- 學位論文
咖啡因可促進蝕骨細胞活性而降低骨質密度
Caffeine enhances osteoclast activities and reduces bone densities
摘要
本實驗主要藉由細胞實驗與動物實驗兩部分探討咖啡因對於骨骼前驅細胞分化、活性及成長期大白鼠骨質密度之影響。細胞實驗之細胞取自雄性ICR小鼠，首先以細胞毒性試驗測試本實驗擬使用的劑量是在對間葉幹細胞對咖啡因絕不會因滲透壓等而產生毒害範圍內。然後以高低二不同濃度之咖啡因處理骨骼前驅細胞，觀察咖啡因對於骨形成作用、脂肪細胞新生作用、蝕骨細胞新生作用及骨質回收作用之影響。動物實驗部分為期20週，所採用實驗動物為3週大雄性Wistar大白鼠50隻，隨機分成5組，分別餵食control diet、0.1% caffeine、0.2% caffeine、0.002% atorvastatin及0.2% caffeine + 7.0% lard + 0.5% cholesterol。進入實驗前先以雙能量X光測定儀測量其脊椎骨、股骨及脛骨的起始骨密度，並於進入實驗後10週與20週時再測其骨質密度。實驗結束時，測定其股骨與脛骨之鈣含量。 實驗結果顯示，以0.005mM與0.01mM caffeine處理間葉幹細胞，對於間葉幹細胞分化成造骨細胞並沒有明顯影響，但是於濃度0.01mM時會抑制其分化成脂肪細胞 (p<0.05)。而0.005mM與0.01mM caffeine會促使造血幹細胞分化成蝕骨細胞 (p<0.05)，並且促進蝕骨細胞活性而增加其骨質回收作用 (p<0.05)。另外，以0.01mM genistein處理間葉幹細胞，會促使間葉幹細胞分化成造骨細胞 (p<0.05)，並會抑制其分化成脂肪細胞 (p<0.05)，且其會稍微抑制造血幹細胞分化成蝕骨細胞與其骨回收作用 (p<0.10)。 動物實驗結果顯示，0.2% caffeine會使成長期大白鼠脊椎骨、股骨及脛骨骨質密度增加量皆比control組小 (p<0.05)，實驗亦發現0.2% caffeine組的股骨鈣含量略低於control組 (p<0.10) ，脛骨鈣含量也明顯低於control組 (p<0.05)。攝取0.1% caffeine動物之股骨增加量也呈現略低於control組 (p<0.10) 之現象；另外，0.2% caffeine + 7.0% lard + 0.5% cholesterol組之股骨密度增加量會略低於control組 (p<0.10)，而0.002% atorvastatin與control組無差異 (p>0.05)。 綜合實驗結果顯示caffeine會促進造血幹細胞分化成蝕骨細胞，並且增強其骨質回收作用，導致成長期大白鼠骨質密度之增加會比較緩慢；而植物性雌激素genistein會促進骨形成作用，並會抑制脂肪細胞新生，且其會稍微抑制蝕骨細胞的形成與其骨質回收作用；降血脂藥物atorvastatin於本實驗使用之劑量條件下，對成長中的骨骼系統並無明顯的影響。
並列摘要
The study was to investigate the effect of caffeine on the differentiation of bone progenitor cells, cell activities and bone mineral density (BMD) in rats by in vitro and in vivo studies. In the in vitro trial, cells were obtained from male ICR mice. At the beginning of the study, it was pre-tested to make sure the doses might be used were not toxic to the cells due to osmotic pressure. Then, the effect of caffeine on bone formation, osteoclastogenesis and adipogenesis were investigated. In animal study, fifty male Wister rats were assigned into 5 groups randomly, 10 each, and fed control , 0.1% caffeine, 0.2% caffeine, 0.002% atorvastatin or 0.2% caffeine + 7.0% lard +0.5% cholesterol diet respectively for 20 weeks. The BMD of lumbar vertebra, femur and tibia at 0, 10 and 20 weeks were determined by Dual-energy X-ray absorptiometry (DEXA). The calcium concentration of femur and tibia were also measured after animals were sacrificed. The results of the in vitro trials showed that 0.005 mM caffeine or 0.01mM caffeine in diet could not affect the mesenchymal stem cells (MSC) to osteoblasts (OB) (p>0.05), but could enhance the differentiation of hematopoietic stem cells (HSC) to osteoclasts (OC) and also their bone resorption (p<0.05). It was observed that 0.01mM caffeine could inhibit the differentiation of MSC to adipocytes (p<0.05). Genistein could increase the differentiation of MSC to OB (p<0.05), and inhibit the differentiation of MSC to adipocytes (p<0.05). The results demonstrated that genistein inhibited the differentiation of HSC to OC and the bone resorption slightly (p<0.10). Data in the animal trials showed that BMD of lumbar vertebra, femur and tibia were lower in 0.2% caffeine group than in control group (p<0.05). The calcium concentrations of tibia in 0.2% caffeine group was lower than in control group (p<0.05), and in femur was also lower slightly than in control group (p<0.10). It was noticed that BMD of femurs in 0.1% caffeine and 0.2% caffeine + 7.0% lard + 0.5% cholesterol group were lower slightly than in control group respectively (p<0.10). However, 0.002% atorvastatin didn’t affect BMD of lumbar vertebra, femur and tibia comparing with control group (p>0.05). Conclusion of the study is that caffeine can increase the differentiation of HSC to OC and bone resorption. Therefore, caffeine can decrease BMD of bones in growing rats. Genistein, a phytoestrogen, can increase bone formation and inhibit adipogenesis. It also inhibits osteoclastogenesis and bone resorption. But, atorvastatin which can reduce serum cholesterol doesn’t affect BMD in growing male rats.
參考文獻
延伸閱讀
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