Acetylcholinesterase inhibitory-active components of Rhodiola rosea L.

1. Acetylcholinesterase inhibitory-active components of Rhodiolarosea L. Hua Wang, Gaochao Zhou, Xiangdong Gao, Yidan Wang, Wenbing Yao * School of Life Science and Technology, China Pharmaceutical University, 24 Tongjia xiang, Nanjing 210009, PR China 授課老師：詹于誼 教授 報告者：田仲源、莫博欽

2. Introduction • Rhodiola rosea ("golden root" or "rose root") :景天科(Crassulaceae)紅景天屬(Rhodiola sp.) is a popular plant in traditional medical systems in Eastern Europe and Asia. • 西藏稱“索羅瑪寶 ” • Rhodiola rosea: in a variety of diverse areas of physiological function, including neurotransmitter levels, central nervous system activity, and cardiovascular function.

3. 紅景天的多樣性

4. 生理功能的主要有效成份為 :紅景天苷 (salidroside；C14H20O7) and酪醇(p-tyrosol；C6H10O2) • Application:decreasing depression, enhancing work performance, eliminating fatigue, and preventing high altitude sickness…

5. Acetylcholine

6. Acetylcholinesterase • Acetylcholinesterase (AChE) inhibitors have been widely used in the treatment of Alzheimer’s disease (AD) • The medicinal effect of RR in the treatment of AD is probably mediated by the acetylcholinesterase inhibitory capability of this plant (Wu, Yao, Gao, & Wang, 2003). However, the specific compounds responsible for the anti-cholinesterase activity of RR remain unknown.

7. Alzheimer’s disease (AD) • 1.非正常的老化現象 • 2.喪失記憶和語言障礙 • 3.在美國，阿茲海默症高居成人死因第四位 • 4.神經纖維糾結與斑塊 • 5.膽鹼酯分解酵素抑制劑Cognex 與Aricept • 6.維他命E、阿司匹靈和銀杏

8. BPSD, which includes symptoms such as depression, anxiety, delusions, and agitation, is currently treated with psychotropic medications including antipsychotics, antidepressants, and hypnotics. • Cholinesterase inhibitors may be useful in the management of behavioral and psychological symptoms of dementia (BPSD).

9. Materials and methods • Chemicals Acetylthiocholine iodide (ATCI) as substrate AChE as enzyme 5,5’-dithio-bis-(2-nitrobenzoic acid) (DTNB) • Plant materials the dried rhizome of Rhodiola rosea L.

10. HPLC mobile phases :Solvent A (water) and solvent B (acetonitrile) detector : 280 nm • UV–visible spectrophotometric analysis The sample solution was scanned from 200 to 700 nm • Molecular weight estimation ionization energy was 70 eV; ion source temperature was 230 ℃. The scan range of m/z was 40–800. • NMR spectroscopy

11. Extraction and isolation procedures plant material (200 g) filtered through a filter paper The supernatant was reprecipitated with ethanol centrifugation (3000 rpm, 5 min) boiled ethanol (1 l) for 1 h Filtered extracts evaporated to dryness (46.4 g) centrifuged ( 3000 rpm , 10 min ) remove the remnant gelatine Supernatant (360 ml) was precipited with gelatine filtered through a filter paper Filtered extracts evaporated to dryness (46.4 g) The supernatantwas evaporated to dryness centrifuged ( 3000 rpm ,10 min )to remove tannins boiled ethanol (1 l) for 1 h Extract (40 g) in 400 ml of deionized water

12. supernatant(dryness) H2O chloroform EtOAc n-butanol column chromatography Fractions A36–A59 chloroform/ethyl acetate (1:1) Sephadex LH-20 column 100 fractions (A) methanol/water (1:1) 100 fractions (B) purifier HPLC system compound 1 FractionsB29–B39

13. Bioassay for anti-cholinesterase activity • the colorimetricmethod of Ellman ATCI thiocholine

15. Compare the method of Ellman and Rhee

16. Results