SUBSCRIBE
Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (1): 68-72.doi: 10.3736/jcim20080113

• Original Experimental Research • Previous Articles     Next Articles

Preliminary assay of p-amyloid binding elements in heart-beneficial recipe

Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu()   

  1. Key State Laboratory of Medical Neurobiology, Shanghai Medical School, Fudan University, Shanghai 200032, China
  • Received:2007-04-05 Online:2008-01-20 Published:2008-01-15
  • Contact: ZHU Cui-qing E-mail:cqzhu@shmu.edu.cn

RichHTML

2

PDF (PC)

71

Like

Objective: To explore whether there are β-amyloid protein (Aβ) binding elements in heart-beneficial recipe (HBR, a compound traditional Chinese herbal medicine), which can ameliorate the cytotoxicity of Aβ.

Methods:The extract of HBR and Aβ1-40 were co-precipitated, and the Aβ1-40 in pellets was detected by immunoblotting. Affi-gel-Aβ1-40 was constructed, and Affi-gel-Aβ1-40 affinity elements from the extract of HBR were analyzed by high-performance liquid chromatography (HPLC). The assay of lactic dehydrogenase (LDH) release from the primary cultured rat cortex neurons was used to evaluate the cytotoxicity of Aβ1-42, and the protection effects of the HBR serum and the Affi-gel-Aβ1-40 treated HBR serum.

Results:Immunoblotting examination showed Aβ1-40 could be co-precipitated with components of HBR following co-incubation, and the amount of Aβ1-40 within pellets decreased when the HBR extract was diluted. Aβ1-40 affinity elements from the extract of HBR, eluted from Affi-gel-Aβ1-40 by glycine solution (pH=2.5), could be detected by HPLC-fluorescent detector system. The analysis of LDH release showed that exposure of neurons to 5 μmol/L Aβ1-42 for 48 h caused a significant increase of LDH release in either a serum free or 10% serum contained culture condition (P<0.01). The rat HBR serum was able to suppress Aβ1-42 induced LDH release (P<0.05), whereas Affi-gel-Aβ1-40 treated HBR serum still maintained the ability to attenuate Aβ1-42 induced LDH release although the effect was somewhat decreased compared with Affi-gel treated HBR serum.

Conclusion:There are Aβ affinity components in HBR, which could not increase the Aβ cytotoxicity, but might be able to inhibit the cytotoxicity of Aβ. The results implied that the exploration of Aβ affinity elements from Chinese medicinal recipe which is effective for Alzheimer disease, might be an important direction in Alzheimer disease therapeutic research area.

Key words: β-amyloid protein, heart-beneficial recipe, lactic dehydrogenase, Alzheimer disease

CLC Number: 

  • R749.16

Figure 1

Co-precipitation of Aβ1-40 with the extracts from heart-beneficial recipe Lane 1 is Aβ1-40 positive marker. The samples of lane 2-4 are pellets precipitated from the solutions containing extract of heart-beneficial recipe (HBR) and Aβ1-40 following incubation. The HBR extract was diluted with PBS by 1∶1 (lane 2), 1∶4 (lane 3) and 1∶8 (lane 4), and co-incubated with Aβ1-40. The Aβ1-40 in centrifuging precipitates was detected by immunoblotting. This experiment was repeated by three times."

Figure 2

HPLC detection of the elements in the heart-beneficial recipe extract absorbed by Affi-gel-Aβ1-40After Affi-gel-Aβ1-40 beads were co-incubated with the extract of HBR, the beads were washed with PBS, and the putative Aβ affinity elements were eluted by glycine solution (pH 2.5). The fluorescent detector in HPLC system could not detect positive signal in the eluents from Affi-gel-Aβ1-40 without OPA derivation (A). After OPA derivation, the positive signals were detected in the eluents from Affi-gel-Aβ1-40 followed by HBR incubation (B, D). However, in the eluents from Affi-gel, as a control sample, only non-specific absorbed elements were detected (C, E)."

Figure 3

Effect of Affi-gel-Aβ1-40 treated heart-beneficial recipe serum on Aβ caused neuronal injuryIn non-serum system (A), the LDH release from primary cultured neurons was significantly increased following 48 h Aβ1-42(1 and 5 μmol/L) treatment. The Sham represented the neurons not treated with Aβ1-42. *P<0.05 and **P<0.01 vs Sham. In a culture system containing 10% serum (B), neurons exposed to Aβ1-42 also caused an increase of LDH release. The serum from HBR administrated rats significantly inhibited Aβ1-42 caused LDH release. And Affi-gel-Aβ1-40 treated HBR serum is still able to inhibit Aβ1-42 cytotoxicity, however it was somewhat less effective than Affi-gel treated HBR serum. *P<0.05 and **P<0.01 vs Aβ1-42 treated normal serum. Each value represents the mean±SD (n=6)."

