Background
Materials and methods
Reagents and materials
Preparation of Radix isatidis protein
Analysis of the Radix isatidis protein
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis
Amino acid analysis
Evaluation of nutritional parameters
Determination of the antioxidant activity of RIP in vivo
Animal selection and experimental design
Sample preparation of serum, kidney, liver and brain tissue
Histological analysis
Analysis of kidney, liver, thymus and spleen indices
Biochemical analysis
Statistical analysis
Results
SDS-PAGE analysis
The amino acids composition of RIP
Amino acids | RIP | FAO/WHO suggested requirements (2–5 years old) |
---|---|---|
Isoleucine | 7.36 ± 0.51 | 2.8 |
Leucine | 3.92 ± 0.24 | 6.6 |
Lysine | 0.46 ± 0.06 | 5.8 |
Cysteine | 0.01 ± 0.00 | |
Methionine | 0.11 ± 0.02 | |
Total sulphur amino acids | 0.12 | 2.5 |
Tyrosine | 0.85 ± 0.06 | |
Phenylalanine | 4.08 ± 0.19 | |
Total aromatic amino acids | 4.93 | 6.3 |
Threonine | 5.06 ± 0.83 | 3.4 |
Tryptophan | 2.57 ± 0.44 | 1.1 |
Valine | 7.62 ± 0.08 | 3.5 |
Histidine | 1.74 ± 0.04 | 1.9 |
Total essential amino acids | 33.79 | 32.8 |
Arginine | 2.95 ± 0.24 | |
Aspartic acid + asparagine | 9.78 ± 0.65 | |
Glutamic acid + glutamine | 6.03 ± 0.42 | |
Serine | 2.36 ± 0.04 | |
Proline | 8.88 ± 0.49 | |
Glycine | 6.03 ± 0.70 | |
Alanine | 2.43 ± 0.21 | |
Total non-essential amino acids | 38.46 | |
aE/T,% | 46.77 | |
Amino acid score | 101.33 | |
bAntioxidant amino acids | 29.63 |
Body weight
Group | Week | |||||||
---|---|---|---|---|---|---|---|---|
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | |
Normal control | 21.26 ± 1.18 | 26.60 ± 1.57 | 29.50 ± 1.02 | 32.32 ± 1.30 | 36.33 ± 1.25 | 38.34 ± 1.57 | 40.08 ± 1.60 | 42.77 ± 1.15 |
Model control | 21.49 ± 1.24 | 26.59 ± 2.62 | 29.74 ± 1.28 | 32.40 ± 1.14 | 36.51 ± 1.32 | 35.68 ± 1.65d | 35.04 ± 0.92d | 34.19 ± 1.16d |
Positive control | 21.38 ± 1.05 | 26.77 ± 1.11 | 28.41 ± 1.67 | 32.33 ± 2.23 | 35.99 ± 2.62 | 36.54 ± 1.59ac | 39.02 ± 2.39b | 41.59 ± 2.92b |
RIP (25 mg/kg) | 21.26 ± 0.75 | 26.08 ± 1.43 | 27.67 ± 1.24ac | 31.08 ± 1.57 | 33.86 ± 2.47ac | 36.14 ± 2.42ac | 38.00 ± 2.84bc | 40.65 ± 1.25b |
RIP (50 mg/kg) | 21.56 ± 0.69 | 26.26 ± 0.85 | 30.85 ± 1.27 | 32.18 ± 0.84 | 35.54 ± 2.38 | 37.83 ± 2.89c | 38.63 ± 3.06bc | 40.12 ± 3.01b |
RIP(100 mg/kg) | 21.12 ± 1.43 | 26.82 ± 1.09 | 31.16 ± 1.24 | 33.73 ± 1.91 | 36.79 ± 1.57 | 37.81 ± 2.47c | 38.27 ± 2.98bc | 41.12 ± 2.16b |
The effect of the RIP on kidney, liver, thymus and spleen indexes in aging mice
Index (mg/g·bw) | Group | |||||
---|---|---|---|---|---|---|
Normal control | Model control | Positive control | RIP (25 mg/kg) | RIP (50 mg/kg) | RIP (100 mg/kg) | |
liver | 42.27 ± 2.72 | 37.93 ± 3.49d | 40.43 ± 2.03ac | 38.59 ± 3.14ac | 40.54 ± 3.40bc | 40.72 ± 2.48bc |
kidney | 15.59 ± 1.46 | 13.74 ± 1.69d | 14.04 ± 1.41ac | 13.97 ± 1.20c | 14.39 ± 1.88ac | 14.76 ± 1.49ac |
spleen | 2.45 ± 1.30 | 2.17 ± 0.50c | 2.25 ± 0.75a | 2.29 ± 0.67ac | 2.30 ± 1.01ac | 2.37 ± 0.58a |
