Abstract��OBJECTIVE To observe the change of EPO in brain of aging rat induced by D-galactose(D-gal) and the EPO-based antiaging of the water extract of Radix Rehmanniae Preparata(WERRP). METHODSD-gal-treated groups were received subcutaneous injection of D-gal at dose of 50, 150 and 250 mg��kg-1 daily for 8 weeks to imitate an aging model that was induced by oxidative stress. After the detection of EPO in hippocampus, the 150 mg��kg-1D-gal group was chose as the aging model. In addition the WERRP-treated group and vehicle group was set. The WERRP-treated group was given WERRP oral gavage at a dose of 4 g��kg-1 daily starting from the 5th week. Morris water maze (MWM) test was used to assess the spatial learning and memory. SOD, MDA and ��-galactosidase in brain were examined by Assay Kits. Finally, EPO, EPOR, and HIF-2�� in hippocampus were determined by immunohistochemistry and Western blot. RESULTSD-gal-treated group showed significant longer latency to platform and less times of cross the platform(P<0.05) in MWM. After treated with D-gal, SOD was drastically decreased and MDA and ��-galactosidase were remarkably increased in brains compared with vehicle (0.9% saline)-treated rats (P<0.01). In addition, the expression of EPO,EPOR and HIF-2�� were significantly decreased in the brains of D-gal-treated rats compared with vehicle-treated rats. Meanwhile, there was a negative correlation between EPOR and MDA in content. Interestingly, WERRP-treated rats showed significant improvement of spatial learning and memory, decrease of oxidative stress and enhancement of EPO/EPOR in the brain compared with 150 mg��kg-1D-gal-treated rat(P<0.05). CONCLUSION The aging rats induced by D-gal show a significant decline of EPO in brain which indicate the decrease of brain EPO in aging is related to the increase of oxidative stress. That WERRP reverses the decline of the EPO expression in aging model may be the underlying mechanism of the role of anti-aging of WERRP.
����, ��Ȼ, ������, ͯ��, ������. D-��������˥�ϴ������ڴٺ�ϸ�������ر���µ�����ػƵ���ת����[J]. �й�ҩѧ��־, 2016, 51(18): 1562-1568.
LI Xu, HE Ran, WANG Hong-yu, TONG Jie, XU Xiao-yu. The Decline of EPO in Brain of Aging Rats Induced by D-galactose Effect and the Reverse of the Water Extract of Radix Rehmanniae Preparata. Chinese Pharmaceutical Journal, 2016, 51(18): 1562-1568.
XU X, XU D H, DAI H B. Effect of erythropoietin on neuron death induced by hypoxia [J]. Chin Pharm J(�й�ҩѧ��־), 2005,16(40):1223-1226.
KADOTA T, SHINGO T, YASUHARA T, et al. Continuous intraventricular infusion of erythropoietin exerts neuroprotective/rescue effects upon Parkinson's disease model of rats with enhanced neurogenesis[J]. Brain Res, 2009, 12(54): 120-127.
NADAM J, NAVARRO F, SANCHEZ P, et al. Erythropoietin protects hippocampal neurons following status epilepticus[J]. Neurobiol Disease, 2007, 25(2): 412-426.
LEE S T, CHU K, PARK J E, et al. Erythropoietin improves memory function with reducing endothelial dysfunction and amyloid-beta burden in Alzheimer's disease models[J]. J Neurochemistry, 2012, 120(1): 115-124.
JIA Z, XUE R, MA S, et al. Erythropoietin attenuates the memory deficits in aging rats by rescuing the oxidative stress and inflammation and promoting BDNF releasing[J]. Mol Neurobiol, 2015, DOI: 10. 1007/s12305-015-9438-1.
YOON H C, SOO I K, KYEUNG M J, et al. Age-related changes in erythropoietin immunoreactivity in the cerebral cortex and hippocampus of rats[J]. Brain Res, 2004, 1018(1):141-146.
JUUL S E, HARCUM J, LI Y, et al. Erythropoietin is present in the cerebrospinal fluid of neonates [J]. J Pediatrics, 1997, 130(3):428-430.
HAN C H, LIN Y F, LIN Y S, et al. Effects of yam tuber protein, dioscorin, on attenuating oxidative status and learning dysfunction in D-galactose-induced BALB/c mice[J]. Food Chem Toxicol, 2014, 65(1):356-363.
LI P F, MIAO M S. Analysis on modern research and application status of Prepared Radix Rehmanniae[J]. China J Chin Med(��ҽѧ��), 2014,29( 2) :252-254.
ZHU H F,WANG D,XU X Y, et al. Effects of rehmannia root decoction serum on cell proliferation and EPO expression in cultured human umbilicalvein endotheilial cells[J]. China J Chin Mater Med (�й���ҩ��־), 2008, 33(13):1579-1582.
XU C, WANG L, ZUO P, et al. D-Galactose-caused life shortening in Drosophila melanogaster and Musca domestica is associated with oxidative stress[J]. Biogerontol, 2004, 5(5):317-325.
CUI Y, YAN Z H, HOU S L, et al. Effect of Shu di-huang on the transmitter and receptor of amino acid in brain and learning and memory of dementia model[J]. China J Chin Mater Med (�й���ҩ��־),2003, 28(9):862-866.
RATCLIFFE P J. HIF-1 and HIF-2: working alone or together in hypoxia? [J]. J Clin Investigation, 2007, 117(4):862-865.
HWANG A B, LEE S J. Regulation of life span by mitochondrial respiration: the HIF-1 and ROS connection[J]. Aging, 2011, 3(3):304-310.