Background
Methods
Chemicals and reagents
Plant materials and preparation of extracts
Quantitative determination of the major components of plant extracts by HPLC
Validation procedures for HPLC analysis
Isolation of human polymorphonuclear leukocytes and monocytes
Cell viability
Chemiluminescence assay
Chemotaxis assay
Phagocytic assay
CD18 integrin expression of leukocytes
Statistical analysis
Results and discussion
Species | Phyllanthin | Hypophyllanthin | Gallic acid | Ellagic acid | Corilagin | Geraniin |
---|---|---|---|---|---|---|
PA (Mal) | 103.50 | 114.00 | 163.30 | 601.29 | 313.41 | 170.49 |
PA (Ind) | 79.10 | 121.85 | 142.92 | 219.06 | 413.42 | 234.52 |
PU (Mal) | 16.80 | 19.50 | 103.48 | 231.34 | 159.37 | 173.17 |
PU (Ind) | 1.9 | 3.91 | 93.05 | 346.12 | 379.23 | 199.50 |
Chemotaxis assay
Sample | Chemotaxis | Chemiluminescence | ||||||
---|---|---|---|---|---|---|---|---|
fMLP | Zymosan | PMA | ||||||
PMNs | MNCs | Whole Blood | PMNs | MNCs | Whole Blood | PMNs | MNCs | |
PA (Mal) | 1.22 ± 0.10 | 1.44 ± 0.80 | 0.95 ± 0.24 | 0.58 ± 1.80 | 0.10 ± 0.17 | 1.24 ± 0.98 | 0.42 ± 0.75 | 0.56 ± 0.30 |
PA (Ind) | 1.84 ± 0.01 | 2.35 ± 0.23 | 8.14 ± 1.71 | 0.12 ± 0.05 | 2.19 ± 2.55 | 6.02 ± 1.33 | 1.34 ± 0.59 | 1.27 ± 0.80 |
PU (Mal) | 1.46 ± 0.10 | 2.84 ± 0.30 | 1.52 ± 0.53 | 1.00 ± 1.21 | 1.23 ± 0.88 | 1.98 ± 1.09 | 0.45 ± 0.23 | 0.26 ± 0.12 |
PU (Ind) | 2.21 ± 0.90 | 2.92 ± 0.14 | 9.74 ± 1.09 | 1.64 ± 0.58 | 0.36 ± 0.29 | 4.80 ± 1.80 | 1.34 ± 1.18 | 1.06 ± 0.31 |
GA | 1.40 ± 0.02 (8.22 ± 0.15) | 2.00 ± 0.03 (11.75 ± 0.23) | 1.70 ± 0.03 (9.99 ± 0.12) | 0.43 ± 1.02 (2.53 ± 0.10) | 1.29 ± 0.87 (7.58 ± 0.50) | 0.87 ± 0.12 (5.11 ± 0.15) | 0.48 ± 0.01 (2.82 ± 0.15) | 1.28 ± 0.01 (7.52 ± 0.12) |
EA | 1.33 ± 0.07 (4.40 ± 0.12) | 1.81 ± 0.12 (5.96 ± 0.12) | 1.36 ± 0.08 (4.50 ± 0.16) | 0.56 ± 0.02 (1.85 ± 0.35) | 0.45 ± 0.09 (1.49 ± 0.07) | 0.83 ± 0.03 (2.75 ± 0.07) | 0.31 ± 0.07 (1.02 ± 0.06 ) | 0.75 ± 0.12 (2.48 ± 0.06 ) |
Ger | 1.04 ± 0.12 (1.09 ± 0.23 ) | 1.61 ± 0.14 (1.69 ± 0.05) | 0.34 ± 0.30 (0.36 ± 0.12 ) | 0.18 ± 0.07 (0.18 ± 0.02) | 0.37 ± 0.41 (0.38 ± 0.07) | 0.19 ± 0.12 (0.21 ± 0.01) | 0.12 ± 0.04 (0.13 ± 0.01) | 0.36 ± 0.61 (0.38 ± 0.12 ) |
Cor | 1.49 ± 0.024 (2.34 ± 0.07) | 1.99 ± 0.09 (3.13 ± 0.44) | 0.37 ± 0.37 (0.58 ± 0.03) | 0.36 ± 0.07 (0.56 ± 0.023) | 0.44 ± 0.08 (0.69 ± 0.12) | 0.22 ± 0.88 (0.35 ± 0.09) | 0.15 ± 0.04 (0.24 ± 0.02) | 0.44 ± 0.03 (0.69 ± 0.05) |