[1] Selkoe DJ . Alzheimer’s disease: a central role for amyloid. J Neuropathol Exp Neurol. 1994; 53(5):438-447.
doi: 10.1097/00005072-199409000-00003 pmid: 8083687
[2] Wolfe MS . Therapeutic strategies for Alzheimer ' s disease. Nat Rev Drug Discov. 2002; 1(11):859-866.
doi: 10.1038/nrd938 pmid: 12415246
[3] Lin SM. Therapy for Alzheimer disease through regulation of heart and kidney system. Zhongguo Zhong Xi Yi Jie He Za Zhi. 1992; 12(7):393-394. Chinese.
林水淼 . 调理心肾治疗老年性痴呆.中国中西医结合杂志. 1992; 12(7):393-394.
[4] Lin SM , Yang BC. Clinical study on heart-nourishing and brain-tonifying liquid in treating Alzheimer dementia. Shanghai Zhong Yi Yao Da Xue Shanghai Shi Zhong Yi Yao Yan Jiu Yuan Xue Bao. 1996; 10(2):44-47. Chinese with abstract in English.
林水淼, 杨柏灿 . 养心健脑液治疗Alzheimer痴呆的临床研究.上海中医药大学上海市中医药研究院学报. 1996; 10(2):44-47.
[5] Zhou H, Zhao WK. Effect of heart benefitting recipe on the spatial learning and memory impairment and cholinergic system inβ-amyloid protein-treated rats. Zhong Yao Yao Li Yu Lin Chuang. 1998; 14(3):29-31. Chinese with abstract in English.
周晖, 赵伟康 . 调心方对卩A大鼠痴呆模型空间学习记忆障碍和胆碱能系统的影响.中药药理与临床. 1998; 14(3):29-31.
[6] Yang G, Lin SM, Zhao WK , et al. Effect of TX0201 on expression of the apoptosis signal transduction molecule caspase-3 and apoptosis associated genes bcl-2 and bax mRNA in brain tissue of rat analogue model of Alzheimefs disease . Zhongguo Zhong Xi Yi Jie He Za Zhi. 2006; 26(2):147-151. Chinese with abstract in English.
杨戈, 林水淼, 赵伟康 , 等. 调心方有效部位对类AD 大鼠脑组织凋亡信号转导分子Caspase-3及凋亡相关基因bcl-2,bax mRNA表达的影响. 中国中西医结合杂志. 2006; 26(2):147-151.
[7] Yao MZ , Zhao WK. Effect of Tiaoxin Recipe onβ-amy- loid induced neurotoxic model of the primary cultured rat hippocampal neurons. Zhongguo Zhong Xi Yi Jie He Za Zhi. 2001; 21(9):674-676. Chinese with abstract in English.
doi: 10.7661/CJIM.2001.9.674
姚明忠, 赵伟康 . 调心方对β淀粉样蛋白所致原代培养海马神经元细胞毒模型的影响.中国中西医结合杂志. 2001; 21(9):674-676.
doi: 10.7661/CJIM.2001.9.674
[8] Schägger H, von Jagow G . Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kD. Anal Biochem. 1987; 166(2):368-379.
doi: 10.1016/0003-2697(87)90587-2
[9] Li QM, Meng RS, Wei CX, et al. Effects of Naohuan- dan Recipe on learning and memory abilities of SAM-P/ 8 mice and its role in anti-oxidation and anti-apoptosis. Zhong Xi Yi Jie He Xue Bao. 2006; 4(1):48-51. Chinese with abstract in English.
李庆明, 蒙荣森, 魏昌秀 , 等. 脑还丹对快速老化小鼠学习记忆能力的影响及其抗氧化和抗凋亡作用. 中西医结合学报. 2006; 4(1):48-51.
[10] Sadowski MJ, Pankiewicz J, Scholtzova H , et al. Blocking the apolipoprotein E/amyloid-beta interaction as a potential therapeutic approach for Alzheimer's disease. Proc Natl Acad Sci USA. 2006; 103(49):18787-18792.
doi: 10.1073/pnas.0604011103
[11] Goto Y, Niidome T, Akaike A , et al. Amyloid beta- peptide preconditioning reduces glutamate-induced neurotoxicity by promoting endocytosis of NMDA receptor. Biochem Biophys Res Commun. 2006 ; 351(1):259-265.
doi: 10.1016/j.bbrc.2006.10.030
[12] Schwarzman AL, Tsiper M, Greqori L , et al. Selection of peptides binding to the amyloid-protein reveals potential inhibitors of amyloid formation. Amyloid. 2005; 12(4):199-209.
doi: 10.1080/13506120500350762
[1] Umesh Chandra Dash, Atish Kumar Sahoo. In vitro antioxidant assessment and a rapid HPTLC bioautographic method for the detection of anticholinesterase inhibitory activity of Geophila repens. Journal of Integrative Medicine, 2017, 15(3): 231-241.
[2] Lu Yu, Shui-miao Lin, Ru-qian Zhou, Wei-jun Tang, Pin-xian Huang, Ying Dong, Jian Wang, Zhi-hua Yu, Jiu-lin Chen, Li Wei, San-li Xing, Hui-juan Cao, Hong-bin Zhao. Chinese herbal medicine for patients with mild to moderate Alzheimer disease based on syndrome differentiation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(7): 766-776.
