1 193 J Chin Med 14(3): 193-203, 2003 1 2 2 2 4 5 3 1 3 5 4 2 (2003 6 10 2003 7 28 2003 8 14 ) 2,2-diphenyl-1-picrylhydrazyl DPPH 78 µg/ml DPPH 0.1 µg/ml Vitamin C DPPH 8 mg/ml Vitamin E ICR 04-22052121 4163 04-22032798 E-mail: hsumd736@ms67.hinet.net
194 2 2002 26 % 41 % 29 % 113 % 3 17 β-estrodiol 4 5 6 7 8 9 10 11 A C E 12 13 Auricularia auricula Glycine max Fructus Ziziphi Jujubae ICR 21-24 12 25 ± 1 C 60 ± 5 %
195 DPPH (2,2-diphenyl-1-picrylhydrazyl), Sigma Chemical Co., USA; 1,2-diacyl-sn-glycero-3-phosphocholine, Sigma Chemical Co., USA; 2-thiobarbituric acid (TBA), Sigma Chemical Co., USA, 0.7 % in NaOH solution; butylated hydroxytoluene (BHT), Sigma Chemical Co., USA, 2 % in ethanol; ferric chloride (FeCl 3 ), Riedl-deHaen; trichloroacetic acid (TCA), Riedl-deHaen; 17-α-ethinyl estradiol (EE), Sigma Chemical Co., USA 1,2-diacyl-sn-glycero-3-phosphocholine 10 mg/ml liposome solution 20 mm sodium phosphate buffer ph 7.4 30 3 45 mg 17-α-ethinyl estradiol 1 ml 95 % 0.45 µg/ml 2.25 g 0.75 g 3 g 40 ml 100 C 30 150 ml 1 Blois 14 DPPH 100 µl 2 Vitamin C 100 µl 0.2 mm DPPH 30 517 nm DPPH scavenging effect % = 1- / 100 250 ìl ddh 2 O 1 ìl 0.5 M FeCl 3 50 ìl 125 mm L-ascorbic acid 700 µl 37 C 2 400 ìl 30 % TCA 50 µl 2 % BHT 250 µl 0.7 % TBA 400 ìl 95 C 50 1 mln-butanol 532 nm TBARS thiobarbituric acid reactive substances ( ) 150 ml 1 3
196 0.0026 1 8 mg 10 80 mg ICR 21-24 9 3 A B 10 µg/kg 0.45 µg/ml 0.2 ml C 1 20 mg/ml 0.2 ml D 10 200 mg/ml 2 0.2 ml 10 ml/kg 15 4 / Kruskal-Wallis H test Dunnett t Test p < 0.1 ( )
197 DPPH scavenging effect % 100 L-Ascorbic acid C.Q. tea 75 50 25 0 0.001 0.01 0.1 1 10 100 1000 Concentration(mg/ml) 1 DPPH vitamin C N = 3 mean SD DPPH scavenging effect % 75 50 25 0 CK tea Black bean Fructus Jujubae Wood ear 0.0625 0.25 1 4 Concentration (x 10-2 crude extract) 2 DPPH N = 3 mean SD TBARS Absorbance (532 nm) 1.00 0.75 0.50 0.25 0.00 control Vitamin E C.Q. tea 0 2 4 6 8 10 Concentration (mg/ml) 3 a-tocopherol Vitamin E vitamin E N = 3 mean SD
198 (A) Uterus wet weight (mg) 20 15 10 5 * 0 C EE 1x C.Q. 10x C.Q. (B) * * (uterus wet weight/ body weight) *100 0.15 0.10 0.05 0.00 C EE 1x C.Q. 10x C.Q. 4 ICR * p < 0.1 C EE 1 C.Q. 10 N = 5 mean SD 4B p < 0.1 H & E 200 1 3 5 400 400 0 1 400 1 6
199 5 H & E stain 200 (A) (B) (C)1 (D)10 Ep 6 H & E stain 400 (A) (B) (C) Mit
200 16 A C E 17 18-20 13 DPPH DPPH 517 nm DPPH DPPH DPPH DPPH 78 µg/ml 88.97 % Vitamin C 171 ng/ml 82.01 % DPPH DPPH 21 16 Raloxifene 22 ICR
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203 J Chin Med 14(3): 193-203, 2003 ANTIOXIDATIVE EFFECTS AND ESTROGENIC ACTIVITY OF CHINESE HERBS CHARMING-QUEEN TEA Shan-Yu Su 1, Chih-Hsiung Lee 2, Cheng-Wei Lai 2, Ying-Yuand Wang 2, Min-Chien Lai 4, Ming-Tsung Yu 5 and Ching-Hsaing Hsu 3 1 Department of Chinese Medicine 5 Institute for integration of Traditional Chinese and Western Medicine 3 Department of Pediatric, China Medical University Hospital, Taichung, Taiwan 4 Department of Pathology, China Medical University Taichung, Taiwan 2 GenMont Biotechnology Incorportation Tainan, Taiwan (Received 10 th June 2003, revised Ms received 28 th July 2003, accepted 14 th August 2003) Supplement of estrogens is conventional treatment for climacteric women to release postmenopausal symptoms. However, plenty of studies reviewed that traditional hormone replacement therapy induced many side effects. Our specific aim in this study is to look for Chinese herbs with estrogen-like activity. This study detected antioxidative effects and estrogenic activity of extract from Charming-Queen tea (C.Q. tea) which composed of fructus jujubea, black bean and Jew s ear. The antioxidative study showed dose dependence of C.Q. tea concentration with scavenging effect of 2,2- diphenyl-1-picrylhydrazyl (DPPH) free radicals and the protective effect of liposomes from ferric chloride. At the concentration of 78 µg/ml, C.Q. tea acted similar scavenging effect as 0.1 µg/ml Vitamin C. The DPPH scavenging activity of C.Q. tea is higher than any of the single fructus jujubea, black bean or Jew s ear. At the concentration of 8 mg/ml, C.Q. tea acts the similar liposome protective activity as Vitamin E. Estrogen-like activity was determined via uterotrophic assay. After feeding immature ICR mice with C.Q. tea, both uterine weights and body weights were measured. The results showed that ten times C.Q. tea can show an increase in uterine wet ratio. In addition, uterine biopsy showed higher concentration of C.Q. tea can increase the thickness of epithelium and the numbers of mitotic cells but lower concentration of C.Q. tea didn t. We therefore concluded that C.Q. tea have antioxidative effect and estrogen-like effect in the animal model. Key words: C.Q. tea, Antioxidation, Phytoestrogen. Correspondence to: Ching-Hsaing Hsu, Allergy, Immunology and Rheumatology section, Department of Pediatric, China Medical University Hospital, Tel: 04-22052121 ext. 4163, Fax: 04-22032798, E-mail: hsumd736@ms67.hinet.net