thymus | 2.48 ± 0.64 | 2.02 ± 0.87c | 2.32 ± 0.50a | 2.28 ± 0.34ac | 2.35 ± 0.50a | 2.36 ± 0.34a |
Histopathological observation
The effect of RIP on histological changes in livers
The effects of RIP on histology changes in kidneys
The effect of the RIP on the activities of antioxidant enzymes in liver, kidney and brain in aging mice
Group | MDA | TAOC | CAT | SOD | GSH-Px |
---|---|---|---|---|---|
Normal control | 16.93 ± 0.63 | 8.09 ± 0.42 | 29.01 ± 1.79 | 208.63 ± 27.29 | 1094.56 ± 69.89 |
Model control | 24.87 ± 0.89d | 6.52 ± 0.48d | 14.00 ± 1.74d | 157.25 ± 34.62d | 784.50 ± 44.70d |
Positive control | 19.77 ± 0.58ad | 8.24 ± 0.62b | 27.78 ± 1.53b | 207.49 ± 32.40b | 1069.13 ± 24.48b |
RIP (25 mg/kg) | 24.58 ± 0.39d | 7.15 ± 0.63c | 16.32 ± 2.33ad | 165.53 ± 18.84ad | 874.43 ± 31.82ad |
RIP (50 mg/kg) | 23.32 ± 0.52ad | 7.65 ± 0.35a | 24.83 ± 2.13bc | 187.74 ± 13.60bd | 950.97 ± 19.70bc |
RIP (100 mg/kg) | 20.99 ± 0.62bd | 8.58 ± 0.53b | 26.49 ± 1.33bc | 227.47 ± 16.31bc | 1167.18 ± 82.58b |
Group | MDA | TAOC | CAT | SOD | GSH-Px |
---|---|---|---|---|---|
Normal control | 1.43 ± 0.10 | 1.22 ± 0.15 | 36.18 ± 0.30 | 171.39 ± 16.44 | 399.67 ± 16.92 |
Model control | 2.78 ± 0.09d | 0.96 ± 0.11c | 28.05 ± 3.28d | 100.89 ± 3.77d | 343.89 ± 19.44a |
Positive control | 1.34 ± 0.09b | 2.08 ± 0.33bd | 49.67 ± 2.24bd | 116.71 ± 8.18ad | 479.49 ± 23.56bd |
RIP (25 mg/kg) | 2.22 ± 0.14ad | 1.09 ± 0.14c | 35.51 ± 1.34b | 107.36 ± 10.86d | 345.65 ± 12.35c |
RIP (50 mg/kg) | 2.01 ± 0.10ab | 1.59 ± 0.17ac | 39.41 ± 1.97bc | 124.62 ± 11.53ad | 378.94 ± 15.05a |
RIP (100 mg/kg) | 1.80 ± 0.10ac | 1.78 ± 0.22bc | 42.57 ± 2.19bd | 136.92 ± 9.15bd | 414.85 ± 19.95dc |
Group | MDA | TAOC | CAT | SOD | GSH-Px |
---|---|---|---|---|---|
Normal control | 1.46 ± 0.24 | 1.69 ± 0.10 | 32.77 ± 2.44 | 102.35 ± 5.42 | 248.16 ± 8.14 |
Model control | 2.38 ± 0.26d | 0.88 ± 0.08d | 25.44 ± 1.78d | 74.33 ± 6.96d | 174.23 ± 7.92d |
Positive control | 1.17 ± 0.28bc | 1.61 ± 0.12b | 34.99 ± 0.92b | 87.27 ± 8.00ad | 259.39 ± 10.37bc |
RIP (25 mg/kg) | 1.84 ± 0.38ac | 1.28 ± 0.12ac | 26.36 ± 1.72d | 76.00 ± 9.04d | 186.41 ± 8.45ad |
RIP (50 mg/kg) | 1.67 ± 0.29a | 1.42 ± 0.16bc | 32.62 ± 1.89a | 92.10 ± 4.59bc | 212.57 ± 6.82ac |
RIP (100 mg/kg) | 1.30 ± 0.35a | 1.81 ± 0.28b | 43.92 ± 0.87bd | 112.48 ± 6.13bc | 236.78 ± 9.15bc |
Group | MDA | TAOC | CAT | SOD | GSH-Px |
---|---|---|---|---|---|
Normal control | 1.07 ± 0.07 | 1.01 ± 0.15 | 32.07 ± 2.02 | 258.22 ± 12.68 | 319.32 ± 8.94 |
Model control | 1.63 ± 0.27d | 0.64 ± 0.09d | 25.75 ± 2.23d | 162.78 ± 23.86d | 228.75 ± 6.95d |
Positive control | 0.89 ± 0.08bc | 1.27 ± 0.12bc | 49.67 ± 2.24bd | 250.47 ± 11.65b | 354.29 ± 11.28bc |
RIP (25 mg/kg) | 1.61 ± 0.07 | 0.66 ± 0.04 | 24.27 ± 2.53 | 164.91 ± 17.81 | 224.87 ± 12.62 |
RIP (50 mg/kg) | 1.58 ± 0.10 | 0.67 ± 0.08 | 25.33 ± 3.19 | 165.98 ± 15.02 | 227.56 ± 14.89 |
RIP (100 mg/kg) | 1.55 ± 0.10 | 0.71 ± 0.20 | 25.40 ± 2.56 | 165.47 ± 11.65 | 229.36 ± 15.63 |