to be continue....網頁更新中....敬請期待

Gut microbiota plays an important role in human health and metabolism. Microbes produce variety of metabolites that can regulate host metabolism. The interactions between microbiota and the host has become important research topic. Short chain fatty acids (SCFA) are metabolites produced by gut microbes from the digestion of carbohydrate and/or cellulose. The epithetical cells in the gut use SCFA to maintain intestinal homeostasis. Vitamin B6, also known as pyridoxine, is an important coenzyme in heme and neurotransmitters production, as well as in carbohydrate and amino acid metabolism. In this study, C57BL/6 mice received B probiotic (Bp) and H probiotic (Hp), n=6/group for a period of time and then obesity was induced by a 60% high fat diet- (DIO, Research Diet) for ten weeks. Labeled ¹³C-glucose was used to trace SCFA pathway by gas chromatography-mass spectrometry. Quantitative analysis of vitamin B₆ was done by high performance liquid chromatography (HPLC). Feeding mice with a combination of Hp or Bp fomulas can increase the content of vitamin B6 and the richness of intestinal flora for improving health. In the future, these probiotics can be used for further developments of feed additives or health foods.

近年許多研究發現腸道內共生菌在人體吸收、代謝、整體健康中扮演重要角色。人類攝食不僅提供自身所需能量，也提供營養源給腸道中共存的菌叢。腸道菌群亦會透供應特定代謝產物給宿主，進一步調節宿主生理機能，因此腸道菌群產生之各種代謝產物與其宿主間之交互作用日益受重視。短鏈脂肪酸是腸道菌叢重要代謝產物。短鏈脂肪酸多由菌體將膳食纖維等醣類消化生成，這些短鏈脂肪酸有助於腸道黏膜之生成與功能。另外不同型態之維生素 B₆ 在生物體內參與多種反應，包括醣類、脂質、蛋白質代謝、紅血球生成及神經傳導等扮演關鍵輔酶的角色。本研究以 60% 高脂飲食(diet-induced obesity, DIO, Research Diet) 引起C57BL/6 雌性小鼠肥胖,分別給予 B 菌劑與 H 菌劑配方組合 (n=6/組log cells= 6.89)並利用穩定同位素碳 13 標定的碳源及質譜系統探討脂肪酸代謝路徑, 並利用 HPLC 分析小鼠各型態之維生素 B6 含量以探討益生菌對宿主營養代謝之影響。未來可以利用這些菌劑組合，進一步應用於飼料營養添加劑或保健食品之開發。

The present invention relates to a novel probiotic formula for increasing vitamin B6 contents in vivo, comprising live bacteria of a Clostridium butyricum or a Bacillus coagulans strain or the combination thereof. The probiotic formula of present invention can effectively enhance the activation and content of vitamin B6 in vivo and reduce the elimination of vitamin B6 in the body. Therefore, it can be used as a vitamin B6 promoter for the healthy people or patients with B6 deficiency.
本發明係提供一種可提升體內維生素B6 (尤其是活化型維生素B6)濃度的新穎益生菌配方，特徵在於包含丁酸梭菌 (clostridium butyricum)菌株或凝結芽孢桿菌 (bacillus coagulans)菌株之活菌，或其組合。本發明之益生菌配方能有效提升體內維生素B6之活化及利用，減少維生素B6的排除，可供做為健康者或缺乏B6疾病族群體內之維生素B6促進劑。

