飼養高脂飼料 15 週後犧牲，將胸主動脈組織震碎，抽取組織蛋白質，進行西 方轉漬法測試發炎因子 MMP-9 及 IL-1β蛋白質表現量。圖 3-21A、3-22A 為顯影後 代表圖，MMP-9 及 IL-1β分子量分別為92kDa、17kDa，其表現量由 Image J 量化後，
數值以 MMP-9、IL-1β表現量/β-actin 表現量表示(圖 3-21B、3-22B)。結果發現，胸 主動脈中 MMP-9 及 IL-1β蛋白質表現量四組間皆無顯著差異(圖 3-21B、3-22B)
表 3-2 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週體重增加量、平均 攝食量及攝食效應
Table 3-2. The body weight gain, food intake and feed efficiency of ApoE knockout mice fed different diets for 15weeks
組別 假手術 (n=9)
控制組 (n=12)
苜蓿 EA1%
(n=12)
茉莉甲醇 萃物 1%(n=11)
Sham OVX OVX+A OVX+J
Initial body weight (g) 16.1±1.5 17.0±1.5 17.0±1.9 16.0±1.5 Final body weight (g) 23.2±2.3 ab 25.8±3.7 b 22.9±3.2 a 23.4±3.7 ab
Weight gain(g) 7.09±1.56 ab 8.79±3.90 b 5.90±2.94 a 7.46±3.55 ab
Food intake (g/mice/day) 2.47 2.43 2.46 2.25 Feed efficiency
(g/100g diet) 2.93 3.45 2.28 3.16
1. Feed efficiency = weight gain (g)/total food intake (100g) 2. Values are means ± S.E.M.
3. Data were analyzed by one-way ANOVA and LSD multiple comparison test. Values not sharing the same letter are significantly different at p<0.05.
表 3-3 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週後組織絕對及相對重量 Table 3-3. Absolute and relative organ weights in ApoE knockout mice fed test diets for 15 weeks 腎周脂 (g) 0.117±0.044 0.235±0.177 0.161±0.150 0.139±0.104
% of BW 0.232±0.299 0.656±0.682 0.593±0.585 0.426±0.434 子宮周脂+腎周脂 (g) 0.286±0.105 a 0.722±0.486 b 0.496±0.471 ab 0.390±0.348 ab
% of BW 1.29±0.42 a 2.78±1.53 b 2.08±1.53 ab 1.65±1.25 a 肝 (g) 1.020±0.143 ab 1.132±0.202 a 0.982±0.201 ab 0.937±0.174 b
% of BW 2.74±2.059 3.483±1.838 3.29±1.699 3.157±1.728 腎 (g) 1.137±0.163 b 1.144±0.624 b 0.702±0.398 a 1.077±0.249 b
% of BW 2.972±2.331 3.178±2.607 2.092±1.823 3.583±2.053 脾 (g) 0.152±0.088 0.151±0.058 0.173±0.090 0.184±0.056
% of BW 0.301±0.360 0.375±0.312 0.414±0.311 0.587±0.568 心 (g) 0.137±0.019 ab 0.142±0.022 b 0.125±0.017 a 0.138±0.019 ab
% of BW 0.287±0.337 0.416±0.312 0.381±0.298 0.460±0.376
1. Feed efficiency = weight gain (g)/total food intake (100g) 2. Values are means ± S.E.M.
3. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
Values not sharing the same letter are significantly different at p<0.05.
表 3-4 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週實驗期間禁食血液 生化值及犧牲後肝脂濃度
Table 3-4. Fasting serum and Hepatic Cholesterol (TC) and triacylglycerol (TG) concentrations in sham-operated or ovariectomized (OVX) apoE knockout mice fed the experimental diets for 15 weeks
Total Cholesterol (mg/dl)
0 week 227.9±74.6 229.0±111.3 216.5±43.3 247.5±86.5 4 week 505.1±100.5 498.0±203.6 479.9±145.2 467.8±83.7 8 week 511.3±102.2 539.1±196.3 429.9±98.2 443.8±123.8 15 week 516.2±161.9 b 444.4±181.2 ab 353.3±67.5 a 451.9±67.6 ab Triglyceride (mg/dl)
0 week 81.3±25.3 76.6±16.0 71.8±28.2 74.9±16.0 4 week 59.1±12.5 63.7±12.9 62.0±15.4 62.7±16.0 8 week 62.5±21.5 71.7±28.1 67.1±22.5 76.6±29.3 15 week 68.0±13.2 65.4±21.3 76.5±11.0 78.1±15.8 Glucose (mg/dl)
0 week 75.4±13.0 76.8±14.2 85.9±18.4 72.7±11.8 4 week 80.7±12.8 88.1±25.7 89.5±18.3 82.6±10.3 8 week 81.2±20.5 95.1±17.8 88.3±14.7 76.6±29.3 15 week 98.8±27.5 89.5±26.9 84.4±30.5 78.1±15.8
Insulin (μg/l) 1.16±0.46 1.23±0.58 1.29±0.69 1.55±0.67
NEFA (mmole/l) 1.95±0.44 1.72±0.70 1.80±0.90 1.78±0.75
HOMA-IR 7.00±4.15 6.67±4.00 6.44±3.34 8.35±6.33
Adiponectin(μg/ml) 4.62±1.48 4.96±2.48 4.85±2.95 5.54±2.34
Leptin(ng/ml) 3.79±0.38 a 7.83±5.58 b 7.36±4.46 ab 6.95±4.22 ab 1. Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD. Values not
sharing the same letter are significantly different at p<0.05.
