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  • 學位論文

北蟲草廢棄米培養基之抗肥胖功效探討

Study of anti-obesity effects of Cordyceps militaris spent rice media

指導教授 : 劉嚞睿
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摘要


肥胖率在現代社會中逐漸攀升並且容易增加第二型糖尿病、心血管疾病以及非酒精性脂肪肝等疾病的患病機率,使肥胖現象儼然成為已開發社會的健康威脅之一。許多研究即以傳統中草藥為材料,測試其抗肥胖功能,企圖尋找出安全有效的抗肥胖藥物。眾多中草藥皆被發現具有抗肥胖功效,而北蟲草(Cordyceps militaris)便是其一。北蟲草可由人工培養在米培養基上,在北蟲草子實體收割後米培養基即被視為廢棄物並丟棄。然而,考慮到廢棄米培養基中含有北蟲草菌絲體,猜測其或許也具有些許藥用功效與利用價值。本論文之目的即在驗證此一說之可行性,以3T3-L1細胞與SD大鼠為試驗模型,測試北蟲草廢棄米培養基(Cordyceps militaris spent rice media, CMS)在抗肥胖方面的效果。由於蟲草素(Cordycepin)普遍被認為是北蟲草中的功效成分,故首先以高效液相層析(high performance liquid chromatography, HPLC)測定CMS中蟲草素含量,發現CMS的蟲草素含量約占其總重之0.4%,確認其含有功效成分。在細胞試驗結果中顯示北蟲草廢棄米培養基水萃物(Cordyceps militaris spent rice media water extract, CMSWE)可透過增加AMPK與Wnt10b訊號路徑相關蛋白的mRNA表現量而抑制C/EBPα與PPARγ的表現,從而阻礙3T3-L1前脂肪細胞的分化。此外,CMSWE亦可透過抑制脂肪合成與促進脂肪分解、β氧化相關基因表現而降低3T3-L1細胞中的脂肪累積。動物試驗結果進一步顯示餵食CMS的大鼠減輕由高脂飼料誘發的肥胖現象,具有較輕的體重、較少的脂肪量以及較低的血清三酸甘油酯與膽固醇含量,同時CMS亦可減輕由高脂飼料引起之肝脂肪變性程度。此外,由第三代定序技術分析大鼠腸道菌相結果指出,餵食CMS的大鼠腸道中Akkermansia muciniphila與Parabacteroides goldsteinii二個菌種的豐度有所增加,而前人研究曾指出此二菌種對宿主的代謝具有正面影響,可能在CMS減輕肥胖症狀的效果中具有一定的作用。上述試驗結果均顯示北蟲草廢棄米培養基具有抗肥胖的能力與潛在的應用價值。

並列摘要


Obesity is prevalent in modern countries and notorious for increasing the risk of others diseases, such as cardiovascular disease and type 2 diabetes. In order to find the safety and effective treatment against obesity, lots of traditional Chinese medicines were tested for their anti-obesity ability, and Cordyceps militaris, a kind of medical fungus, is a potential candidate for obesity alleviation. Cordyceps militaris can be cultured on rice media. The fruit bodies of Cordyceps militaris are medically and economically valuable, while the rice media is regarded as waste and will be discarded after the fruit bodies are harvested. However, consider that the spent rice media contains mycelium of Cordyceps militaris, it seems to be reasonable and possible that the spent rice media might has medical activity. Therefore, we performed in vivo and in vitro experiments to test the anti-obesity ability of Cordyceps militaris spent rice media (CMS), and tried to explore its potential value. Cordycepin is the major bioactive compound that contribute to the anti-obesity effect of Cordyceps militaris. We measured cordycepin content of CMS by high performance liquid chromatography (HPLC) and found that its content in rice media was about one third of that in the fruit body, indicating the possibility of its medical activity. For in vitro test, 3T3-L1 cells were used as an experiment model. The cell experiment results showed that water extract of CMS (CMSWE) strongly inhibited adipogenesis of 3T3-L1 preadipocytes by upregulating mRNA expression of AMPK and proteins that related to Wnt10b signaling pathway, and consequently inhibited mRNA expression of C/EBPα and PPARγ. CMSWE treatment also reduced lipogenesis in mature 3T3-L1 adipocytes by inhibiting lipogenesis and promoting lipolysis. The in vivo test further revealed that oral administration of CMS reduced body weight and adipose tissue weight in high fat diet-induced obese rats. Serum concentration of triacylglycerol and cholesterol were lower in CMS administrated group as well. Moreover, rats fed with high fat diet developed severe hepatic steatosis, and this condition was alleviated by CMS administration. Third generation sequencing revealed the abundance of Akkermansia muciniphila and Parabacteroides goldsteinii, species which had been reported are beneficial to host, were increased in CMS administrated group. Overall, these data suggest that Cordyceps militaris spent rice media has anti-obesity capacity and potential application value.

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