近年許多研究發現腸道中共生菌在人類的營養吸收與代謝中扮演十分重要的角色，人類攝取食物不只提供自己所需要的能量，同時也為其腸道中共存的菌叢提供營養來源。於此同時，腸道菌亦透過供應特定代謝產物給宿主、進一步調節宿主生理機能，因此腸道菌群所產出之各種代謝產物與其宿主間之相互影響更受重視。短鏈脂肪酸是腸道菌叢重要代謝產物，研究顯示這些短鏈脂肪酸多由腸道益生菌產生，由菌體將醣類與膳食纖維等消化生成，而結腸細胞會利用這些短鏈脂肪酸影響腸道黏膜之生成及功能。研究室學長姐先前研究曾發現特定益生元之菌劑組合會影響小鼠脂肪酸代謝、然而這些益生元與菌劑組合間、以及與宿主間的交互作用尚未被闡明。至於組合性益生菌在不同環境下如何影響不同碳源生成短鏈脂肪酸之路徑? 短鏈脂肪酸是否不同? 益生元供應的介入如何影響短鏈脂肪酸其生成利用? 腸道菌又會如何影響小鼠腸道的脂肪酸代謝，值得研究、也是我想探討的問題。本研究擬利用穩定同位素碳13 標定的碳源來追蹤宿主與菌脂肪酸代謝途徑以及其中的交互做用。將在體外試驗中探討不同腸道菌及益生元組合其脂肪酸產量之碳源利用途徑。選取特定益生元組合，探討在添加益生元情況下菌株生長與代謝之差異。從體外試驗結果組合成菌劑配方組合進一步進行動物實驗。體內試驗將以特殊菌組合及益生元配方菌劑組合餵食 C57BL/6J 小鼠，並同步追蹤其在正常與高脂飲食下腸道中利用葡萄糖等碳源之代謝流向。藉由體外菌組實驗比對小鼠腸道中的脂肪酸分佈，探討菌體本身或菌體介入影響宿主利用碳源之相關性。

專利獲證 單碳代謝為生物體內甲基的代謝過程，包含了葉酸循環、同半胱氨酸轉硫及轉甲基反應、甲硫胺酸及腺苷甲硫胺酸生合成、核苷酸合成等重要代謝路徑。 不同的研究暗示能量平衡可能與單碳代謝關係密切：先前研究發現肥胖者其血中甘胺酸的濃度較低。此外運動則會降低正常馬模型的血中甘胺酸和絲氨酸的濃度。有研究顯示限制卡路里在果蠅模型中會增加二磷酸腺苷和單磷酸腺苷的合成。在酵母菌模型中則會維持三磷酸腺苷的濃度。這些變化可能會造成體內單碳代謝的改變。本研究的目的在於探討長期餵食高脂飲食、長期有氧運動、長期阻力運動以及和長期限制卡路里對於體內單碳代謝之影響。

Background Bioactive food components are nonessential nutrients that are present in foods; many of them have been found to elicit the capacity to promote human health by modulation of one or more metabolic processes. The objective of this study was to examine the regulatory effects of specific bioactive food component supplementation on the protein abundance, gene expression and the activity of the key enzymes involved in adoMet homeostasis. Study design Study 1 was conducted to investigate the impacts of compound A supplementation on one-carbon kinetics in mice at young age. Study 2 was conducted to investigate the effects of parental compound A supplementation (during mating, gestation and lactation) on one-carbon metabolism in the offspring. In study 1, twelve male C57BL/6J mice aged 3 weeks were randomly assigned to receive either the control (CTL) or the compound A supplemented diet for 4 weeks. In Study 2, all parental mice received the compound A supplemented during mating, gestation and lactation. After weaning, the littermates continually received either the control diet or the compound A supplemented diet for an additional period of 4 weeks. Result. Compound A supplementation at a young age but not during gestation altered the hepatic and whole blood S-adenosylmethionine homeostasis. The isotopic tracer studies indicated that compound A supplementation tended to promote homocysteine remethylation fluxes in the tissues. Conclusion. The present study gives us new insights into the effects of compound A supplementation on homocysteine metabolism.

