蛇床子素複方食品可延緩骨質流失

A study on ingestion of osthole formula to reduce bone loss

林雍原(Yung-Yuan Lin)

指導教授 ： 林淑容 ； 蔡敬民

中原大學/工學院/生物科技研究所/碩士(2014年)

https://doi.org/10.6840/cycu201400750

摘要

摘 要

本實驗的目的是欲探討本實驗室與生展生物科技公司共同設計之蛇床子素複方食品（OF）是否具有「有助於骨質保健」的功效? 實驗方法是根據衛生福利部於2013年02月05日公告修訂「健康食品之骨質保健功效評估方法」中的更年期動物模式試驗。動物選用10週大Wistar雌鼠40隻，並於正式實驗前先進行卵巢切除手術，以模擬婦女停經後之生理狀態。待2週讓開刀傷口恢復之後，將動物隨機（randomly）分配到5組，每組8隻，其中1組於分配後犧牲，供作為「起始組」；其餘之4組分別為「對照組」及攝取添加0.3%、0.6% 或1.2% 的含蛇床子素複方飼料之「實驗組」，動物飼養期為24週。
所有的動物於犧牲前將動物麻醉，使用雙能量X光吸收測定法（Dual-energy X-ray absorptiometry, DXA）測量其脊椎、股骨與脛骨之骨密度。飼養實驗期間記錄每隻動物每天的攝食量及每週之體重與身長。動物於犧牲時採血，供作血液中的造骨細胞（Osteoblasts）的活性指標「骨鈣素」與蝕骨細胞（Osteoclasts）的活性指標「第一型膠原蛋白之氮端胜肽素（NTx）」的測定；而於犧牲後取出腰脊骨與股骨，供進行micro-CT測定，觀察骨小樑數量、體積、厚度、間距及結構模型指數（SMI）之變化，以探討攝取試驗樣品達某一劑量是否能減緩切除卵巢後之骨質流失？
實驗結果顯示：攝食量、體重、軀幹長度、體長及腿長在各組間並沒有顯著的變化，DXA的測定結果顯示：去除卵巢的控制組之脊椎、股骨及脛骨之骨密度很明顯低於起始組（p<0.01），但3種不等劑量之蛇床子素複方組的這些骨骼之骨密度均顯著高於控制組的骨密度（p<0.05），且0.6%與1.2%劑量蛇床子素複方組的骨密測定值與起始組相比並無明顯差異（p>0.10），1.2%劑量組稍微高於0.3%劑量組（p<0.10)。骨小樑體積（BV/TV）、骨小樑厚度（Tb.Th）與骨小樑數量（Tb.N）的測定結果顯示：3種不同劑量蛇床子素複方組皆顯著高於控制組（p<0.05），而0.6%與1.2%劑量的蛇床子素複方組之骨小樑厚度則與起始組並無明顯差異（p>0.05），但骨小樑體積與骨小樑數量仍低於起始組（p<0.05）；而3種劑量蛇床子素複方組的骨小樑間距（Tb.Sp）則明顯小於控制組（p<0.05），表示骨骼中的空隙較小，但大於起始組（p<0.05）；3種劑量組間則無明顯差異（p>0.10）。從SMI的測定值可看出骨組織的型態，起始組的動物SMI接近0，表示骨骼較偏向板狀，而蛇床子素複方組的動物其SMI數值大於1，表示其骨骼型態偏向板柱狀，而控制組SMI接近於2，表示其骨骼更偏向柱狀。
本研究之血液骨生化指標分析顯示：動物血液中「骨鈣素」濃度於卵巢切除24週後明顯比起始值下降（p<0.05），而隨著攝取蛇床子素配方劑量的增加，動物的骨鈣素濃度明顯有比對照組增加的趨勢（p<0.05）；反之，「第一型膠原蛋白之氮端胜肽素」 （NTx）濃度會於卵巢切除24週後非常明顯上升（p<0.01），但攝取蛇床子素複方的3個劑量組之增加幅度都明顯比對照組低（p<0.05），然而3個劑量組間的NTx濃度彼此無差異（p>0.10）。
綜合以上分析結果顯示本實驗的一些檢測指標結果都印證攝取0.3%每日總食物攝取量之蛇床子素複方食品能減緩切除卵巢動物之骨質的流失或有利於骨質之修補。

