Study On Optimization Of Purification Process And Pharmacodynamics Of Total Organic Acids From The Roots Of Aralia Cordata Thunb.(?)

Aralia cordata Thunb.is the roots and rhzomes of Araliaceae AraliaIt is spicy and bitter in taste,while it is tepid in nature.This plant is mainly used for the treatment of phlegm resistance collaterals,lumbar muscle strain,liver and kidney deficiency and other syndromes because of its efficacy on dispelling wind,relieving swelling and pain,dehumidifying and activating blood.Aralia cordata Thunb.was recorded firstly as a medicinal product in Compendium of Materia Medica.In this paper,studies on optimization of purification process of total organic acids,contents determinations of kaurenic acid and pimaric acid,pharmacodynamics on anti-inflammatory and analgesic,antibacterial,antitumor and mechanism of anti-inflammatory with rats induced by LPS were investigated.The main contents and results are as follows:In the first part,investigating optimization of purification process of total organic acids based on preliminary research.The content of total organic acids was established by the acid-base titration method,then made the further optimized by TLC-silica gel column method and recrystallization and decolorization method.Finally,we choose petroleum ether(60?)-ethyl acetate(80:1)as the best elution condition,the contents of total organic acids about Aralia cordata Thunb.and optimization are respectively 3.4%,96%.In the second part,studying the contents determinations of kaurenic acid and pimaric acid from the roots of Aralia cordata Thunb.by HPLC-DAD.Then we find that the determinations of kaurenic acid and pimaric acid are respectively0.4739%,0.5923%.In the third part,studying the contents determinations of kaurenic acid and pimaric acid about the optimized total organic acids by HPLC-DAD.Then we find that the determinations of kaurenic acid and pimaric acid are respectively35.94%,48.98%.In the fourth part,investigating on anti-inflammatory and analgesic about theoptimized total organic acids,on antibacterial,anti-tumor and mechanism of anti-inflammatory with rats induced by LPS about kaurenic acid,pimaric acid,kaurenic acid:pimaric acid(4:5)and optimized total organic acids.Studying the anti-inflammatory and analgesic effects by five classical models those are xylene-induced mouse ear swelling method,egg white-induced rat paw edema method,hot plate stimulation method,acetic acid-induced mouse writhing method,formaldehyde-induced mouse paining method;evaluating the antibacterial activity with Escherichia coli,Staphylococcus aureus,Enterobacter cloacae,Klebsiella pneumoniae Staphylococcus epidermidis and Pseudomonas aeruginosa by dichotomanthes tube;studying the anti-tumor activity wirh human leukemia cell line MV4-11,human breast cancer cell line MCF-7 and MDA-MB-231 by MTT method;investigating the effective mechanism of anti-inflammatory with rats induced by LPS,testing the expressions of IL-1?,TNF-? and NO via by Elisa and Griess method while testing the levels of JNK and p-JNK in lung tissue by Western blot method.The results show that the optimized total organic acids has a favorable effect on anti-inflammatory and analgesic;kaurenic acid,pimaric acid,kaurenic acid:pimaric acid(4:5)and optimized total organic acids all make certain antibacterial activities,especially on Pseudomonas aeruginosa;those compitions all have a significant inhibitory effect on human leukemia cell line MV4-11 but no effects on human breast cancer cell line MCF-7 and MDA-MB-231;expressions of IL-1?,TNF-?and NO are obviously reduced in serum treated with kaurenic acid:pimaric acid(4:5),meanwhile,pimaric acid makes a significant inhibitory effect on TNF-?,but all of their cytokines are not obviously reduced in lung tissue treated with above all compositions,in addition,the level p-JNK can be inhibited in different degrees by all compositions in which the optimized total organic acids is the best,so it indicates that the total organic acids from Aralia cordata Thunb.have a good anti-inflammatory activity.