第四章 研究結果
第四節 降低體脂肪之動物實驗評估
一、功能性指標評估
(一) 體重變化、攝食量與食物利用率
在起始帄均體重無顯著差異 (p>0.05) 為原則之下,將大鼠隨機分為 8 組,
每組 8 隻。實驗中同時給予高油脂飲食與不同試驗物質,餵食 8 週後,進行犧 牲及各項分析。結果如表 4-5 所示,高油脂組 (HF) 之體重明顯 (p<0.05) 高於 正常飲食組 (NOR),餵食一倍以 RO 水發酵之紅麴山藥組 (RO-RMD 1X) 與一 倍以深層海水發酵之紅麴山藥組 (DSW-RMD 1X) 之體重均呈下降之趨勢 (p <
0.05),而 DSW-RMD 1X 又較 RO-RMD 1X 具有更良好的降低體重的效果。餵 食含甘露二醣不含咖啡因之降體脂肪咖啡的正控制組 (POS)、兩倍以深層海水發 酵 之 紅 麴 山 藥 組 (DSW-RMD 2X) 、 純 物 質 monascin 組 (MS) 與 純 物 質 ankaflavin 組 (AF) 均有顯著 (p < 0.05) 降低體重的效果。雖然 DSW-RMD 無 呈現劑量效應,但結果顯示餵食 DSW-RMD 2X 之體重有較好的下降趨勢。
研究結果如表 4-6 顯示,高油脂飲食組 (HF) 之攝食量較正常飲食組 (NOR) 低 (p < 0.05),這可能是因為高油脂飲食之單位熱量較高 (HF:4.17 kcal/g 與 NOR:3.34 kcal/g),動物體內之回饋帄衡機制使得食物之攝取量下降。餵食 含甘露二醣不含咖啡因之降體脂肪咖啡的正控制組 (POS)、純物質 monascin 組 (MS) 與純物質 ankaflavin 組 (AF) 之攝食量均顯著 (p<0.05) 的少於高油脂飲 食組 (HF),在此推測餵食 monascin 與 ankaflavin 可抑制食慾,降低食物的攝 取量。
食物利用率 [(體重變化 / 總攝食量) × 100%] 可視作相同重量飼料增加動 物體重之能力。此部分結果如表 4-6 顯示,高熱量飲食組 (HF) 會明顯 (p<0.05) 提高飼料效率,這可能是因為高油脂飲食的單位熱量較高所致。餵食一倍以 RO 水發酵之紅麴山藥 (RO-RMD 1X) 其飼料效率無顯著差異 (p<0.05),而餵食一 倍以深層海水發酵之紅麴山藥 (DSW-RMD 1X) 有顯著降低飼料效率之趨勢,且 餵食深層海水發酵之紅麴山藥具有劑量效應。餵食純物質 monascin 組 (MS) 與 ankaflavin 組 (AF) 其飼料效率均顯著 (p<0.05) 下降,因此推測餵食純物質 monascin 與 ankaflavin 除了抑制食慾降低攝食量之外,應另有調節降低體脂肪 之途徑。
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表 4-5 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠體重之影響
Table 4-5 Effect of various samples on body weight in male SD rat fed with high fat diet.
Groups Initial body weight (g)
Final body weight (g)
Weight gain (g) NOR 463.0 ± 33.9 b 579.0 ± 37.6 bc 116.0 ± 20.9 b
HF 430.1 ± 37.4 ab 601.4 ± 42.0 c 171.3 ± 18.0 de POS 439.0 ± 14.9 ab 580.6 ± 24.9 bc 141.6 ± 16.0 c RO-RMD 428.5 ± 28.1 ab 605.4 ± 30.2 c 176.9 ± 22.3 e DSW-RMD 1X 436.9 ± 18.1 ab 591.9 ± 23.3 bc 155.0 ± 12.6 cd DSW-RMD 2X 417.5 ± 12.0 ab 565.8 ± 27.4 b 148.3 ± 20.7 c
MS 410.3 ± 20.8 a 514.5 ± 23.1 a 104.3 ± 5.2 ab AF 413.8 ± 17.0 a 512.1 ± 26.9 a 97.4 ± 11.4 a
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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表 4-6 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠的總熱量攝取、攝食 量及食物利用率之影響
Table 4-6 Effect of various samples on food intake and feed efficiency in male SD rat fed with high fat diet.