關鍵字

骨密度 ； 骨質保健 ； 蛇床子素複方食品 ； 骨質流失

並列摘要

To investigate the physiological function of osthole formula (OF), which was designed by our laboratory and Syngen Biotech Co. on bone loss, this study followed the guidelines of health food assessment, which was recommended by the Ministry of Health and Welfare. To begin, the study employed 40 ten-week old female Wistar rats. All rats were ovariectomized as menopause animal models and kept for two weeks under care for wound healing. Then, the rats were randomly divided into five groups of eight rats each. The animals in one of the five groups were sacrificed before the feeding experiment as an initial group. The second group was fed a normal diet. Then the subsequent groups were fed with a normal diet as the control group or added with 0.3%, 0.6%, or 1.2% Osthole formula (OF) respectively as the treatment groups for 24 weeks.
The bone mineral densities (BMD, g/cm2) of lumbar vertebrae, femurs and tibia of each rat were measured by using dual-energy X-ray absorptiometry (DXA) under anesthesia before it was sacrificed. During the feeding experiment period, daily food consumptions and weekly body weights and lengths of each rat were measured and recorded. Blood was withdrawn for osteocalcin, a bone formation marker, and cross-link N-telopeptide (NTx), a bone resorption marker, determinations during sacrificing. Then, the lumbar vertebrae, femurs and tibia were also obtained after sacrifice for trabecular bone determinations, including numbers, volume, thickness, separation and structure model index by using microfocus computed tomography system (micro-CT). The experiment was designed to investigate if the ingestion of osthole formula OF would prevent or slow down the bone loss in overiectomized rats.
The results showed that feed intake, body weight, trunk length, body length and leg length were the same among the feeding groups. DXA measurements demonstrated that the bone densities of lumbar vertebrae, femurs and tibia in control group were very significantly lower than initial group (p<0.01). It also showed that those bone densities in all three OF groups were higher than the control group (p<0.05). Results revealed that the bone densities in 0.6% OF and 1.2% OF groups were almost same with the ones in initial group (p>0.10), however, the ones in 1.2% OF group was slightly higher than ones in 0.3% OF group (p<0.10).
Results of the trabecular bone determinations of lumbar vertebra, femurs neck and femurs distal by micro-CT showed that the trabecular bone volumes (BV/TV, %), thicknesses (Tb.Th, mm) and numbers (Tb.N, /mm) in all three OF groups were higher than the ones in control group (p<0.05), the trabecular thicknesses in 0.6% and 1.2% OF groups were almost same as the ones in initial group (p>0.05). But, the trabecular bone volumes and numbers from OF groups were still lower than the ones in initial group (p<0.05). The trabecular bone separations (Tb.Sp, mm) in all three OF groups were smaller than the ones in control group (p<0.05), indicating more compacted, but they were still larger than the ones in initial group (p<0.05). However, no difference in trabecular bone separation was found among these three OF groups (p>0.10). The Structure Model Index (SMI) is a measurement of the relative ratios of rod- and plate-like trabecular structures. The initial group had a relatively smaller SMI, approaching 0, indicating more plate-like structure. The control group had relatively larger SMI approaching 2 indicating more rod-like trabecular structure. The three OF groups had SMI values in between, but larger than 1, representing a mixture of plates and rods, but relatively more rod-like than plate-like structure.
The data of bone biochemical markers in serum demonstrated that osteocalcin, a bone formation marker, was significantly reduced (p<0.05) in control group compared with the initial group after ovariectomy for 24 weeks. The results also revealed increasing the doses of OF intake could enhance the levels of serum osteocalcin (p<0.05). However, the levels of NTx, a bone resorption marker, was very significantly promoted (p<0.01) in control group compared with the initial group after ovariectomy for 24 weeks. The results showed that ingestion of OF could decrease serum NTx levels (p<0.05), however, no difference could be observed among these three various dose OF groups (p>0.10).
In conclusion, all the result parameters demonstrated that the supplement of osthole formula (OF) with a dose of 0.3% daily total diet intake could reduce bone loss and restore the bone health in ovariectomized rats.

並列關鍵字

trabecular bone ； bone mineral density ； Bone loss ； osthole formula

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