滋陰中藥對骨母細胞增生和礦化之影響

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Title:	滋陰中藥對骨母細胞增生和礦化之影響 The Effects of Nourishing Yin Chinese Herbal Medicine on Osteoblast Differentiation and Mineralization
Authors:	許少維 Hsu, Shew-Wei
Contributors:	許秀蘊
Keywords:	骨母細胞；女貞子；旱蓮草；黃花蜜菜；骨生成作用；礦化作用 osteoblast；nourishing yin herbal；bone formation；mineralization
Date:	2015-07-15
Issue Date:	2020-08-28 09:38:19 (UTC+8)
Abstract:	人體在進入更年期後，骨骼組織會因為其中的骨生成與骨吸收兩種作用的失衡而導致流失，引發骨質疏鬆症。同時全球的老年化人口比例逐年上升、經濟發展與醫療保健制度的改善，致使國人平均壽命逐年提高，使得老年化疾病比例持續上升，由於近年來從豆科與菊科植物中發現具預防骨質流失、增加骨密度的植物雌激素，且從古籍的中醫理論裏發現，藉由調補肝腎( 中醫臟腑 )可促使骨骼組織的生長，因此本研究欲確立以中草藥治療骨質疏鬆症的研究方向。本研究選用出生二至三天的ICR新生小鼠之骨母細胞作為實驗模式。以不同濃度之女貞子、旱蓮草和黃花蜜菜的中藥萃取液培養不同的天數，確定其是否會造成細胞毒性，接著以Alkaline phosphatase、Collagen type 1、Osteocalcin、 Osteoprotegerin及Receptor activator of nuclear factor kappa-B ligand等五種骨母細胞分化、礦化相關指標判斷其增生或礦化效果及其mRNA、蛋白質表現量的變化，最後證實滋陰中藥對骨母細胞分化、礦化的相關機制為何。實驗發現以滋陰中草藥萃取液在0.5 ~ 2.5 mg / mL的濃度下培養骨母細胞並無產生毒性，且不同種類都具有增生活性但各有差異；同時以RT-PCR、Western blot和ALP活性測試的結果，大部分滋陰中藥對骨母細胞之分化有一定程度的促進效果；此外，Col-1的mRNA和蛋白質大量表現，證明滋陰中藥對骨母細胞的骨基質合成能力有顯著的提升；而OPG、RANKL的mRNA和蛋白質表現量變化，證明部分滋陰中藥所含的活性天然成分在骨母細胞分化作用的期間，同時刺激並調控破骨細胞之生長因子與訊號途徑，以抑制其細胞的成熟、生長，而達到相對的骨母細胞增生；而關於滋陰中藥對骨母細胞中鈣堆積的變化，由OCN的mRNA、蛋白質表現和鈣堆積染色分析可得知，各組萃取液對細胞的鈣堆積在短時間下皆無特殊變化，除了女貞子組有緩慢得上升。綜觀本研究的實驗結果，滋陰中藥雖對骨母細胞的礦化能力無顯著的促進或抑制之作用，這牽涉到藥材中的天然活性成分，其含量因製備方式不同而產生的差異；而滋陰中藥仍對骨母細胞之分化作用及骨基質合成能力有促進及提升的作用，這表示滋陰中藥可作為特定飲食習慣的消費者在產生骨骼相關疾病一個新的替代療法，且在開發未來對骨質疏鬆症的預防及治療上具有一定的潛力。 Osteoblast, originated from mesenchymal stem cells ( MSCs ), is involved in the process for bone formation. Osteoblast, together with osteoclast, plays an important roles in the regulation of bone remodeling, which is called bone formation and bone resorption, respectively. Bone formation and resorption is a balanced and continuous process, which results in osteoporosis. Osteoporosis, a bone pathology related disease characterized by low bone mass and micro-archictectural deterioration of bone tissues, leads to enhanced bone fragility and consequent increase in fracture risk that results in fractures with minimal trauma. Ligustrum lucidum Ait., Eclipta prostrata L. and Wedelia calendulacea (L.)Less., are Chinese medicines used as “ Nourishing Yin herbs “, treat ailments such as menopausal problems, blurred vision, tinnitus, palpitations and insomnia, as well as to alleviate age-related symptoms. In this study, aimed to investigate the effects of the three herbs on differentiation of osteoblast by evaluating the osteoblast differentiation related markers ( including ALP, Col-1, OCN, OPG and RANKL ), after treating with herbs water extracts. The results showed that three herbs signiﬁcantly promote cell viability, whether alone or mixed treatment. Results of RT-PCR showed that relative expression of all markers except for OCN was up-regulated at the early differentiation stage, but down-regulated on the lately differentiation stage. Compared to control mRNA expression, the relative expression of OCN is not signiﬁcantly increase after treatment with three herbs water extracts. In conclusion, Nourishing Yin herbs signiﬁcantly promoted cell differentiation, but no effect on the bone mineralization. In terms of cell culture, each herbs promoted cell proliferation, and inhibited osteoclast growth, but there are no effect on the mineralization of osteoblast.
Description:	碩士指導教授:許秀蘊委員:李仁愛委員:孫瑞昇
Data Type:	thesis
Appears in Collections:	[School of Pharmacy] Dissertation/Thesis

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