Screening In Vitro Targets Related to Diabetes in Herbal Extracts from Peru: Identification of Active Compounds in Hypericum laricifolium Juss. by Offline High-Performance Liquid Chromatography
Abstract
:1. Introduction
2. Results and Discussion
2.1. Evaluation of α-Glucosidase and Aldose Reductase (AR) Inhibition and Antioxidant Activity of Peruvian Plants
2.2. Effect of Hypericum laricifolium Juss. (HL) on α-Glucosidase and Aldose Reductase (AR) Inhibition and Antioxidant Activity
2.3. Identification of the Major Bioactive Components in Hypericum laricifolium Juss. (HL) by Offline High-Performance Liquid Chromatography (HPLC)
2.3.1. Identification of the Bioactive Compounds in HL by α-Glucosidase Ultrafiltration Combined with HPLC
2.3.2. Identification of the Bioactive Compounds in HL by Aldose Reductase (AR) and Human Recombinant AR (HRAR) Ultrafiltration Combined with HPLC
2.3.3. Identification of Antioxidants Using Offline 2,2-Diphenyl-1-Picrylhydrazyl (DPPH)-(HPLC) and 2,2′-Azino-Bis (3-Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS)-(HPLC)
3. Materials and Methods
3.1. Materials
3.2. Collection of Plant Material
3.3. Preparation of Extracts and Isolation of Plant Samples
3.4. Evaluation of the α-Glucosidase Inhibitory Assay
3.5. Animal Care
3.6. Evaluation of the Rat Lens Aldose Reductase (RLAR) Assay
3.7. Evaluation of the DPPH Assay
3.8. Evaluation of the ABTS Assay
3.9. Determination of the α-Glucosidase Ultrafiltration-HPLC Assay
3.10. Determination of the HRAR Ultrafiltration-HPLC Assay
3.11. Evaluation of Offline DPPH-HPLC Analysis
3.12. Evaluation of Offline ABTS-HPLC Analysis
3.13. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
AR | Aldose reductase |
DPPH | 2,2-Diphenyl-1-picrylhydrazyl |
ABTS | 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) |
DMSO | Dimethyl sulfoxide |
HL | Hypericum laricifolium Juss. |
MeOH | Methanol |
RLAR | Rat lens aldose reductase |
HRAR | Human recombinant aldose reductase |
HPLC | High-performance liquid chromatography |
TBD | Total binding degree |
QR | Quantitative reduction |
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No. | Voucher | Scientific Name | Common Name | Family | Used Part 1 | References
[13,17,19,20,21,22,23,24,25,26,27,28] 2 | Yield (%) 3 | Inhibition (%) 4 | |||
---|---|---|---|---|---|---|---|---|---|---|---|
α-Glucosidase (500 μg/mL) 5 | AR (10 μg/mL) 6 | Antioxidants | |||||||||
DPPH (143 μg/mL) 7 | ABTS (33 μg/mL) 8 | ||||||||||
1 | A5 | Annona muricata L. | Hoja de huanabana, Graviola | ANNONACEAE | L | Hypertension [ 24], Inflammation [20,24] | 19.5 | 26.3 ± 1.7 c | 0.8 ± 0.8 a | 64.2 ± 0.9 e,f,g,h,i | 31.5 ± 2.9 f,g |