背景 先前文獻及本研究室先前研究顯示食品中有些活性物質會改變體內同半胱胺酸或硫-腺核?甲硫胺酸的含量。前者為心血管疾病的指標，後者為體內的甲基提供者，兩者皆為單碳代謝途徑中的關鍵代謝物，已知單碳代謝生化路徑的改變與許多人類病理狀態有關。本論文以動物模型探討不同的食品活性物質對於單碳代謝的影響。建立兩種動物模型，分析複合物A對S-腺基甲硫氨酸代謝途徑以及其相關代謝產物如S-腺核同半胱胺酸、同半胱胺酸及其相關代謝酵素活性、代謝酵素蛋白和基因表現量之影響。此外以同位素標定的方法去探討體內甲基代謝相關路徑分流的變化。

Background Bioactive food components are nonessential nutrients that are present in foods; many of them have been found to elicit the capacity to promote human health by modulation of one or more metabolic processes. The objective of this study was to examine the regulatory effects of specific bioactive food component supplementation on the protein abundance, gene expression and the activity of the key enzymes involved in adoMet homeostasis. Study design Study 1 was conducted to investigate the impacts of compound A supplementation on one-carbon kinetics in mice at young age. Study 2 was conducted to investigate the effects of parental compound A supplementation (during mating, gestation and lactation) on one-carbon metabolism in the offspring. In study 1, twelve male C57BL/6J mice aged 3 weeks were randomly assigned to receive either the control (CTL) or the compound A supplemented diet for 4 weeks. In Study 2, all parental mice received the compound A supplemented during mating, gestation and lactation. After weaning, the littermates continually received either the control diet or the compound A supplemented diet for an additional period of 4 weeks. Result. Compound A supplementation at a young age but not during gestation altered the hepatic and whole blood S-adenosylmethionine homeostasis. The isotopic tracer studies indicated that compound A supplementation tended to promote homocysteine remethylation fluxes in the tissues. Conclusion. The present study gives us new insights into the effects of compound A supplementation on homocysteine metabolism.

背景 先前文獻及本研究室先前研究顯示食品中有些活性物質會改變體內同半胱胺酸或硫-腺核?甲硫胺酸的含量。前者為心血管疾病的指標，後者為體內的甲基提供者，兩者皆為單碳代謝途徑中的關鍵代謝物，已知單碳代謝生化路徑的改變與許多人類病理狀態有關。本論文以動物模型探討不同的食品活性物質對於單碳代謝的影響。實驗設計 建立兩種動物模型，分析複合物A對S-腺?基甲硫氨酸代謝途徑以及其相關代謝產物如S-腺核?同半胱胺酸、同半胱胺酸及其相關代謝酵素活性、代謝酵素蛋白和基因表現量之影響。此外以同位素標定的方法去探討體內甲基代謝相關路徑分流的變化。實驗設計1為當代研究，探討早期餵食四週複合物A對當代老鼠代謝之影響。以複合物A混入飼料給予餵食四週後犧牲、取得各組織分析。實驗2為跨代研究，探討親代在交配、懷孕及哺乳期間補充上述複合物A對其子代代謝之影響。親代公母鼠在交配懷孕哺乳期間均攝食複合物A補充之飼料，子代斷奶後繼續餵食複合物A補充隻飼料四週進行分析。結果 在動物模型中，複合物A補充對當代及子代有不同影響。在當代鼠研究中,肝臟中腺?基甲硫氨酸增加,同時肝臟中腺?同半胱胺酸顯著性地下降。然而在子代鼠並未見到類似的情況。兩種動物模型中複合物A在不同組織中代謝路徑則較類似。複合物A會促進當代及子代肝臟、骨髓及大腦中同半胱胺酸再甲基化路徑。然而當代鼠主要顯現出較低腺?同半胱胺酸，而子代老鼠則在血漿中的同半胱胺酸有顯著減少現象。結論 當代及跨代餵食複合物A可透過改變肝臟及血液中的甲硫氨酸腺?基?移?及其他特定基因表現，進而改變相關代謝產物和代謝途徑。

to be continue....敬請期待

The concepts of ‘synergy” and “multifunctional” have driven the development of complex formulas in the global market of functional foods. The evident efficacy of these formulas could be achieved by a combination of multi-

