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Title: 臺灣天仙果萃取物對人類腸道上皮細胞株Caco-2鈣離子運輸與糖尿病去卵巢雌性小鼠骨質疏鬆之影響
Effects of Ficus formosana extract on calcium transport into human intestinal epithelial Caco-2 cells and osteoporosis in diabetic ovariectomized female mice
Authors: 沈賜川
Shen, Szu-Chuan
Wu, Swi-Bea
Ting, Yu-Wen
Tien, Ping-Yu
Keywords: 骨質疏鬆症
Ficus formosana
Caco-2 cells
diabetic ovariectomized female mice
calcium transportation
Issue Date: 2020
Abstract: 隨著人口急遽老化,骨質疏鬆症的盛行率也正持續攀升。骨質疏鬆症為臨床常見疾病,不僅會導致骨質流失最終造成患者骨質密度減弱,也會增加骨折的機率。臺灣天仙果 (Ficus formosana Maxim.) 為桑科榕屬植物,是民間普遍使用之食補藥材,具有促進骨骼生長及活血化瘀等功效。本研究利用人類大腸癌細胞株Caco-2細胞transwell seeding單層膜模式評估天仙果莖部熱水萃取物對腸道細胞促鈣轉運之效果,並且以糖尿病去卵巢小鼠動物模式評估其延緩骨質疏鬆之效果及可能機制。首先利用MTT assay測試臺灣天仙果莖部熱水萃取物之毒性,再以Caco-2細胞模擬腸道鈣離子轉運的細胞模式,於transwell上室同時加入鈣離子與天仙果萃取物後,分別在不同時間點抽取下室液,並以 Arsenazo Ⅲ為呈色劑利用分光光譜法測定鈣離子濃度。MTT細胞存活率測試結果顯示,天仙果萃取物於濃度25-500 ppm時對Caco-2細胞並無毒性產生。添加高低濃度(250-50 ppm)的天仙果莖部熱水萃取物處理,可明顯提升Caco-2腸道細胞的鈣離子轉運能力(p<0.05)及加腸道鈣轉運相關蛋白質Claudin 2、Claudin 12、TRPV6、PMCA1和CaBP-9k表現量(p<0.05),且也增加與維生素D調控鈣離子途徑相關蛋白VDR之表現與抑制CYP24A1降解1,25(OH)2D3的能力。在去卵巢之糖尿病雌性小鼠餵食含有高劑量天仙果莖部熱水萃取物之飼料6週後,其血清骨鈣素(Osteocalcin)顯著增加,而骨膠原蛋白碳末端肽鏈(C-telopeptide of type I collagen CTX-1)則顯著降低(p<0.05);另一方面,雖然骨密度(Bone mineral density)各組間無顯著差異,但是臺灣天仙果莖部熱水萃取物處理組之骨小樑的厚度與數目皆較去卵巢糖尿病小鼠為高,顯示具有延緩骨質酥鬆之效果。本研究結果可作為評估臺灣天仙果植株莖部開發骨骼保健膳食補充劑或食品時之參考。
Increasing calcium absorption is the most important and well known to prevent and cure osteoporosis in human been. Ficus formosana (FF), belongs to Moraceac Ficus genus, is a perennial evergreen shrub. The Ficus formosana has been used as folk medicine to cure osteoporosis for decades due to its ability on enhancing human bone growth and adjust menstruation ect. This study evaluates the effects of Ficus formosana roots hot water extract (FFE) on calcium absorption in human intestinal epithelial Caco-2 cells and anti-osteoporosis in diabetic ovariectomized female mice. The cell viabilities of Caco-2 cells were measured by MTT assay. The apical side of the transwell insert was used to assess the effect of FFE on calcium transport across Caco-2 monolayers. HBSS buffer was collected from the basolateral side and analyzed by Arsenazo Ⅲ to measure calcium ion concentration. The results show that FFE at the tested ranges of 25-250 ppm exhibited no cellular cytotoxicity in Caco-2 cells. The treatment of FFE revealed a significantly increase the total calcium transported across Caco-2 monolayers. Western blot analysis shows FFE increased Ca2+ absorption-related proteins includes Claudin 2, Claudin 12, TRPV6, PMCA1b and Calbindin-D(9K) in the Caco-2 cells (p<0.05).Also, FFE increased Vitamin D-inducible calcium transport related proteins VDR, and inhibited CYP24A1. In addition, treatment of FFE for 6 weeks significantly increased serum osteocalcin, whereas decrease serum C-terminal telopeptides type I collagen compared with diabetic ovariectomized female mice. Moreover, FFE increased trabecular thickness and numbers in diabetic ovariectomized female mice. The above observations suggest that FFE possesses potential as a material to develop nutraceuticals or dietary supplements on improving bone health.
Other Identifiers: G060751010S
Appears in Collections:學位論文

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