2. HOMA-IR index = insulin（μU/mL）× glucose（mmol/L）/22.5
表 3-5 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週犧牲後肝脂濃度 Table 3-5. Hepatic Cholesterol (TC) and triacylglycerol (TG) concentrations in sham-operated or ovariectomized (OVX) apoE knockout mice fed the experimental diets for 15 weeks
假手術 1.Values are Mean±SEM
2. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
Values not sharing the same letter are significantly different at p<0.05.
表 3-6 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週後腎臟脂肪濃度 Table 3-6. The renal cholesterol (TC) and triacylglycerol (TG) concentrations in
ApoE knockout mice in sham-operated or OVX fed a high-fat diet fed test diets for 15 weeks
假手術 kidneyTG (mg/g) 8.74±1.20 8.96±2.34 8.93±4.85 6.93±1.36 1.Values are Mean±SEM
2. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
Values not sharing the same letter are significantly different at p<0.05.
表 3-7 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週後頭臂動脈橫切面 之新生內層及中層面積比率 Intima/Media Ratio
Table 3-7. The I/M ratio of innomenate in ApoE knockout mice in sham-operated or OVX fed a high-fat diet fed test diets for 15 weeks
Intima/Media Area ratio 1.44±1.22 1.51±0.96 0.71±0.78 0.73±0.96
(p=0.063)
1.Values are Mean±SEM
2. Data were analyzed by Mann-Whitney test.
表 3-8 ApoE 剔除去卵巢及假手術小鼠餵食實驗飼料 15 週後脂肪細胞大小 Table 3-8 . The size of adipocyte in ApoE knockout mice in sham-operated or OVX fed a high-fat diet fed test diets for 15 weeks
1.Values are Mean±SEM
2. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
Values not sharing the same letter are significantly different at p<0.05.
Sham, n=9; OVX+A, n=12; OVX+JM, n=11; OVX, n=12
圖 3-1 假手術或卵巢切除之 ApoE 剔除小鼠餵食實驗飼料 15 週生長曲線 Fig 3-1 The growth curve of ApoE knockout female mice fed high fat diets
(Sham & OVX), high fat diets supplemented with 1% alfalfa EAE (OVX+A) or 1% Jasaminum (OVX+J) for 15 weeks.
All values are means ± S.E.M. Data were analyzed by one-way-ANOVA and LSD multiple comparison test
圖 3-2 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後血液 insulin 濃度 fig.3-2 Serum level of insulin in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-3 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後血液 NEFA 濃度 fig.3-3 Serum level of NEFA in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-4 假手術及卵巢切除 ApoE 剔除鼠餵食實驗飼料 15 週後肝臟中總膽固醇及 三酸甘油酯濃度
Table 3-4 Hepatic total cholesterol(TC) (A) and triacylglycerol(TG) (B)concentrations
in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks1
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test. Values not sharing a common superscript letter among Sham, OVX , OVX+A, OVX+J, groups are significantly different at p< .05 level.
A
B
圖 3-5 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料第 0 至 15 週血液膽固醇濃度 fig.3-5 Serum level of total cholesterol in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. mg/dl. Data were analyzed by one-way ANOVA and LSD multiple comparison test. Values not sharing a common superscript letter among Sham, OVX, OVX+A, OVX+J groups are significantly different at p< .05 level.
圖 3-6 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料第 0 至 15 週三酸甘油酯濃度 fig.3-6 Serum level of triglyceride in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-7 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料第 0 至 15 週血液葡萄糖濃度 fig.3-7 Serum level of glucose in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-8 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後血液 adiponectin 濃度 fig.3-8 Serum level of adiponectin in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-9 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後血液 leptin 濃度 fig.3-9 Serum level of leptin in sham-operated or ovariectomized (OVX)
ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. ng/ml. Data were analyzed by one-way ANOVA and LSD multiple comparison test.
