Anti-atheroscleortic Effects of Herbal Extracts

探討藻藍素和蕃茄紅素降血脂及抗氧化之分子機轉吳介信張永賢中國醫藥大學 (臺灣)中图分类号：R931.71 文献标识码：A 文章编号：1818－0086（2008）02

Anti-atheroscleortic Effects of Herbal Extracts A. 正常兔子之主動脈 B. 餵食0.5%高膽固醇的兔子主動脈 C. 2%山藥及0.5%膽固醇 D. 300mg/kg厚朴葛根及澤瀉 E. Probucol

藍綠藻 • 1921年生物學家在非洲的史瓦濟蘭發現目前已知地球上最早出現的生物:藍綠藻(Cyanobacteria, blue-green algae） • Spirulina sp. • 螺旋藻屬於藍藻門,紐子目,螺旋藻科,搖藻屬 • 生長於鹼性鹽湖

螺旋藻主要組成 • C-藻藍素(C-phycocyanin,CPC) • Vitamin B12 • 礦物元素: Ca, Zn, Fe, K • β-carotene • Chlorophyll a • γ-Linolenic acid (GLA)

Table 1.螺旋藻於各種動物及人類疾病模式上的研究

Male Golden Syrian hamsters 7 weeks old Normal diet (N) 0.2% cholesterol+ 5% soybean oil 0.2% cholesterol High cholesterol diet (H) 1% Lopid + High cholesterol diet (L) Low dose spirulina powder or tomato powder + High cholesterol diet High dose spirulina powder or tomato powder +High cholesterol diet 8 weeks Sacrifice Blood Liver

Blood ＊lipid determination --Serum levels of LDL ,HDL,TG,TC, ＊Determination of anti-oxidative effect --serum MDA assay --LDL isolation and conjugated diene determination Liver ＊proteomics --2-dimensional polyacrylamide gel electrophoresis, 2D-PAGE --LC-MS/MS analysis

八週高膽固醇餵食、給予spirulina powder對hamster生長情況影響 Each value represents mean±SEM (n=6) ＊significantly different (p<0.05) when compared with cholesterol group by student’s t test. ＊＊significantly different (p<0.01) when compared with cholesterol group by student’s t test.

Table 1: Serum total cholesterol (TC), triacylglycerol (TG), low density lipoprotein (LDL), high density lipoprotein (HDL), GOT, GPT levels after 8 weeks treatment periods. Each value represents mean±SEM (n=6) ＊significantly different (p<0.05) when compared with cholesterol group by student’s t test. ＊＊significantly different (p<0.01) when compared with cholesterol group by student’s t test. ＊＊＊significantly different (p<0.001) when compared with cholesterol group by student’s t test. 0.5% spirulina powder contained 15mg/kgbw C-PC. 5% spirulina powder contained 150mg/kgbw C-PC.

Table 2: Serum total cholesterol (TC), triacylglycerol (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) levels after 8 weeks treatment periods. Each value represents mean±SEM (n=6) ＊significantly different (p<0.05) when compared with cholesterol group by student’s t test. ＊＊significantly different (p<0.01) when compared with cholesterol group by student’s t test. ＊＊＊significantly different (p<0.001) when compared with cholesterol group by student’s t test. 3% tomato powder contained 3.3mg/kgbw lycopene. 9% tomato powder contained 9.9mg/kgbw lycopene.

Serum MDA assay MDA (malondialdehyde) + TBA(2-thiobarbituric acid) H+ ∆ Thiobarbituric acid-reactive substances (TBARS) (red color) O.D 540 nm lipid peroxidation cascades (Philip C.Burcham1,1998)

＊＊ Fig.1 Amounts of serum MDA of hamsters fed with various diets in animal experiments. Each value represents mean±SEM (n=6) ＊significantly different (p<0.05) when compared with cholesterol group by student’s t test.

＊＊＊ ＊＊＊＊＊＊ Fig.1 Amounts of serum MDA of hamsters fed with various diets in animal experiments. Each value represents mean±SEM (n=6) ＊＊＊significantly different (p<0.001) when compared with cholesterol group by student’s t test.

LDL isolation and conjugated diene determination a is the lag phase in minutes The tangent b gives the maximal rate of oxidation d-c gives the maximum concent- ration of conjugated diene before onset of decompositions c-e gives the rate of decomposition 1.Lag phase 2.Propagation phase 3.Decomposition phase 3 2 1 Monitoring of Cu2+-mediated LDL oxidation by the conjugated diene method. (Puhl and others 1994)

Table3. Lag time of serum LDL oxidation of hamsters fed with various diets in animal experiment. Each value represents mean±SEM (n=3) ＊significantly different (p<0.05) when compared with cholesterol group by student’s t test.

Groups Cholesterol 3% tomato powder 9% tomato powder lag time (min) 208.33±22.05 321.67±43.43 398.33±9.28*** Table4. Lag time of serum LDL oxidation of hamsters fed with various diets in animal experiment. Each value represents mean±SEM (n=3) ＊＊＊significantly different (p<0.001) when compared with cholesterol group by student’s t test.

2-dimensional polyacrylamide gel electrophoresis,2D-PAGE analysis Steps in 2-D PAGE

From http://binfo.ym.edu.tw/binfo/01s/pro_info.htm

LC-MS/MS analysis of cholesterol-induced protein expression

LC-MS/MS analysis of cholesterol-induced protein expression Adenylate Kinase isozyme 2 Carbonic Anhydrase III

LC-MS/MS analysis of cholesterol-induced protein expression N C 9% TP CAIII AK2 b-actin

Anti-atheroscleortic Effects of Herbal Extracts