[3] Ying-chun Miao , Jin-zhou Tian , Jing Shi , Min Mao. Effects of Chinese medicine for tonifying the kidney and resolving phlegm and blood stasis in treating patients with amnestic mild cognitive impairment: A randomized, double-blind and parallel-controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(4): 390-397.
[4] Cai Lin-lin, Li Hao, Liu Jian-gang, Liu Long-tao, Guan Jie, Liu Ming-fang, Hu Jia, Wei Yun. Effects of early intervention with Huannao Yicong formula effective components on behavior and cholinergic system of β-amyloid precursor protein transgenic mice. Journal of Chinese Integrative Medicine, 2011, 9(3): 292-298.
[5] Yong-chang Diwu, Jin-zhou Tian, Jing Shi. Effects of Chinese herbal medicine Yinsiwei compound on spatial learning and memory ability and the ultrastructure of hippocampal neurons in a rat model of sporadic Alzheimer disease. Journal of Chinese Integrative Medicine, 2011, 9(2): 209-215.
[6] Li-bin Zhan , Xin-ping Niu, Hua Sui, Xiao-yang Gong. Protective effect of spleen-yin-nourishing recipe on amyloid β-peptide-induced damage of primarily cultured rat hippocampal neurons and its mechanism. Journal of Chinese Integrative Medicine, 2009, 7(3): 242-248.
[7] GU Ran, LIU Ru-yu, ZHANG Lan-jiang, HAO Xiao-yan, XIAO Yan, QI Xiao-lan, SHAN Ke-ren, lin REN Xi, LUO Jun, GUAN Zhi-zhong. Protection of Tianshen Yizhi Recipe against low expression of nicotinic receptor and neurotoxicity induced by β-amyloid peptide. Journal of Chinese Integrative Medicine, 2007, 5(5): 564-569.
[8] Rong-fang Ji, Jian-zhao Niu, Shu-qiang Xu, Jian Li, Ji-feng Wang, Jian-ping Liu. Treatment of Alzheimer disease: an evidence-based review. Journal of Chinese Integrative Medicine, 2007, 5(3): 247-254.
[9] Yong Tang, Hai-yan Yin, Fang Zeng, Shu-guang Yu. Pondering in-situ induction of endogenous neural stem cells in hippocampus of rats with Alzheimer disease by acupuncture. Journal of Chinese Integrative Medicine, 2005, 3(5): 351-354.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. An explanation on "guiding principles of clinical research on mild cognitive impairment (protocol)". Journal of Chinese Integrative Medicine, 2008, 6(1): 15-21
[2] Yi-ting He, Qing-lin Zha, Jian-ping Yu, Yong Tan, Cheng Lu, Ai-ping Lv. Principal factor analysis of symptoms of rheumatoid arthritis and their correlations with efficacy of traditional Chinese medicine and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 32-36
[3] Jun Cai, Hua Wang, Sheng Zhou, Bin Wu, Hua-rong Song, Zheng-rong Xuan. Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2008, 6(1): 37-40
[4] Wei Zhang, Xiang-feng Lu, Xiao-mei Zhang, Jian-jun Wu, Liang-duo Jiang. A rat model of pulmonary fibrosis induced by infusing bleomycin quickly through tracheal intubation. Journal of Chinese Integrative Medicine, 2008, 6(1): 60-67
[5] A-gao Zhou, Yong Zhang, Gang Kui, De-Yun Kong, Hai-liang Ge, Qiu-hua Ren, Jia-rong Dong, Sheng Hong, Xu-ming Mao, Yin Wang, Hui-zheng Zhang, Shu-jun Wang. Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 77-82
[6] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[7] Li Zhou, Hong-xing Zhang, Ling-guang Liu, Wen-jun Wan. Effect of electro-acupuncture at Fenglong (GV 16) on nitric oxide and endothelin in rats with hyperlipidemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 89-92
[8] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[9] Xin-jun Wang, Ling-ling Wang . A mechanism of endogenous opioid peptides for rapid onset of acupuncture effect in treatment of depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1014-1017
[10] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
Journal of Integrative Medicine, ISSN: 2095-4964 CN 31-2083/R
Copyright © Journal of Integrative Medicine
Powered by Beijing Magtech Co. Ltd
沪公网安备31010602003416号      沪ICP备18017633号-3