圖 3-10 假手術及卵巢切除 ApoE 剔除鼠餵食實驗飼料 15 週後腎臟中總膽固醇及 三酸甘油酯濃度
Table 3-10 Renal total cholesterol (TC) (A) and triacylglycerol (TG) (B) concentrations in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks
Values are means ± S.E.M. mg/dl. Data were analyzed by one-way ANOVA and LSD multiple comparison test. Values not sharing a common superscript letter among Sham, OVX, OVX+A, OVX+J groups are significantly different at p< .05 level.
A
B
圖 3-11 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後頭臂動脈 血管 Intima / Media area ratio
Fig.3-11 intima/media ratio of brachiocephalic artery in sham-operated or
ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
Values are means ± S.E.M. Data were analyzed by Mann-Whitney test.
圖 3-12 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後蘇木精-伊紅染色之 肝臟切片(scale bar=100μm)
Fig 3-12 The lipid accumulation in the liver sections stained by hematoxylin-eosin of sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.(scale bar=100μm)
圖 3-13 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後 periodic acid-Shiff (PAS)染色染色之腎臟切片
Fig 3-13 The kidney sections stained by periodic acid-Shiff (PAS) of sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
圖 3-14 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後頭臂動脈 血管切片(蘇木精-伊紅染色) (scale bar=100μm)
Fig 3-14 The brachiocephalic artery stained by hematoxylin-eosin of sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15weeks.(scale bar=100μm)
主動脈脂肪堆積情形(1 小格為 1mm) The deposition of lipid in the aorta
圖 3-15 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後主動脈損傷情形 Fig.3-15 Atherosclerotic lesion formation in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
The aortic arch were dissected out and examined for atherosclerotic lesion formation using the en face technique. A, four representative aorta from the Sham, OVX, OVX+A and OVX+J group. B, the extent of atherosclerotic lesion formation in each mouse, expressed as percentage of aortic surface area covered by atherosclerotic lesions. The mean atherosclerotic lesion area of each group of mice is indicated by a horizontal bar with mean value shown next to it.
圖 3-16 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後脂肪切片(蘇木精 -伊紅染色)
Fig 3-16 The adipocyte stained by hematoxylin-eosin of sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks.
圖 3-17 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後對於頭臂動脈動脈 表現 MMP-9 之影響
Fig 3-17 Immunohistochemical analysis of MMP-9 in brachiocephalic artery in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks. (scale bar=100μm)
圖 3-18 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後對於頭臂動脈動脈 表現 IL-1β之影響
Fig 3-18 Immunohistochemical analysis of IL-1β in brachiocephalic artery in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks. (scale bar=100μm)
圖 3-19 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後對於頭臂動脈動脈 表現 TM 之影響
Fig 3-19 Immunohistochemical analysis of TM in brachiocephalic artery in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks. (scale bar=100μm)
圖 3-20 假手術或卵巢切除之 ApoE 剔除鼠餵食實驗飼料 15 週後對於頭臂動脈動脈 表現 HO-1 之影響
Fig 3-20 Immunohistochemical analysis of HO-1 in brachiocephalic artery in sham-operated or ovariectomized (OVX) ApoE knockout mice fed the experimental diets for 15 weeks. (scale bar=100μm)
圖 3-21 苜蓿芽乙酸乙酯萃物及茉莉甲醇萃物對 ApoE 剔除鼠胸主動脈中 MMP-9 蛋白質表現量的影響
Fig 3-21 Effects of ethyl acetate extracts of alfalfa and methanol extracts of jasminum on the protein expression of MMP-9 in thoracic aorta.
Tissue protein was collected for MMP-9 analysis using western blots. Amounts of
MMP-9 were quantified using β-actin as internal control. Values are means ± S.D, Sham group n=8, OVX group n=8, OVX+A group n=6, OVX+J group n=8, Data were analyzed by one-way ANOVA test.
圖 3-22 苜蓿芽乙酸乙酯萃物及茉莉甲醇萃物對 ApoE 剔除鼠胸主動脈中 IL-1β 蛋白質表現量的影響
Fig 3-22 Effects of ethyl acetate extracts of alfalfa and methanol extracts of jasminum on the protein expression of IL-1β in thoracic aorta.
Tissue protein was collected for IL-1β analysis using western blots. Amounts of IL-1β were quantified using β-actin as internal control. Values are means ± S.D, Sham group n=8, OVX group n=8, OVX+A group n=6, OVX+J group n=8,. Data were analyzed by one-way ANOVA test.