2 | A32 | Acanthoxanthium spinosu (L.) Fourr. | Juan Alonso | ASTERACEAE | A | Diabetes [ 25], Inflammation [20] | 8.4 | 3.2 ± 2.7 a | 22.4 ± 1.2 e,f | 11.0 ± 3.5 a,b | 2.1 ± 1.8 a |
3 | A16 | Ambrosia arborescens Mill. | Marco | ASTERACEAE | L | Inflammation [ 25] | 12.7 | 18.5 ± 1.9 c | 31.8 ± 6.1 f,g | 3.5 ± 0.7 a,b | 6.1 ± 1.8 a,b |
4 | P78 | Baccharis genistelloides (Lam.) Pers. | Karqueja | ASTERACEAE | A | Diabetes [ 20,21], Inflammation [20], Burn fat [20], Cholesterol [20,21] | 19.7 | 6.2 ± 3.4 a,b | NA | 47.0 ± 9.2 c,d | 27.9 ± 1.1 e,f,g |
5 | P77 | Chuquiraga spinosa Less. | Huamanpinta | ASTERACEAE | A | Inflammation [ 25], Kidneys [25] | 28.7 | NA 9 | 12.2 ± 0.7 c,d | 70.3 ± 11.0 g,h,i,j | 50.3 ± 3.1 h |
6 | A31 | Flaveria Bidentis (L.) Kuntze | Mata gusano | ASTERACEAE | L | Inflammation [ 26] | 7.9 | 17.3 ± 8.4 b,c | 86.4 ± 1.9 k | 5.3 ± 10.8 a,b | 8.0 ± 2.1 a,b |
7 | P14 | Schkuhria pinnata (Lam.) Kuntze ex Thell. | Canchalagua | ASTERACEAE | A | Diabetes [ 20,21], Inflammation [20,21] | 2.4 | 0.1 ± 2.7 a | NA | 67.1 ± 3.3 f,g,h,i | 71.4 ± 8.4 j |
8 | A6 | Smallanthus sonchifolius (Poepp.) H. Rob. | Hojas de yacon | ASTERACEAE | L | Diabetes [ 20,21,22,23], Inflammation [20], Kidneys [20], Cholesterol [20] | 5.3 | 5.5 ± 1.4 a,b | 31.9 ± 9.1 g | NA | 5.5 ± 3.0 a,b |
9 | P49 | Taraxacum officinale F.H. Wigg. | Diente de leon | ASTERACEAE | A, F | Inflammation [ 20] | 4.8 | NA | NA | 41.9 ± 10.8 c | 15.7 ± 1.6 b,c,d |
10 | A17 | Tessaria integrifolia Ruiz & Pav. | Pajaro bobo | ASTERACEAE | L | Kidneys [ 20], Liver [20], Inflammation [20] | 18.4 | 41.6 ± 1.1 d | 79.9 ± 0.1 k | 52.2 ± 3.9 c,d,e | 31.1 ± 3.3 f,g |
11 | P79 | Cordia lutea Lam. | Flor de overo | BORAGINACEAE | F | Inflammation [ 20,21], Liver [20,21] | 12.3 | NA | 3.6 ± 1.3 a,b | 60.2 ± 2.1 d,e,f,g,h | 36.6 ± 1.7 g |
12 | P36 | Tiquilia Paronychioides (Phil.) Rich. | Flor de arena | BORAGINACEAE | A | Inflammation [ 20] | 18.5 | 2.9 ± 1.1 a | 26.0 ± 0.1 f,g | 95.7 ± 0.8 m,n | 97.1 ± 0.4 m |
13 | A24 | Sambucus peruviana H. B. K. | Sauco | CAPRIFOLIAEAE | L | Kidneys [ 20,23], Inflammation [20] | 8.1 | 47.4 ± 4.8 d | 47.2 ± 0.9 h | 53.3 ± 1.5 c,d,e,f | 33.8 ± 6.1 g |
14 | A10 | Hypericum laricifolium Juss. | Hierba de la fortuna | CLUSIACEAAE | L | Inflammation [ 13,28], Infections, Musculoskeletal, Bone pain [19] | 15.9 | 97.2 ± 2.0 h | 56.9 ± 5.6 i | 81.9 ± 2.5 j,k,l,m | 58.8 ± 4.6 h,i |
15 | P55 | Equisetum giganteum L. | Cola de caballo | EQUISETACEAE | A | Kidneys [ 20,21], Inflammation [20,21] | 11.2 | 77.8 ± 5.9 g | 0.7 ± 0.0 a | 73.9 ± 8.9 h,i,j,k | 51.8 ± 0.3 h |
16 | P5 | Phyllanthus niruri L. | Chanca piedra | EUPHORBIACEAE | L | Diabetes [ 24], Kidneys [20], Liver [20], Inflammation [20] | 12.9 | 39.9 ± 1.4 d | 25.3 ± 3.7 f,g | 97.0 ± 0.1 n | 99.6 ± 0.1 m |