Phyll | 3.20 ± 0.15 (7.60 ± 0.10) | 8.51 ± 0.52 (20.2 ± 0.40) | 3.70 ± 2.80 (8.80 ± 1.00) | 3.2 ± 2.81 (7.6 ± 0.40) | 2.38 ± 1.44 (5.68 ± 0.31) | 1.40 ± 1.72 (3.34 ± 0.05) | 0.08 ± 0.66 (0.20 ± 0.01) | 1.54 ± 0.87 (3.68 ± 0.43) |
Hypophyll | 4.40 ± 0.2 (10.2 ± 0.40) | 9.60 ± 0.26 (22.3 ± 0.53) | 3.6 ± 1.40 (8.4 ± 1.30) | 6.1 ± 2.32 (14.2 ± 1.70) | 1.41 ± 0.06 (3.27 ± 0.15) | 1.87 ± 0.01 (4.34 ± 0.02) | 0.95 ± 0.11 (2.20 ± 0.31) | 1.73 ± 0.31 (4.02 ± 0.71) |
Ibuprofen | 1.40 ± 0.10 (6.60 ± 0.10) | 1.98 ± 0.05 (9.57 ± 0.23) | ||||||
Aspirin | 2.16 ± 0.80 (11.88 ± 4.14) | 0.85 ± 0.20 (4.68 ± 2.71) | 0.43 ± 0.07 (2.40 ± 0.16) | 0.17 ± 0.01 (0.94 ± 0.05) | 0.14 ± 0.09 (0.77 ± 0.16) | 0.58 ± 0.28 (3.22 ± 0.38) |
Phagocytic assay
Sample (μg/mL) | Neutrophils | Monocytes | ||||||
---|---|---|---|---|---|---|---|---|
100 | 50 | 6.25 | 3.125 | 100 | 50 | 6.25 | 3.125 | |
PA (Mal) | 75.60 ± 1.82 | 88.20 ± 1.17 | 53.70 ± 0.15 | 65.68 ± 0.15 | ||||
PA (Ind) | 81.65 ± 2.21 | 89.29 ± 2.67 | 66.68 ± 3.16 | 74.60 ± 2.76 | ||||
PU (Mal) | 78.13 ± 2.07 | 89.60 ± 0.98 | 59.08 ± 1.66 | 68.80 ± 1.66 | ||||
PU (Ind) | 73.30 ± 3.32 | 79.40 ± 1.07 | 61.86 ± 2.81 | 72.22 ± 3.02 | ||||
GA | 88.60 ± 1.52 | 95.90 ± 1.97 | 76.50 ± 2.18 | 77.80 ± 1.71 | ||||
EA | 85.20 ± 1.19 | 88.18 ± 2.13 | 75.50 ± 1.97 | 67.10 ± 2.31 | ||||
Ger | 76.00 ± 3.05 | 80.80 ± 1.06 | 76.08 ± 3.16 | 84.90 ± 0.91 | ||||
Cor | 85.50 ± 2.82 | 86.10 ± 0.32 | 80.70 ± 1.51 | 85.38 ± 1.04 | ||||
Phyll | 14.20 ± 1.08 | 34.91 ± 0.09 | 27.10 ± 0.55 | 52.10 ± 0.68 | ||||
Hypophyll | 49.11 ± 1.04 | 80.50 ± 0.87 | 64.6 ± 0.51 | 67.0 ± 1.07 | ||||
Positive control | 91.60 ± 1.15 | 84.6 ± 4.84 |
Inhibition of CD18 expression assay
Sample (μg/mL) | Neutrophils | Monocytes | ||||||
---|---|---|---|---|---|---|---|---|
100 | 50 | 6.25 | 3.125 | 100 | 50 | 6.25 | 3.125 | |
PA (Mal) | 79.62 ± 1.82 | 87.10 ± 1.23 | 70.79 ± 2.13 | 85.68 ± 1.10 | ||||
PA (Ind) | 81.65 ± 2.21 | 89.29 ± 2.67 | 62.39 ± 1.87 | 89.29 ± 1.51 | ||||
PU (Mal) | 82.90 ± 1.08 | 90.62 ± 1.80 | 76.20 ± 1.54 | 88.98 ± 2.76 | ||||
PU (Ind) | 87.15 ± 3.39 | 93.20 ± 3.39 | 85.18 ± 0.51 | 85.41 ± 1.69 | ||||
GA | 81.33 ± 1.80 | 88.77 ± 2.41 | 68.60 ± 1.20 | 76.50 ± 2.18 | ||||
EA | 86.14 ± 2.01 | 90.18 ± 1.07 | 62.20 ± 0.57 | 77.10 ± 1.99 | ||||
Ger | 89.40 ± 1.21 | 93.10 ± 1.23 | 82.90 ± 1.71 | 88.40 ± 1.75 | ||||
Cor | 92.10 ± 1.02 | 95.80 ± 0.98 | 79.51 ± 1.10 | 85.38 ± 1.32 | ||||
Phyll | 67.17 ± 1.34 | 79.20 ± 2.92 | 61.56 ± 0.98 | 72.21 ± 1.26 | ||||
Hypophyll | 74.70 ± 0.09 | 80.90 ± 1.65 | 74.42 ± 0.77 | 75.15 ± 1.48 | ||||
Positive control | 93.93 ± 0.54 | 84.79 ± 0.55 |