Groups Calorie intake (kcal/8 weeks)
Food intake (g)
Feed efficiency (%)
NOR 5833.3 ± 125.9 d 1746.5 ± 37.7 d 6.8 ± 1.2 a HF 5962.7 ± 203.1 bc 1429.9 ± 48.7 bc 12.0 ± 1.4 ef POS 5404.7 ± 171.8 a 1296.1 ± 41.2 a 9.0 ± 3.6 bc RO-RMD 6106.1 ± 143.9 c 1464.3 ± 34.5 c 12.7 ± 1.5 f DSW-RMD 1X 5858.9 ± 175.1 b 1405.0 ± 42.0 b 10.8 ± 0.9 de DSW-RMD 2X 5956.4 ± 176.4 bc 1428.4 ± 42.3 bc 9.8 ± 1.4 cd
MS 5262.5 ± 264.0 a 1262.0 ± 63.3 a 7.9 ± 0.6 ab AF 5447.3 ± 233.1 a 1306.3 ± 55.9 a 6.3 ± 1.2 a
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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(二) 總體脂肪重量與體脂率
不同試驗物質對高油脂飲食之雄性 SD 大鼠總體脂肪重量與體脂率之影 響,如表 4-7 所示。在總體脂肪量方面,DSW-RMD 1X 雖較 RO-RMD 1X 組 無顯著差異,但具有降低總體脂肪量之效果。DSW-RMD 2X、MS 與 AF 三組 均具有顯著降低體脂肪量之效果,即表示深層海水發酵之紅麴山藥具有降低體脂 肪之效果,而 monascin 與 ankaflavin 均為降低體脂肪之功效成分。
體脂率 [(脂肪量 / 體重) × 100%] ,是指總體重中,脂肪組織所佔的比率。
各試驗物質組之體脂率如表 4-7 所示,DSW-RMD 1X 較 RO-RMD 1X 組具有 顯著降低體脂率效果。雖 DSW-RMD 2X 與 DSW-RMD 1X 不具有劑量效應,
但有降低體脂率之趨勢。MS 與 AF 組均可顯著降低體脂率 (p<0.05)。由上述 結果表示,餵食深層海水發酵之紅麴山藥一倍與二倍劑量均可顯著降低體脂肪之 生成,一倍劑量可顯著降低體脂率,而二倍劑量僅具降低之趨勢。而以深層海水 發酵之紅麴山藥中之純物質 monascin 與 ankaflavin 均可抑制體脂肪增加與體 脂率的上升。
(三) 腎周圍體脂肪重量與腎周圍體脂率
不同試驗物質對高油脂飲食之雄性 SD 大鼠腎周圍體脂肪與體脂率之響,
如表 4-8。不論在腎周圍脂肪重量或腎周圍體脂率方面, DSW-RMD 1X 均可顯 著 抑 制 體 脂 肪 生 成 與 體 脂 率 的 上 升 , 且 其 效 果 都 較 RO-RMD 1X 更 好 (p<0.05)。雖然,DSW-RMD 不具有劑量效應,但 DSW-RMD 2X 具有降低之趨 勢 。 MS 與 AF 組 均 可 顯 著 抑 制 腎 周 圍 體 脂 肪 的 生 成 與 體 脂 率 的 上 升 (p<0.05)。綜合上述結果得知,以深層海水發酵之紅麴山藥可顯著的抑制腎周圍 體脂肪的生成與體脂率的上升,而 monascin 與 ankaflavin 則是功效成分。
(四) 副睪周圍體脂肪重量與副睪周圍體脂率
不同試驗物質對高油脂飲食之雄性 SD 大鼠副睪周圍體脂肪與體脂率之 響。如表 4-9,不論在副睪周圍脂肪重量或副睪周圍體脂率方面, DSW-RMD 1X 較 RO-RMD 1X 可抑制體脂肪生成與體脂率的上升之趨勢。雖然,DSW-RMD 不具有劑量效應,但 DSW-RMD 2X 具有降低之趨勢。MS 與 AF 組均可顯著 抑制副睪周圍體脂肪的生成與體脂率的上升 (p<0.05)。綜合上述結果得知,以深 層海水發酵之紅麴山藥可顯著的抑制副睪周圍體脂肪的生成與體脂率的上升,而 monascin 與 ankaflavin 則是功效成分。
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表 4-7 不同試驗物質對高油脂飲食之雄性 SD 大鼠總體脂肪重量與體脂率之 影響
Table 4-7 Effect of various samples on total fat pads weight and body fat pads percentage in male SD rat fed with high fat diet.