17 | P80 | Desmodium molliculum (Kunth) A.P. De Candolle | Manayupa | FABACEAE | L | Kidneys [ 20,21], Inflammation [20,21] | 23.8 | 97.9 ± 9.1 h | 10.9 ± 0.5 b,c,d | 89.9 ± 4.2 l,m,n | 73.6 ± 1.3 j,k |
18 | A3 | Otholobium mexicanum (L. f.) J.W. Grimes | Culen negro | FABACEAE | A | Diabetes [ 21] | 6.1 | NA | 50.4 ± 0.5 h,i | 4.5 ± 5.7 a,b | 9.6 ± 1.9 a,b,c |
19 | A4 | Otholobium pubescens (Poir.) J.W. Grimes | Culen blanco | FABACEAE | A | Diabetes [ 21] | 19.6 | NA | 8.8 ± 0.5 a,b,c,d | 6.1 ± 0.5 a,b,c,d | 27.4 ± 1.5 e,f,g |
20 | A49 | Vicia Faba | Haba | FABACEAE | Fr | Renal disorders [ 22] | 7.6 | 0.1 ± 2.7 a | NA | 4.2 ± 2.0 a,b | 13.0 ± 3.0 b,c,d |
21 | P7 | Gentianella tristicha (Gilg) J.S. Pringle | Hercampure | GENTIANACEAE | A | Diabetes [ 17], Cholesterol [26] | 30.3 | NA | 68.1 ± 4.2 j | 53.1 ± 0.6 cdef | 66.5 ± 4.5 i,j |
22 | A35 | Geranium dielsianum R. Knuth | Pasuchaca | GERANIACEAE | L | Diabetes [ 17,24] | 6.3 | 97.7 ± 1.1 h | 15.6 ± 0.2 d,e | 96.8 ± 0.3 n | 83.9 ± 4.1 k,l |
23 | P11 | Clinopodium brevicalyx (Epling) Harley & A. Granda | Inka muña | LAMIACEAE | L | Inflammation [ 26] | 26.8 | 1.8 ± 5.5 a | 31.9 ± 0.7 g | 95.8 ± 0.2 m,n | 94.1 ± 2.7 l,m |
24 | P4 | Salvia hispanica L. | Chia | LAMIACEAE | S | Diabetes [ 27], Obesity [27] | 3.5 | NA | NA | 17.3 ± 3.0 b | 21.7 ± 1.8 d,e,f |
25 | P40 | Peumus boldus Molina | Boldo | MONIMIACEAE | L | Inflammation [ 20], Kidneys [20], Liver [20] | 32.5 | 75.2 ± 3.0 g | 7.6 ± 0.9 a,b,c,d | 86.2 ± 3.9 k,l,m,n | 95.7 ± 0.5 m |
26 | P39 | Eucalyptus globolus Labill. | Eucalipto | MYRTACEAE | L | Burn fat [ 20] | 18.0 | 62.3 ± 0.9 e,f | 11.2 ± 0.8 b,c,d | 77.5 ± 0 i,j,k,l | 99.5 ± 0.3 m |
27 | A2 | Argyrochosma nivea (Poir.) Windham | Cuti - Cuti hembra blanca | PTERIDACEAE | A | Diabetes [ 17] | 2.3 | 50.5 ± 3.2 d,e | 59.1 ± 0.3 i,j | 71.2 ± 0.7 g,h,i,j | 34.7 ± 7.3 g |
28 | P17 | Cheilanthes pilosa Goldm. | Cuti Cuti | PTERIDACEAE | A | Diabetes [ 26], Liver [26] | 14.3 | 72.4 ± 0.6 f,g | 9.6 ± 0.9 b,c,d | 58.9 ± 4.8 d,e,f,g | 56.4 ± 4.3 h,i |
29 | A1 | Cheilanthes pruinata Kaulf. | Cuti - Cuti marron macho | PTERIDACEAE | A | Diabetes [ 17] | 30.3 | 73.4 ± 2.9 f,g | 41.6 ± 1.1 h | 53.2 ± 0.3 c,d,e,f | 31.1 ± 4.1 f,g |
30 | A19 | Buddleja Americana L. | Flor blanca | SCROPHULARIA CEAE | F | Inflammation [ 26] | 8.5 | 16.1 ± 5.6 b,c | 3.7 ± 0.1 a,b,c | 2.4 ± 1.0 a | 19.6 ± 1.2 c,d,e |
Extracts | α-Glucosidase 1 | AR 2 | DPPH 3 | ABTS 4 | ||||
---|---|---|---|---|---|---|---|---|
Inhibition (%) | IC50 (μg/mL) 5 | Inhibition (%) | IC50 (μg/mL) | Inhibition (%) | IC50 (μg/mL) | Inhibition (%) | IC50 (μg/mL) | |
MC | 11.65 ± 2.1 * | - | 28.9 ± 7.4 * | - | 17.5 ± 0.3 * | - | 14.4 ± 1.3 * | - |
