Figure 1.
Structure of anthocyanin in blackcurrants. The chemical structure of each of the four anthocyanins (ACs) is as follows: D3R (X = OH, Y = glucosylrhamnose), D3G (X = OH, Y = glucose), C3R (X = H, Y = glucosylrhamnose) and C3G (X = H, Y = glucose).
Figure 1.
Structure of anthocyanin in blackcurrants. The chemical structure of each of the four anthocyanins (ACs) is as follows: D3R (X = OH, Y = glucosylrhamnose), D3G (X = OH, Y = glucose), C3R (X = H, Y = glucosylrhamnose) and C3G (X = H, Y = glucose).
Figure 2.
Schematic diagram of rhodopsin regeneration by anthocyanin.
Figure 2.
Schematic diagram of rhodopsin regeneration by anthocyanin.
Figure 3.
AC effects of the rhodopsin regeneration time course. (•) control; (◇) D3G; (◆) D3R; (△) C3G; (▲) C3R. Rhodopsin regeneration was fitted to an exponential function of 1-exp (–t/τ). Control, τ = 114 sec (R2 = 0.997); D3G, τ = 122 sec (R2 = 0.988); D3R, τ = 119 sec (R2 = 0.993); C3G, τ = 94 sec (R2 = 0.994) and C3R, τ = 89 sec (R2 = 0.994). Values are the mean of nine experiments, and vertical bars represent SEM.
Figure 3.
AC effects of the rhodopsin regeneration time course. (•) control; (◇) D3G; (◆) D3R; (△) C3G; (▲) C3R. Rhodopsin regeneration was fitted to an exponential function of 1-exp (–t/τ). Control, τ = 114 sec (R2 = 0.997); D3G, τ = 122 sec (R2 = 0.988); D3R, τ = 119 sec (R2 = 0.993); C3G, τ = 94 sec (R2 = 0.994) and C3R, τ = 89 sec (R2 = 0.994). Values are the mean of nine experiments, and vertical bars represent SEM.
Figure 4.
Effect of C3R on the initial rate of rhodopsin regeneration. Initial velocity of rhodopsin regeneration was determined for 40 s in the C3R (○) and control (●) at each 11-cis-retinal concentration. The data points were fitted with eq 2 to calculate rate constants in the regeneration reaction (n = 6, mean ± SE).
Figure 4.
Effect of C3R on the initial rate of rhodopsin regeneration. Initial velocity of rhodopsin regeneration was determined for 40 s in the C3R (○) and control (●) at each 11-cis-retinal concentration. The data points were fitted with eq 2 to calculate rate constants in the regeneration reaction (n = 6, mean ± SE).
Figure 5.
Ciliary muscle relaxation by D3R against ET-1induced contraction. 10 nM ET-1 produced a phasic contraction, followed by a tonic contraction. When the tonic contraction reached a steady level, 30 µM D3R or H2O (vehicle) was added and the resulting relaxation was monitored for 60 min. SNP (sodium nitroprusside) is a positive control of relaxation as an NO donor. W/O is a washout of all reagents and solution.
Figure 5.
Ciliary muscle relaxation by D3R against ET-1induced contraction. 10 nM ET-1 produced a phasic contraction, followed by a tonic contraction. When the tonic contraction reached a steady level, 30 µM D3R or H2O (vehicle) was added and the resulting relaxation was monitored for 60 min. SNP (sodium nitroprusside) is a positive control of relaxation as an NO donor. W/O is a washout of all reagents and solution.
Figure 6.
Summary of data obtained from relaxation during a 60 minute treatment of four flavonoids compared to the control. * p < 0.05, ** p < 0.01; significantly different compared to the control. The data represent the force generated with ET-1 (10 nM) set to 100%. n = 6, mean ± SEM.
Figure 6.
Summary of data obtained from relaxation during a 60 minute treatment of four flavonoids compared to the control. * p < 0.05, ** p < 0.01; significantly different compared to the control. The data represent the force generated with ET-1 (10 nM) set to 100%. n = 6, mean ± SEM.
Figure 7.
Schematic diagram of relaxation induced by D3R.
Figure 7.
Schematic diagram of relaxation induced by D3R.
Figure 8.
Photograph of a chick wearing a negative lens (–8D).
Figure 8.
Photograph of a chick wearing a negative lens (–8D).
Figure 9.
The blackcurrant anthocyanin (BCA) effect on enlargement of each eye component dimensions after application of a negative lens (–8D) in chicks. n = 12, mean ± SEM. paired t-test (intra-group) or Dunnett's multiple comparison test (inter-group). * p < 0.05, and ** p < 0.01; vs. control.
Figure 9.
