Neuroprotection and spatial memory enhancement of four herbal mixture extract in HT22 hippocampal cells and a mouse model of focal cerebral ischemia

BMC Complementary and Alternative Medicine

Research Article

Cheol Min Kim¹  Young Whan Choi²  Yu Ri Kim³  Ha Neui Kim³  Byung Tae Choi⁴  Sung Min Ahn⁵  Jin Ung Baek⁶  Hwa Kyoung Shin⁷  Jae Won Lee⁸ 

[1] Department of Biochemistry, College of Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;Department of Horticultural Bioscience, College of Natural Resource and Life Science, Pusan National University, 626-706, Miryang, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;Department of Korean Medical Science, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Department of Korean Medical Science, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Division of Meridian and Structural Medicine, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;Department of Korean Medical Science, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;Division of Humanities and Social Medicine, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Division of Meridian and Structural Medicine, School of Korean Medicine, Pusan National University, 626-870, Yangsan, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;Division of Pharmacy, College of Pharmacy, Pusan National University, 609-735, Busan, Republic of Korea;Research Center for Anti-aging Technology Development, Pusan National University, 609-735, Busan, Republic of Korea;

关键词: Hippocampal cell;    Neuroprotection;    Memory;    Polygonum multiflorum;   

DOI  :  10.1186/s12906-015-0741-1

 received in 2014-10-30, accepted in 2015-06-23,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundFour traditional Korean medicinal herbs which act in retarding the aging process, Polygonum multiflorum Thunb., Rehmannia glutinosa (Gaertn) Libosch., Polygala tenuifolia Willd., and Acorus gramineus Soland., were prepared by systematic investigation of Dongeuibogam (Treasured Mirror of Eastern Medicine), published in the early 17th century in Korea. This study was performed to evaluate beneficial effects of four herbal mixture extract (PMC-12) on hippocampal neuron and spatial memory.MethodsHigh performance liquid chromatography (HPLC) analysis was performed for standardization of PMC-12. Cell viability, lactate dehydrogenase, flow cytometry, reactive oxygen species (ROS), and Western blot assays were performed in HT22 hippocampal cells and immunohistochemistry and behavioral tests were performed in a mouse model of focal cerebral ischemia in order to observe alterations of hippocampal cell survival and subsequent memory function.ResultsIn the HPLC analysis, PMC-12 was standardized to contain 3.09 % 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside, 0.35 % 3′,6-disinapoyl sucrose, and 0.79 % catalpol. In HT22 cells, pretreatment with PMC-12 resulted in significantly reduced glutamate-induced apoptotic cell death. Pretreatment with PMC-12 also resulted in suppression of ROS accumulation in connection with cellular Ca2+ level after exposure to glutamate. Expression levels of phosphorylated p38 mitogen-activated protein kinases (MAPK) and dephosphorylated phosphatidylinositol-3 kinase (PI3K) by glutamate exposure were recovered by pretreatment with either PMC-12 or anti-oxidant N-acetyl-L-cysteine (NAC). Expression levels of mature brain-derived neurotrophic factor (BDNF) and phosphorylated cAMP response element binding protein (CREB) were significantly enhanced by treatment with either PMC-12 or NAC. Combination treatment with PMC-12, NAC, and intracellular Ca2+ inhibitor BAPTA showed similar expression levels. In a mouse model of focal cerebral ischemia, we observed higher expression of mature BDNF and phosphorylation of CREB in the hippocampus and further confirmed improved spatial memory by treatment with PMC-12.ConclusionsOur results suggest that PMC-12 mainly exerted protective effects on hippocampal neurons through suppression of Ca2+-related ROS accumulation and regulation of signaling pathways of p38 MAPK and PI3K associated with mature BDNF expression and CREB phosphorylation and subsequently enhanced spatial memory.

【 授权许可】

Unknown   
© Ahn et al. 2015. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.