70% MeOH | 92.36 ± 1.1 * | 56.6 ± 2.5 | 64.51 ± 1.3 * | 3.3 ± 52 | 93.0 ± 0.1 * | 42.5 ± 0.6 | 78.9 ± 0.6 * | 14.4 ± 1.3 |
Acarbose | 55.82 ± 2.3 | 367.4 ± 2.1 | - | - | - | - | - | - |
Quercetin | - | - | 83.7 ± 2.6 | 1.3 ± 3.6 | - | - | - | - |
L-Ascorbic | - | - | - | - | 99.1 ± 0 | 17.6 ± 0.1 | - | - |
Trolox | - | - | - | - | - | - | 100 ± 0.1 | 4.6 ± 0.1 |
Compounds | α-glucosidase 1 | AR 2 | Antioxidants | |||||
---|---|---|---|---|---|---|---|---|
DPPH 3 | ABTS 4 | |||||||
Conc. (μg/mL) | IC50 (μM) 5 | Conc. (μg/mL) | IC50 (μM) | Conc. (μg/mL) | IC50 (μM) | Conc. (μg/mL) | IC50 (μM) | |
Protocatechuic acid (1) | 50 | NA 6 | 10 | 16.9 ± 1.9 e | 143 | 263.4 ± 0.2 e | 17 | 9.7 ± 0.9 c |
p-Hydroxybenzoic acid (2) | 50 | NA | 10 | NA | 143 | NA | 33 | NA |
Chlorogenic acid (3) | 50 | NA | 10 | 0.23 ± 1.8 a | 143 | 123.1 ± 2.9 d | 33 | 23.1 ± 3.5 f |
Vanilic acid (4) | 50 | NA | 10 | NA | 143 | NA | 33 | NA |
Caffeic acid (5) | 50 | NA | 10 | 28.3 ± 0.9 f | 143 | 47.2 ± 0.1 b | 17 | 7.2 ± 1.1 b |
Kaempferol 3-O-glucuronide (6) | 50 | NA | 10 | 6.0 ± 1.1 c | 143 | 58.8 ± 2.1 c | 17 | 11.0 ± 0.2 d |
Quercetin (7) | 50 | 15.9 ± 1.2 a | 10 | 2.5 ± 0.8 b | 71 | 0.33 ± 1.4 a | 17 | 0.33 ± 0.6 a |
Kaempferol (8) | 50 | NA | 10 | 9.7 ± 0.5 d | 143 | 326.4 ± 3.3 f | 33 | 191.8 ± 1.1 g |
Acarbose | 500 | 439.9 ± 8.9 b | - | - | - | - | - | - |
Quercetin | - | - | 10 | 2.4 ± 1.2 b | - | - | - | - |
l-Ascorbic | - | - | - | - | 71 | 0.57 ± 0.4 a | - | - |
Trolox | - | - | - | - | - | - | 17 | 18.8 ± 0.5 e |
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Guillen Quispe, Y.N.; Hwang, S.H.; Wang, Z.; Zuo, G.; Lim, S.S. Screening In Vitro Targets Related to Diabetes in Herbal Extracts from Peru: Identification of Active Compounds in Hypericum laricifolium Juss. by Offline High-Performance Liquid Chromatography. Int. J. Mol. Sci. 2017, 18, 2512. https://doi.org/10.3390/ijms18122512
Guillen Quispe YN, Hwang SH, Wang Z, Zuo G, Lim SS. Screening In Vitro Targets Related to Diabetes in Herbal Extracts from Peru: Identification of Active Compounds in Hypericum laricifolium Juss. by Offline High-Performance Liquid Chromatography. International Journal of Molecular Sciences. 2017; 18(12):2512. https://doi.org/10.3390/ijms18122512
Chicago/Turabian StyleGuillen Quispe, Yanymee N., Seung Hwan Hwang, Zhiqiang Wang, Guanglei Zuo, and Soon Sung Lim. 2017. "Screening In Vitro Targets Related to Diabetes in Herbal Extracts from Peru: Identification of Active Compounds in Hypericum laricifolium Juss. by Offline High-Performance Liquid Chromatography" International Journal of Molecular Sciences 18, no. 12: 2512. https://doi.org/10.3390/ijms18122512