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Session CS15.3 Scientific Foundation of CM. Neuroprotection of Pre-germinated Brown Rice in Rat Cortical Neuron. Hsin-Hsueh Lee * , Chi-Ting Lin, Chien-Lung Hung and Ling-Ling Yang # Department of Pharmacognosy, College of Pharmacy, Taipei Medical University, Taipei, Taiwan 24 th , Nov. 2006.
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Session CS15.3 Scientific Foundation of CM Neuroprotection of Pre-germinated Brown Rice in Rat Cortical Neuron Hsin-Hsueh Lee*, Chi-Ting Lin, Chien-Lung Hung and Ling-Ling Yang# Department of Pharmacognosy, College of Pharmacy, Taipei Medical University, Taipei, Taiwan 24th, Nov. 2006 2006 World Congress on Chinese Medicine in HK
PGBR • pre-germinated brown rice (PGBR) which brown rice is soaked in water to induce slight germination. BR GBR low temp. dry PGBR 22h after germinated
Pharmacology of PGBR Hypocholesterolemic action of pre-germinated brown rice in hepatoma-bearing rats. (Miura D, et al.,2006 Life Sci.) Postprandial blood glucose and insulin responses to pre-germinated brown rice in healthy subjects. (ItoY, et al., 2005 J Med Invest.) Insoluble fiber is a major constituent responsible for lowering the post-prandial blood glucose concentration in the pre-germinated brown rice.(Seki T, et al., 2005 Biol Pharm Bull.)
Pharmacology of PGBR Effects of pre-germinated brown rice on beta-amyloid protein-induced learning and memory deficits in mice. (Mayima T, et al., 2004 Biol Pharm Bull.) Effects of germinated brown rice extracts with enhanced levels of GABA on cancer cell proliferation and apoptosis.(Oh CH and Oh SH, 2004 J Med Food.)
Materials • Brown rice (germinated rate > 95%) was provided by Asia Rice Co. Taiwan • Wistar Rat was purchased from the animal center of National Taiwan University, Taiwan.
Methods • GABA amount detection: UV, HPLC • Anti-radical ability: DPPH, Fe2+ chelating, etc. • Neuroprotective investigation: MTT & LDH
Protein analysis in SDS page Marker RiceBR PGBR Marker RiceBR PGBR 40 μg 20 μg
AABA GABA GABA Quantification by HPLC mAU Min HPLC (Shimadzu 10Avp) Column:LiChroCart LiChrospher 100 RP-18e, 250×4 mm I.D. (5 μm) Detector: 532nm Mobile Phase: 60 % acetonitrile in water Flow Rate:1ml/min
Anti-radical Effects of PGBR PGBR IC50 Value μg/ml Fe2+ IC50= 38±1.75 .OH-IC50= 103±6.24 DPPH IC50= 50±2.24 Table 1. The IC50 value of each anti-radical investigation of PGBR. N=4
A.A 6-HODA Glutamate Primary Cultured Cortical Neuron mature neuron Seeding MTT and LDH assay 0 DIV 11~12 DIV Preparation of Wistar Rat 1 h 24 h Treat the cells with PGBRE and inducers Sacrificed the rats and cultured the cortical neurons 12DIV
Effect of Rice in Cultured Neuron *, p<0.05; **, p<0.01 compared with CTL
The Neurotoxic Insults • 6-HODA: parkinsonism • Glutamate: excitatory amino acid • Arachidonic acid: inflammatory cascade
Effect of PGBRE on 6-HODA-induced Neurotoxicity μg/ml μg/ml *, p<0.05; **, p<0.01 compared with CTL
Effect of PGBRE on Arachidonic Acid-induced Neurotoxicity μg/ml μg/ml *, p<0.05; **, p<0.01 compared with CTL; +, p<0.05; ++p<0.01 compared with A.A
Effect of PGBRE on glutamate-induced Neurotoxicity μg/ml μg/ml *, p<0.05; **, p<0.01 compared with CTL; +, p<0.05; ++p<0.01 compared with Glu. NMDA as positive control
Summary • PGBR had the most GABA content in brown rice. • PGBR exerted the potential anti-radical ability. • PGBR showed significantly preventive effect against neurotoxic insults in neuron.
Deduced Possibility of PGBR on Neuroprotection PGBR Fe2+、DPPH GABA ROS Glutamate Arachidonic acid Neuroprotection
Conclusion Pre-germinated brown rice could significantly decreased glutamate and arachidonic acid- induced neurotoxicity. PGBR could be benefit for brain injury and neurodegeneration.
Acknowledgement This work was supported by Dr. Chi-Chin Huang Foundation in the USA (4152Z23-B30-3) and we also thanks to topnotch stroke research center grant, Taiwan Ministry of Education.
Session CS15.3 Scientific Foundation of CM We are looking forward to your comments and suggestions! 2006 World Congress on Chinese Medicine in HK