTGCTACTGCTACAATCT |
LcLAR | Litchi_GLEAN_10054331 | AGATTGGACGAACTCTACC | TTGAAACAATCATCCAACG |
LcANR | Litchi_GLEAN_10049142 | ATACATTTGTGCTGCTTTC | TCATAGGTTTCTTCAAGCC |
LcActin | HQ615689 | ACCGTATGAGCAAGGAAATCACTG | TCGTCGTACTCACCCTTTGAAATC |
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Gong, Y.; Fang, F.; Zhang, X.; Liu, B.; Luo, H.; Li, Z.; Zhang, X.; Zhang, Z.; Pang, X. B Type and Complex A/B Type Epicatechin Trimers Isolated from Litchi pericarp Aqueous Extract Show High Antioxidant and Anticancer Activity. Int. J. Mol. Sci. 2018, 19, 301. https://doi.org/10.3390/ijms19010301
Gong Y, Fang F, Zhang X, Liu B, Luo H, Li Z, Zhang X, Zhang Z, Pang X. B Type and Complex A/B Type Epicatechin Trimers Isolated from Litchi pericarp Aqueous Extract Show High Antioxidant and Anticancer Activity. International Journal of Molecular Sciences. 2018; 19(1):301. https://doi.org/10.3390/ijms19010301
Chicago/Turabian StyleGong, Yihui, Fang Fang, Xin Zhang, Bin Liu, Honghui Luo, Zhen Li, Xuelian Zhang, Zhaoqi Zhang, and Xuequn Pang. 2018. "B Type and Complex A/B Type Epicatechin Trimers Isolated from Litchi pericarp Aqueous Extract Show High Antioxidant and Anticancer Activity" International Journal of Molecular Sciences 19, no. 1: 301. https://doi.org/10.3390/ijms19010301