Groups Total fat weight (g) Body fat ratio (%) NOR 16.8 ± 2.6 a 3.2 ± 0.5 a
HF 27.6 ± 3.5 c 5.0 ± 0.9 c POS (mannobiose) 23.7 ± 4.0 bc 3.7 ± 0.6 ab
RO-RMD 1X 25.4 ± 7.0 bc 4.8 ± 1.1 c DSW-RMD 1X 22.8 ± 4.1 b 3.8 ± 0.7 ab DSW-RMD 2X 18.9 ± 2.2 a 4.2 ± 1.3 bc MS 16.5 ± 2.8 a 3.3 ± 0.5 ab AF 17.5 ± 2.5 a 3.6 ± 0.4 ab
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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表 4-8 不同試驗物質對高油脂飲食之雄性 SD 大鼠腎周圍體脂肪與體脂率之 影響
Table 4-8 Effect of various samples on perirenal fat pads weight and perirenal fat pads percentage in male SD rat fed with high fat diet.
Groups Perirenal fat pads weight (g) Perirenal fat ratio (%) NOR 10.4 ± 1.4 ab 1.5 ± 0.2 a
HF 17.0 ± 3.9 d 2.8 ± 0.7 d POS (mannobiose) 14.3 ± 1.7 cd 2.3 ± 0.4 bc RO-RMD 1X 14.9 ± 5.4 cd 2.5 ± 0.9 cd DSW-RMD 1X 12.8 ± 2.7 bc 2.1 ± 0.5 bc DSW-RMD 2X 10.7 ± 1.5 ab 1.9 ± 0.3 ab MS 9.4 ± 1.6 a 1.9 ± 0.3 ab AF 9.9 ± 1.4 ab 1.8 ± 0.3 ab
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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表 4-9 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠副睪周圍體脂肪與體 脂率之影響
Table 4-9 Effect of various samples on epididymal fat pads weight and epididymal fat pads percentage in male SD rat fed with high fat diet.
Groups Epididymal fat pads weight (g) Epididymal fat ratio (%) NOR 8.3 ± 1.5 ab 1.5 ± 0.3 a
HF 13.0 ± 1.5 d 2.2 ± 0.2 d POS (mannobiose) 10.0 ± 1.5 c 1.7 ± 0.26 bc
RO-RMD 1X 10.5 ± 1.9 c 1.8 ± 0.3 c DSW-RMD 1X 9.8 ± 1.3 bc 1.6 ± 0.2 abc DSW-RMD 2X 8.5 ± 1.2 ab 1.5 ± 0.2 ab
MS 6.9 ± 1.2 a 1.4 ± 0.2 a AF 6.9 ± 1.4 a 1.4 ± 0.2 a
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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(五) 腎周圍與副睪周圍脂肪細胞截面積
由表 4-10 結果顯示,DSW-RMD 1X 組較 RO-RMD 1X 組可顯著降低腎周 圍與副睪周圍脂肪細胞的截面積 (p<0.05),則表示深層海水發酵之紅麴山藥較 RO 水發酵之紅麴山藥可更顯著降低腎周圍與副睪周圍脂肪細胞的截面積,且其 功效性成分為 monascin 與 ankaflavin。在此推測,深層海水發酵之紅麴山藥可 能具有抑制脂肪細胞增生或脂肪細胞體積增大之效果。
二、安全性指標評估
(一) 血清脂質之影響
總膽固醇含量與三酸甘油酯、高密度脂蛋白膽固醇及低密度脂蛋白膽固醇含 量息息相關。
不同試驗物質對高油脂飲食之雄性 SD 大鼠血清脂質之影響,結果如表 4-11 所示。DSW-RMD 1X 組與 RO-RMD 1X 均無顯著降低血清中總膽固醇之 效用,而深層海水發酵之紅麴山藥無劑量效應。而 MS 與 AF 組血清中總膽固 醇含量也與 HF 組無顯著差異。DSW-RMD 1X 較 RO-RMD 1X 組可顯著降低 血清中三酸甘油酯之含量 (p<0.05)。而給予深層海水發酵紅麴山藥之二倍劑量雖 無劑量效應,但其中的 monascin 與 ankaflavin 均具有顯著降低三酸甘油酯之效 果 (p<0.05)。
在高密度脂蛋白膽固醇含量方面,一倍劑量的深層海水發酵之紅麴山藥與 RO 水發酵之紅麴山藥均具有提高高密度脂白膽固醇與降低高密度脂蛋白的效 用,且深層海水發酵之紅麴山藥對於提高高密度脂蛋白膽固醇之效果較 RO 水 發酵之紅麴山藥好。而 MS 與 AF 二組均可減少血清中低密度脂蛋白膽固醇的 含量,卻無提高高密度脂蛋白膽固醇之效果,由此證實 monascin 與 ankaflavin 可能是降低低密度脂蛋白膽固醇之功效成分。
(二) 血糖含量之影響
Barnard 等人以高油脂飲食或低脂質與醣類的複合飲食進行動物實驗,試驗
2 年後,結果顯示,大鼠已具有肥胖、高血壓、高血胰島素、高血三酸甘油酯等 病癥 (Barnard et al. 1993)。Barnard 延續先前的試驗,在 1998 年提出造成胰島 素抗性與高胰島素症的原因是由飲食所以起的,並非肥胖。而胰島素與醣類代謝 相關,且高血胰島素可能是導致其他代謝症候群病症之因子。高油脂飲食不同試驗物質對高油脂飲食之雄性 SD 大鼠血糖之影響,結果 如表 4-12 所示。不論是深層海水發酵之紅麴山藥或 RO 水發酵之紅麴山藥均
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表 4-10 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠脂肪細胞截面積之 影響
Table 4-10 Effect of various samples on cell cross-sectional area of adipocyte in male SD rat fed with high fat diet.
