H Matsumoto [Mode de compatibilité]

New Finding of the Visual ImprovementEffect of Blackcurrant Anthocyanins IntakeHitoshi MatsumotoFood and Health R&D Laboratories,Meiji Seika Kaisha, Ltd.

TThe Comparison of Bioavailabilitybetween Blackcurrant and BilberrySample: 1)Blackcurrant Anthocyanin (BCA:Currantex20S(JTB))2)Bilberry Anthocyanin(BBA:Bilberon25(Tokiwa, Japan))Dose: Anthocyanin 400mg/kg/body weightIngestion: Single oral ingestionEvaluation:Ⅰ)Anthocyanin concentration in plasma and eye ballafter 0.5 and 1 hour administration.Ⅱ)Urinary excretion amount of anthocyaninuntil 24 hour after administration

Anthocyanin Component in Blackcurrant and BilberryOnly BlackcurrantBlackcurrant4componentBilberry15componentXOH+HOOYO ZOHStructure of AnthocyaninAC content% X Y ZDel-3-RutDel-3-GlcCya-3-RutCya-3-Glc47.013.734.74.6Del-3-Gal 12.79Del-3-Glc 12.20Del-3-Ara 10.76Cya-3-Gal 11.02Cya-3-Glc 10.63Cya-3-Ara 8.09Pet-3-Gal 3.98Pet-3-Glc 7.57Pet-3-Ara 2.48Peo-3-Gal 1.04Peo-3-Glc 5.15Peo-3-Ara 0.65Mal-3-Gal 4.37Mal-3-Glc 7.89Mal-3-Ara 1.37OHOHHOHOHHOHOCH 3 HOCH 3Glc-RhamGlcGlc-RhamGlcGalGlcAraGalGlcAraGalGlcAraGalGlcAraGalGlcAra

AC Conc (ng/mL)Excretion rate (%)AC concentration in rat plasma500040003000200010000TThe Comparison of Bioavailabilitybetween Blackcurrant and Bilberryp

Bioavailability of BlackcurrantAnthocyanin is higher than bilberryanthocyaninBlackcurrant Anthocyanin is the bestfor eye supplement

Visual Improvement Effect ofBlackcurrant Anthocyanin(1) Asthenopia (Eye fatigue) StudyNakaishi, et. al., Altern.Med.Rev., 5(6), 553-562, 2000 (Clinical Study)Matsumoto, et al., Journal of the eye., 23(1), 129-133, 2006 (Clinical Study)Iida, et.al., Journal of the eye., 25(1), 114-118, 2008 (Clinical Study)(2) Improvement of Dark Adaptation StudyH. Nakaishi, et. al Altern.Med.Rev., 5(6), 553-562, 2000 (Clinical Study)(3) Anti-Glaucoma StudyI. Ohguro, et. al Hirosaki Med. J., 59, 23-32, 2007 (Clinical Study)

(1) Asthenopia (Eye Fatigue) Study(Methods)Subjects; 21 healthy volunteers (age 20-25, mean 20.9)Design; Double-blind, randomized, placebo-controlled, crossover studyDose; 50mg of anthocyanin / subject, taking 2 hr before taskVisual task; Simple calculation task on 2 hours without pauseEvaluation: Degradation of refraction, Visual Analog Scale questionnaireintake measurement measurement10min2hrrest10min2hrVDT task10minPhotoVisual Analog Scale Questionnaireno fatiguestrong fatiguePhotoVisual taskTThe subject’s response to each statement was definedas the distance from the left end of the line to the mark.Auto-refractometerH. Nakaishi, et. al Altern.Med.Rev., 5(6), 553-562, 2000

Results% 80604020p =0.037p =0.025BCAplaceboDiopter(D)-0.35-0.40-0.45-0.50p=0.598p=0.064p=0.006PlaceboBlackcurrantanthocyanin0Head &neckarm eye shoulder low back-0.55Before taskAfter taskH. Nakaishi, et. al Altern.Med.Rev., 5(6), 553-562, 2000