ingredients from various food materials with health benefits. The objective of the present study was to design and investigate the efficacy of multi-ingredient and multifunctional health food formulas on improving vascular health. Six different multi-ingredient formulas containing bioactive compounds were designed and prepared from various food materials based on scientific evidence on vascular health promotion. Chemical analyses were performed on vitamins involved in vascular health. Hypercholesterolemia was induced by high cholesterol diet and in hamster model. Complex formulas were either supplemented in diet or fed by gastric gavage. Blood lipid profiles including cholesterol and triglyceride were examined. Liver lipid profiles and fecal lipid excretion were investigated. To investigate the in vivo homocysteine-lowering capacity of our multifunctional formulas, hyperhomocystemia was induced by diet in mice, followed by supplementation of the selected multi-

ingredients formula. Biochemical indices of homocysteine metabolism were investigated. The contents of vitamin E, B6, B12, folic acid and β-carotene were in low levels in all formulas, possibly due to the processing procedure. Administration of formula A, B and C did not alter homocysteine concentrations or other related metabolites. On the other hand, low dosage formula A as well as high dosage formulas B, I and J all significantly lowered serum triglyceride and total cholesterol in hamsters. The contents of hepatic lipids, triglyceride and cholesterol were decreased by above formulas with correlations observed between blood and hepatic lipid profiles. Formulas I, K and J increased excretion of fecal lipid and cholesterol excretion. Furthermore, high dosage formula J reduced hepatic total lipids and cholesterol. In addition, formula J significantly and dose-dependently reduced feeding efficiency, perirenal and epididymal adipose tissues in hypercholesterolemic animals. We suggested that above formulas which promoted the excretion of fecal lipids could simultaneously reduce accumulation of hepatic lipids and cholesterol as well as reduce abdominal body fat hence having potential on vascular health promotion.

『複合性』及『協同作用』的概念已是國際間保健食品開發的重點之ㄧ，藉由各種保健食材本身具有之生理活性，使用一種以上的材料並結合各成份不同的作用機制，可發揮更全面及顯著的保健功效。本研究選擇數種由天然植物取得之材料，以其萃取粉末混合搭配後製備成 A、B、C、I、K及J 等六種食品配方，並且以動物試驗探討這些配方對於調節血脂與心血管疾病相關之危險因子的功能性，以開發具有複合性特質的保健食品。

在高同半胱胺酸 (homocysteine, Hcy)的小鼠中，餵食配方A、B或C並無法顯著改善血中同半胱胺酸的濃度以及相關生化值。雖以各式天然蔬果為配方主要素材，維生素分析結果卻發現各配方中維生素E、B6、B12、葉酸及β-胡蘿蔔素等含量皆顯著偏低，可能流失於配方製程中，因此配方對於心血管疾病相關之危險因子-同半胱胺酸之影響有限。而在血脂調節的功能方面，低劑量的A配方與高劑量的B、I及J配方均會顯著降低高膽固醇倉鼠血清中三酸甘油酯和總膽固醇的含量，並且發現在這些配方劑量組別的肝臟脂質、三酸甘油酯或總膽固醇的含量也會同時減少，顯示血清脂質含量反應肝臟脂質狀況。此外，配方I、K與J皆會顯著增加糞便中脂質及膽固醇的排出，且與配方J呈現劑量效應 (dose-dependent)，因而可減少體內脂質堆積的機會。另外發現配方J的高劑量會明顯降低高膽固醇倉鼠體重的增加量，其食物利用率 (feeding efficiency) 與腎臟周邊及副睪的脂肪含量亦顯著減少並且與劑量呈現負相關。上述配方可以透過降低血清與肝臟脂肪或膽固醇、或增加脂質膽固醇的排出、減少食物利用率、減少體重的增加、減少腎周圍及睪丸周圍之脂肪堆積量，從而減少心血管疾病風險，因而提升血管健康，具有開發成為保健食品的潛力 (國科會計畫)。