Groups Cell cross-sectional area of perirenal (μm²)
Cell cross-sectional area of epididymal (μm²) NOR 7677.0 ± 5254.6 b 5922.4 ± 2413.8 b
HF 11746.2 ± 5954.8 c 8690.6 ± 3495.6 c POS (mannobiose) 5521.3 ± 3947.3 b 619.4 ± 179.5 a
RO-RMD 1X 5477.7 ± 1653.2 b 5485.9 ± 2435.0 b DSW-RMD 1X 872.8 ± 500.8 a 596.8 ±184.2 a DSW-RMD 2X 993.8 ± 282.5 a 465.6 ± 287.5 a MS 676.5 ± 317.5 a 1130.4 ± 514.2 a AF 353.0 ± 192.5 a 1150.8 ± 384.7 a
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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表 4-11 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠血清脂質之影響 Table 4-11 Effect of various samples on serum lipidic parameters in male SD rat fed with high fat diet.
Groups TC (mg/dL)
TG (mg/dL)
HDL-C (mg/dL)
LDL-C (mg/dL) NOR 63.1 ± 9.3 a 62.6 ± 10.3 abc 28.3 ± 1.9 bc 24.2 ± 6.2 ab
HF 65.1 ± 8.6 a 94.5 ± 17.5 d 22.6 ± 2.4 a 31.5 ± 7.9 b POS
(mannobiose) 62.3 ± 6.6 a 63.1 ± 94.5 abc 26.0 ± 2.9 b 23.3 ± 5.5 ab RO-RMD 1X 63.5 ± 6.9 a 69.8 ± 8.4 c 27.3 ± 3.6 bc 27.2 ± 7.3 ab DSW-RMD 1X 64.6 ± 7.9 a 52.8 ± 8.5 a 28.8 ± 1.6 c 25.2 ± 6.6 ab DSW-RMD 2X 62.9 ± 2.3 a 56.0 ± 52.8 ab 26.3 ± 2.0 bc 22.0 ± 6.6 a
MS 67.0 ± 16.1 a 67.1 ± 14.5 bc 22.5 ± 3.5 a 24.5 ± 13.2 ab AF 60.2 ± 6.8 a 60.4 ± 4.6 abc 22.2 ± 1.1 a 18.6 ± 5.4 a
NOR, normal diet (3.34 kcal/g); HF, high-fat diet (4.17 kcal/g); RO-RMD 1X, RO-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 1X, DSW-RMD powder (1X, 27.81 mg/day 100 g bw) and high-fat diet; DSW-RMD 2X, DSW-RMD powder (2X, 55.62 mg/day 100 g bw) and high-fat diet; POS, containing mannose disaccharide (mannobiose) of coffee powder without caffeine (1X, 500.57 mg/day 100 g bw) and high-fat diet; MS, monascin powder (2X, 0.55 mg/day 100 g bw) and high-fat diet; AF, ankaflavin powder (2X, 0.08 mg/day 100 g bw) and high-fat diet; Data are presented as means ± SD (n=8).
Mean values within each column with different superscripts are significantly different (p<0.05)
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表 4-12 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠血糖含量之影響 Table 4-12 Effect of various samples on serum glucose in male SD rat fed with high fat diet.
Groups Glucose (mg/dL) NOR 174.3 ± 47.3 a
HF 251.6 ± 9.8 b POS (mannobiose) 184.5 ± 26.1 a
RO-RMD 1X 196.8 ± 18.5 a DSW-RMD 1X 183.5 ± 27.3 a
RO-RMD 1X 196.8 ± 18.5 a DSW-RMD 1X 183.5 ± 27.3 a