(2) Improvement of Dark Adaptation StudyPhoto摂 取 前 測 定摂 取 後 測 定2 分 210min分 30min分120min2min 分10min 分 30min分2minpre-intake measurement2 時 間Post-intake measurementGoldmann Adaptometer暗 順 応Dark adaptation摂 取intake暗 順 応Dark adaptationDesign: double-blind, placebo-controlled crossover studyWash out is 10daysSample: 50, 25, 12.5, 0 mg Blackcurrant anthocyaninfilled in capsuleSubjects: 12 healthy volunteer, Ave 33.3 years old.Evaluation: Measurement of dark adaptation curve.Luminance at 30min is Dark adaptation threshold.Dark Adaptation Curve H. Nakaishi, et. al Altern.Med.Rev., 5(6), 553-562, 2000

Results[[log(asb)]]threshold-4.1-4.2-4.3-4.4-4.5-4.6-4.7p =0.011pre postplaceboBCA 50mgChange of threshold [[log(asb)]]0.10.0-0.1-0.2-0.3p =0.0110 10 20 30 40 50dose [[mg AC]]Ave ±SD, n=12H. Nakaishi, et. al Altern.Med.Rev., 5(6), 553-562, 2000

Anti-Glaucoma StudySubject: Normal tension glaucoma patient inHirosaki University glaucoma clinicGender: 6 Male, 21 FemaleAge: 51~80 years old(66.7 ± 6.9)Sample: Blackcurrant Anthocyanin (50mg/day)6month oral intakeEvaluation1Blood Flow at the neuroretinal rimof the optic nerve head and peripapillary retina2Intraocular pressure3Visual fieldGlaucoma eyePhotoI. Ohguro, et. al Hirosaki Med. J., 59, 23-32, 2007

ResultsBeforeAfter 6monthIOP (RE)(mmHg) 13.9 ± 1.813.5 ± 1.6IOP(LE)(mmHg) 13.6± 2.113.4 ± 1.9MD(RE) (dB) -3.28 ± 4.65 -3.23 ± 4.49MD(LE) (dB) -4.51 ± 4.34 -4.32 ± 4.29MD(RE)(dB)1055MD(LE)(dB)01 201 2-5-5-10-10-15-15-20-20

Blood Flow at the Neuroretinal Rim of theOptic Nerve Head and Peripapillary RetinaSup. temp. rim (RE)Inf. temp. rim (RE)Sup. temp. retina (RE)Inf. TTemp. retina (RE)Sup. temp. rim (LE)Inf. temp. rim (LE)Sup. temp. retina (LE)inf. temp. rim (LE)Before507.7 ±174.3393.6 ±138.0457.6 ±140.6377.0 ±80.5442.4 ±214.3466.5 ±216.3375.0 ±75.9444.9 ±100.9After 6 month638.6 * ±191.2582.2 ** ±177.8595.1 ** ±171.5519.1 *** ±130.0662.4 *** ±185.3653.7 *** ±260.9442.2 ** ±80.1546.9 * ±185.8*p < 0.05, ** p < 0.01, *** p< 0.0005

Preventive Effect of Black Currant on NegativeLens-Induced Myopia in a Chick ModelPhoto

Background• Myopia is a common visual disorder/disease ofyoungsters, particularly in East Asia.• In Taiwan, the prevalence rate is more than 85% atthe age of 18.• There is a very high correlation between thechange in axial length and progression of myopiain school children (6 to 12 years old).• Myopia in chicks, induced by wearing a negativelens, is considered to be a good model for humanmyopia during school-age years.

Mechanism of Axial Length Elongation by Near-WorkCorneaRetinaHyperopic defocusNear workSchool children possess a largeamplitude of accommodation andsustained accommodation resultsin an elongation of axial length.FAxial length elongatesMyopia develops

Aim・To establish the negative lens-inducedchick myopia model.・To evaluate the effects of orallyadministered black currant (BC) extracton the enlargement of globe componentdimensions induced by wearing anegative lens in chicks.