The blackcurrant anthocyanin (BCA) effect on enlargement of each eye component dimensions after application of a negative lens (–8D) in chicks. n = 12, mean ± SEM. paired t-test (intra-group) or Dunnett's multiple comparison test (inter-group). * p < 0.05, and ** p < 0.01; vs. control.
Figure 10.
Typical dark adaptation curve before (continuous line) and after (dotted line) BCA intake.
Figure 10.
Typical dark adaptation curve before (continuous line) and after (dotted line) BCA intake.
Table 1.
The ocular tissue distribution of AC at 1 h post-intraperitoneal administration in rats and at 30 min post-intravenous administration in rabbits. Mean ± SE, n = 5 in rats, and n = 3 in rabbits.
Table 1.
The ocular tissue distribution of AC at 1 h post-intraperitoneal administration in rats and at 30 min post-intravenous administration in rabbits. Mean ± SE, n = 5 in rats, and n = 3 in rabbits.
Ocular Tissues or Body Fluid | Intraperitoneal Administration in Rat | Intravenously Administration in Rabb |
---|
AC of Tissue (μg/g Tissue) | Distribution Ratio (%) | AC of Tissue (μg/g Tissue) | Distribution Ratio (%) |
---|
Aqueous humor | 6.72 | 0.88 | 1.19 ± 0.21 | 10.54 |
Cornea | 20.62 | 3.67 | 0.55 ± 0.05 | 4.89 |
Sclera | 245.04 | 89.09 | 3.02 ± 0.09 | 26.73 |
Choroid | 3.00 ± 0.06 | 26.57 |
Ciliary body | 12.93 | 1.39 | 2.04 ± 0.28 | 18.07 |
Iris | 1.11 ± 0.08 | 9.81 |
Retina | 6.89 | 4.76 | 0.27 ± 0.02 | 2.41 |
Vitreous | 0.60 | 0.14 | 0.11 ± 0.02 | 0.98 |
Lens | 0.36 | 0.06 | 0.00 ± 0.00 | 0.00 |
Plasma * | 2.30 ± 0.76 | | 12.42 ± 1.25 | |
Table 2.
Kinetic parameters of rhodopsin regeneration.
Table 2.
Kinetic parameters of rhodopsin regeneration.
| Control * | +C3R * |
---|
k2 (s−1) | 1.0 × 10−2 | 1.1 × 10−2 |
km (M) | 2.6 × 10−5 | 1.1 × 10−5 |
Table 3.
Comparison of contraction of the ciliary muscle by ET-1 (10 nM) after treatment with various reagents.
Table 3.
Comparison of contraction of the ciliary muscle by ET-1 (10 nM) after treatment with various reagents.
Treatment | Concentration | n | Contraction (%) (Average ± SE) |
---|
control | | 21 | 54.9 ± 3.3 a |
D3R | 10 mM | 21 | 42.2 ± 3.2 |
D3R + NOARG | 10 mM + 10 mM | 12 | 63.7 ± 7.1 a |
D3R + NOARG+l-Arg | 10 mM + 10 mM + 10 mM | 12 | 42.7 ± 4.2 |
Carboxy-PTIO | 30 mM | 10 | 54.4 ± 2.1 |
D3R + Carboxy-PTIO | 10 mM + 30 mM | 10 | 55.2 ± 3.2 a |
ODQ | 10 mM | 11 | 56.1 ± 5.4 |
D3R + ODQ | 10 mM + 10 mM | 11 | 58.5 ± 5.5 a |
BQ788 | 10 μM | 12 | 81.0 ± 8.2 |
D3R + BQ788 | 10 mM + 10 μM | 12 | 73.4 ± 9.6 a |
propranolol | 10 mM | 9 | 61.3 ± 3.6 |
D3R + propranolol | 10 mM + 10 mM | 8 | 48.7 ± 3.4 b |
iberiotoxin | 10 μM | 12 | 64.6 ± 3.0 |
D3R + iberiotoxin | 10 mM + 10 μM | 12 | 45.7 ± 4.8 c |
indomethacin | 10 mM | 12 | 70.3 ± 5.3 |
D3R + indomethacin | 10 mM + 10 mM | 12 | 52.4 ± 5.2 d |
Table 4.
Comparison between ET-1 induced ciliary muscle contraction ratio and phosphorylated MLC ratio.
Table 4.
Comparison between ET-1 induced ciliary muscle contraction ratio and phosphorylated MLC ratio.
