Inhibitory Effect Of Radix Angelicae Pubescentis On Active Ingredients Of Cholinesterase

TLC-bioautographic assay and Micro plate high-throughput screening assay are used to investigate the effect of51natural product samples on the activities of Acetylcholinesterase (AChE) and Butyrylcholinesterase (BuChE) and are screened for the best inhibitors of both AChE and BuChE in this paper,Under the concentration of lmg/ml, this study screened11inhibitors for both AChE and BuChE from51natural product samples. The AChE and BuChE percentage inhibition of three traditional Chinese medicines (Radix Angelicae Pubescentis, Cnidium monnieri (L.)Cuss and Peucedanum praeruptorum Dunn)were32.22±4.32%,31.8±0.72%;23.33±3.51%,32.09±4.62%;22.00±2.00%,47.70±1.13%,respectively.The total ChE percentage inhibition of these three traditional Chinese medicines were more than50%. Radix Angelicae Pubescentis, Cnidium monnieri (L.)Cuss and Peucedanum praeruptorum Dunn showed the maximum inhibitory potential.We consulted some relevant references of Radix Angelicae Pubescentis, Cnidium monnieri (L.)Cuss and Peucedanum praeruptorum Dunn,we have found that the alcohol extracts of Radix of Angelicae Pubescentis showed significant cognitive benefits in AD rats,but the activity compounds did not reveal at all.So Radix Angelicae Pubescentis was selected in this paper.Angelica pubescens Maxim.f.biserrata Shan et Yuan belongs to the Apiaceae family grows in Chinese provinces such as Hubei and Sichuan, The radix of Angelicae Pubescentis is used in Traditional Chinese Medicine as herbal drug Duhuo.Coumarins and furanocoumarins are the main constituents of this drug.In Traditional Chinese Medicine, Duhuo is used to treat pain, fever, rheumatism, heaviness of the low-back and knees, and contracture of the limbs. Some recent investigations have shown the anti-inflammatory, antioxidant, antitumor activity of this plant, furthermore, recent study reveals that the alcohol extracts of Radix of Angelicae Pubescentis showed significant cognitive benefits in AD rats.This paper reviews the main chemical components and pharmacological activities of Radix of Angelicae Pubescentis in recent30years.TLC-bioautographic assay and Micro plate high-throughput screening assay are used to investigate the effect of the different extracts of Radix Angelicae Pubescentis on the activities of Acetylcholinesterase (AChE) and Butyrylcholinesterase (BuChE) in this paper. The part of volatile oil had positive result on the TLC-bioautographic assay and Microplate high-throughput screening assay. Under the concentration of1μl/ml, the AChE and BuChE percentage inhibition were92.38±0.81%,63.74±1.48%. The IC50of volatile oil on inhibition of AChE and BuChE were0.21±0.006μl/ml,0.47±0.30μl/ml, respectively. In the part of non-volatile, the ethanol extract, petroleum ether extract and chloroform extract from Radix Angelicae Pubescentis had positive result on the TLC-bioautographic assay and Microplate high-throughput screening assay. Under the concentration of1mg/ml, the AChE percentage inhibition of these3parts were16.88±0.42%,11.82±0.55%,44.33±3.21%, respectively; the BuChE percentage inhibition were13.97±1.17%,14.64±1.10%,18.63±1.24%, respectively. The ethanol extract, petroleum ether extract and chloroform extract from Radix Angelicae Pubescentis had inhibitory effect on both AChE and BuChE.The chloroform extract showed the maximum inhibitory potential. So this article describes the dual-cholinesterase inhibitory activity of volatile oil and non-volatile oil of Radix Angelicae Pubescentis and the bioguided isolation and identification of dual-ChE inhibitory compounds from the radix chloroform extracts.Some modem separation technologies and methods are used to separate the chloroform extract of Radix Angelicae Pubescentis, such as conventional methods, Sephadex LH-20column chromatography, HPLC, etc.12compounds were separated from the radix chloroform extracts.By the methods of classical chemistry and a variety of advanced spectroscopic techniques (including UV,1HNMR,13CNMR), we identified the structure of8compounds which are Osthole(RAPI), Columbianetin(RAPII), Columbianetinacetate(RAPIII),8-Methoxypsoralen(RAPIV), Columbianadin(RAPV), bisabolangelone(RAPVI), umbelliferone(RAPVIII), angelol M(RAPIX). The other4compounds including RAPVII, RAPX, RAPXI, RAPXII are being identified. In the part of the non-volatile,10compounds from the radix chloroform extracts are screened by Micro plate high-throughput screening assay, all of which have the effect on inhibition of ChE. Osthole and Columbianetin were proven to achieve AChE and BuChE inhibition at a rate of over50%at a concentration of1mg/ml, the AChE and BuChE percentage inhibition of Osthole were90.25±2.67%,68.50±2.71%. The IC50of Osthole on inhibition of AChE and BuChE were489.94±22.12μM,1312.34±53.62μM,respectively. The AChE and BuChE percentage inhibition of Columbianetin were59.67±2.27%,39.32±1.67%. The IC50of Columbianetin on inhibition of AChE was1238.91±79.49μM. The ChE inhibitory activity of Galanthamine which is a positive drug for AD was reported and the IC50values of AChE and BuChE were0.37μM and8.38.3μM, respectively.The result of this study indicated that the parts of non-volatile oil showed the weak inhibitory effect of ChE activity. The experiments in vitro were only finished in this study, so it is necessary to carry out experiments in vivo and experiments of Pharmacokinetics, the absorption of the blood in the body are needed to have some further more research.