우슬 추출물의 생리활성 연구

Abstract

The dried root of Achyranthes japonica Leveille et Vaniot(Amaranthaceae) is one of the source plants of Achyranthis root. The crude drug is an oriental medicine used for diureties, tonics and as a remedy for blood stasis such arthralgia. Plant of the genus AREE known to contain phytoecdysones, and rich in oleanolic acid glycosides. This study was performed to investigate the effects of ethanol extract of Achyranthis radix(AREE) on antioxidant capacity, lipid metabolism and antithrombogenic capacity The results obtained were as follows : Experiment 1: - Biochemical components of Achyranthis root The biochemical components of Achyranthis root were measured. The crude protein(18.27%), crude fat(2.99%), moisture(16.26%) and carbohydrate (48.48%) were found. Fructose was major in free sugars, lysine in amino acids, linoleic acid in fatty acids, oxalic acid in organic acids, calcium in minerals, potassium ion in actions and chloride ion in anions, respectively. Experiment 2 : - Antioxidant effect of AREE in vitro and in vivo 1. AREE was fractionated by the following solvents : n-hexane, chloroform, ethylacetate and n-butanol. In vitro, antioxidant index of the n-butanol fraction was the highest among fractions of electron donating activity, Rancimat test and activity of nitrite scavenging were the highest at pH 1.2 and the peroxide value for linoleic acid was increased during the storage. 2.The scavenging effects of oxygen free radicals generated in the ethanol- induced hepatotoxicity in the liver of rats were investigated. Sprague-Dawley weighing 100～150g were divided into 6 groups: normal group(NOR), ethanol (35%) treated group(CON), AREE 200 mg/kg treated group(AR-1), AREE 400 mg/kg treated group(AR-2), AREE 200 mg/kg and alcohol treated group(AR-C1), and AREE 400 mg/kg and alcohol treated group(AR-C2). ① The growth rate of the rat was decreased by ethanol administration, however, was gradually increased to a little lower level than the normal group by administering AREE. ② It was also observed that the activities of xanthin oxidase(XO), superoxide dismutase(SOD), catalase and glutathion peroxidase(GSH-Px) in liver that were increased by ethanol were markedly decreased in the AREE administered groups as compared with the CON group. The glutathione(GSH) content in liver was decreased by ethanol adminstration, however, increased after administering AREE. In addition, the value of thiobarbituric acid reactive substances(TBARS) significantly increased in CON group, on the other hand, the administration of AREE reduced TBARS value in liver. Experiment 3 : - Effect of AREE on lipid metabolism in vitro and in vivo 1. Antioxidative activity of AREE against oxidation of LDL was investigated. Oxidation products of LDL were determined by measuring TBARS value. As revealed through TBARS values, n-butanol fraction showed strongest among fractions and decreased LDL oxidation induced by copper ion by 70.6% in comparison with no addition group. 2.AREE(200 mg/kg and 400 mg/kg b.w., p.o.) was administered to rats along with fed high cholesterol diet for 6 weeks and evaluated on lipid metabolism in serum, liver and adipose tissue. We divided into 6 groups: normal group(NOR), high cholesterol diet treated group(CON), AREE 200 mg/kg treated group(AR-1), AREE 400 mg/kg treated group(AR-2), AREE 200 mg/kg and high cholesterol diet treated group(AR-C1), and AREE 400 mg/kg and high cholesterol diet treated group(AR-C2). ① The growth rate of the hyperlipidemic group(CON group) was higher than the normal group, whereas the groups administered AREE were decreased. The feed efficiency ratio(FER) and liver weight in CON group were significantly higher than that of NOR group, but that of CON group with AREE administration was lower than CON group. ② There was a signigicant increase in the activities of serum alanine aminotransferase(ALT), asparate aminotransferase(AST) and alkaline phosphatase(ALP) in the CON group. On the other hand, the administration of AREE decreased ALT, AST and ALP activities in serum. ③ The CON group was increased serum triglyceride, total cholesterol, free cholesterol and LDL-cholesterol levels, and decreased HDL-cholesterol and phospholipid levels as compared with NOR group. AREE administrated groups were increased HDL-cholesterol and phospholipid levels, and decreased serum triglyceride, total cholesterol, free cholesterol, ratio of cholesteryl ester and LDL-cholesterol levels as compared with CON group. AREE increased HDL-C/T-C ratio and lowered atherogenic index. The hepatic and adipose tissue contents of total lipid, total cholesterol and triglyceride were also lowered in AREE administrated groups than CON group. ④ The activities of HR-LPL and TE-LPL in mesenteric and epididymal adipose tissues were increased in CON group as compared with NOR and administrated AREE groups. CON group also showed higher LPL mRNA abundance in both mesenteric and epididymal adipose tissues than NOR and administrated AREE groups. ⑤ The LPL activity in adipose tissue was positive correlated with the triglyceride concentration in serum, but negative correlated with HDL-cholesterol concentration in serum. Experiment 4 : - Antithrombogenic effects of AREE in vitro and in vivo 1. AREE was found to have fibrinolytic activity in vitro. 2. The antithrombogenic effects of AREE on pulmonary embolism by collagen and epinephrine in mice and coagulation parameters in endotoxin-induced disseminated intravascular coagulation rats in vivo were also investigated. ① AREE administered groups prevented death due to the formation of pulmonary enbolism by collagen and epinephrine in mice. ② The prothrombin time, activated partial thromboplastine time and blood clotting time in rats were significantly shortened in AREE administered groups as compared with CON group. AREE administered groups increased fibrinogen level and hematocrit value as compared with CON group in thrombus model induced by endotoxin. In conclusion, these results suggest that AREE has a possible positive effect on the liver function in hepatotoxicity-induced rats. In addition, AREE is believed to be a possible protective or curative effects for fatty liver and hyperlipidemia-induced by a high cholesterol diet and is also suppressive activity for a blood coagulation system.