Treatment | Concentration | Contraction (%) (Average ± SE) | Phosphorylated-Ratio (%) (Average ± SE) |
---|
control | | 54.9 ± 3.3 a | 47.2 ± 13.1 a |
D3R | 10 mM | 42.2 ± 3.2 | 36.0 ± 13.6 |
D3R + NOARG | 10 mM + 10 mM | 63.7 ± 7.1 a | 50.1 ± 4.2 a |
D3R + NOARG + l-Arg | 10 mM + 10 mM+ 10 mM | 42.7 ± 4.2 b | 38.2 ± 3.7 c |
Table 5.
Dark adaptation threshold changes before and after intake of the test drink.
Table 5.
Dark adaptation threshold changes before and after intake of the test drink.
Dose of BCA | Dark Adaptation Value, Mean ± SD of Log Asb; (p value) (1) |
---|
mg/subject | Before Intake | After Intake | Change | p value (2) |
---|
0 (placebo) | 2.056 ± 0.209 (1.000) | 2.018 ± 0.218 (1.000) | −0.038 ± 0.106 (1.000) | 0.244 |
12.5 | 2.026 ± 0.147 (0.457) | 2.004 ± 0.195 (0.457) | −0.023 ± 0.138 (0.733) | 0.583 |
25 | 2.016 ± 0.170 (0.234) | 1.980 ± 0.197 (0.264) | −0.037 ± 0.112 (0.983) | 0.28 |
50 | 2.038 ± 0.186 (0.686) | 1.923 ± 0.167 (0.014) | −0.115 ± 0.131 (0.171) | 0.011 |
Table 6.
Refraction values in dominant eye, flicker values, assessment of subjective asthenopia and changes in transient refractive alteration measured before and after a visual task in subjects orally administered BCA or placebo 1.
Table 6.
Refraction values in dominant eye, flicker values, assessment of subjective asthenopia and changes in transient refractive alteration measured before and after a visual task in subjects orally administered BCA or placebo 1.
Item & Statement | BCA | Placebo |
---|
Before | After 2 | Change 2 | Before | After 2 | Change 2 |
---|
Refraction values | −0.432 ± 0.602 | −0.402 ± 0.643 a | −0.030 ± 0.252 (a),c | −0.384 ± 0.536 | −0.503 ± 0.579 b | 0.119 ± 0.278 (b), (c) |
Flicker value, Hz | 34.95 ± 3.16 | 34.39 ± 3.51 | 0.56 ± 1.15 | 34.72 ± 2.99 | 34.13 ± 2.90 | 0.59 ± 1.22 |
Asthenopia symptoms (VAS mm) | | | | | | |
head & neck | 12.70 ± 13.45 | 40.08 ± 24.86 | 27.38 ± 18.39 | 8.34 ± 11.87 | 44.09 ± 26.09 | 35.75 ± 24.96 |
arm | 10.21 ± 17.82 | 36.15 ± 25.86 | 25.94 ± 29.61 | 4.32 ± 6.27 | 41.76 ± 29.33 | 37.44 ± 28.43 |
eye | 14.72 ± 15.55 | 47.31 ± 24.72 | 32.59 ± 18.94 (a) | 14.59 ± 17.98 | 56.72 ± 25.24 | 42.14 ± 19.52 (b) |
shoulder | 15.12 ± 15.65 | 49.66 ± 27.97 | 34.54 ± 25.75 | 10.95 ± 17.42 | 54.31 ± 29.31 | 43.36 ± 30.91 |
low back | 10.63 ±15.92 | 29.79 ± 27.15 a | 19.16 ± 22.74 (a) | 7.35 ± 9.25 | 42.83 ± 33.55 b | 35.48 ± 30.87 (b) |
Table 7.
Retinal blood flow of the superior temporal rim/retina and inferior temporal/retina after consumption of anthocyanins for 6 months in the left eye (LE) and right eye (RE).
Table 7.
Retinal blood flow of the superior temporal rim/retina and inferior temporal/retina after consumption of anthocyanins for 6 months in the left eye (LE) and right eye (RE).
Treatment | Before (Average ± SE) | After 6 Month (Average ± SE) |
---|
Sup. temp. rim (RE) | 507.7 ± 174.3 | 638.6 a ± 191.2 |
Inf. temp. rim (RE) | 393.6 ± 138.0 | 582.2 b ± 177.8 |
Sup. temp. retina (RE) | 457.6 ± 140.6 | 595.1 b ± 171.5 |
Inf. Temp. retina (RE) | 377.0 ± 80.5 | 519.1 b ± 130.0 |
Sup. temp. rim (LE) | 442.4 ± 214.3 | 662.4 b ± 185.3 |
Inf. temp. rim (LE) | 466.5 ± 216.3 | 653.7 b ± 260.9 |
Sup. temp. retina (LE) | 375.0 ± 75.9 | 442.2 b ± 80.1 |
inf. temp. rim (LE) | 444.9 ± 100.9 | 546.9 a ± 185.8 |