Theory of Negative Lens-Induced MyopiaObjectHyperopicdefocusNormal eyeChick +Negative lens(simulates a near-work)Near workAxial length elongates toChick + Negative lenscompensate for the defocus

Methods of Chick Myopia ModelDay 1 Day 2 Day 3 Day 4Lens wearing●●●Measurement of axial lengthExperimental animal : 8-day-old chickLens placement:Negative (-8D) lens was placed infront of the right eye.Left (No lens) eye acted as a control.Measurement of axial length :A-scan ultrasound instrument under anesthesia

Apparatus of Chick Myopia ModelNegative (-8D) lens8-day-old chickAxial length was measured by A-scan as the ultrasound tracing.CorneaRetinaChick under anesthesiaAxial length

Results of Chick Myopia Model(Axial Length)Axial length (mm)9.209.109.008.908.808.708.60P=0.0110Day 2 Day 3 Day 4: Left eye (fellow eye): Right eye (negative lens)Each column shows the mean ± SEM of 4 chicks.P-values : paired t-test

Results of Chick Myopia Model(Ocular Length)Ocular length (mm)9.409.309.209.109.008.908.808.708.608.50P=0.011Day 2 Day 3 Day 4: Left eye (fellow eye): Right eye (negative lens)Each column shows the mean ± SEM of 4 chicks.P-values : paired t-test

Methods for Evaluating the Effects of BC ExtractDay 1 Day 2 Day 3 Day 4▲ ▲ ▲ ●P.O.Lens placementP.O.Experimental animal : 8-day-old chickP.O.Measurement of globecomponent dimensionsCompounds: 200, 400 mg of BC extract / kg, p.o. for 3 daysor distilled water (Control)Evaluation: Enlargement of globe components (mm) =lens-treated right eyes (mm) – untreated fellow eyes (mm)

Blackcurrant Anthocyanin (BCA)Black currant extract25% of anthocyanin contentCurrantex 20SJust the Berry Ltd.New ZealandD3RD3GC3GC3RAnthocyanin(AC) structure of BCAHOOHCl -+OOHOHXOGlycosideHPLC profiles of BCAAC contentDelphinidin-3-glucoside (D3G), X=OH, 12.3%Delphinidin-3-rutinoside (D3R), X=OH, 38.0%Cyanidin-3-glucoside (C3G), X=H, 7.3%Cyanidin-3-rutinoside (C3R), X=H, 42.4%

Effects of BC Extract on Enlargement of Globe ComponentDimensionsThe difference between lens-treated anduntreated eye (mm)0.700.600.500.400.300.200.100.00-0.10-0.20**Control 200 400BC extract (mg/kg, p.o.)*: anterior chamber depth: lens thickness: vitreous chamber depth: axial length: ocular length*****Each column shows the mean ± SEM of 16 chicks. P-values were calculated usinga paired t-test (intra-group) and Dunnett's multiple comparison test (inter-group)*P

Methods for Evaluating the Effects of BC AnthocyaninsI.V.Day 1 Day 2 Day 3 Day 4▲ ▲ ▲ ●Lens placementI.V.Experimental animal : 8-day-old chickI.V.Evaluation: Enlargement of globe components (mm) =Measurement of globecomponent dimensionsCompounds: D3G 1.86 µg/kg, D3R 6.72 µg/kg, C3G 1.28 µg/kgand 7.84 µg/kg of C3R were intravenously administeredto chicks in the wing vein under anesthesia for 3 days.lens-treated right eyes (mm) – untreated fellow eyes (mm)

Effects of BC Anthocyanins on Enlargement of Globe ComponentDimensionsThe difference between lens-treated anduntreated eye (mm)0.600.500.400.300.200.100.00-0.10-0.20Control (12) BCAs (11)** *: anterior chamber depth: lens thickness: vitreous chamber depth: axial lengthP-values were calculated using a paired t-test (intra-group) and Student's t-test(inter-group). *P

Conclusions• BC extract and Anthocyanins could inhibitenlargement of globe componentdimensions in a negative lens-induced chickmyopia model.• These findings suggest that orallyadministeredBCAs may suppress orprevent myopia progression in schoolchildren.

Thank you for your attention.These results will be also reported atICP 2010 in Montpellier.http://www1.montpellier.inra.fr/polyphenols2010/