卵巢摘除誘發骨質疏鬆的小鼠模式之構築

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Please use this identifier to cite or link to this item: https://ir.cnu.edu.tw/handle/310902800/28956

Title:	卵巢摘除誘發骨質疏鬆的小鼠模式之構築 Construction of the mouse model with ovariectomy-induced osteoporosis
Authors:	羅彥鈞
Contributors:	生物科技系李國榮
Keywords:	微電腦斷層掃描成骨細胞雌激素骨質疏鬆症 Osteoblast Estrogen Osteoporosis Micro-CT
Date:	2015
Issue Date:	2015-10-21 17:06:13 (UTC+8)
Abstract:	在當今高齡國人好發疾病中，骨質疏鬆症一直以來對患者和其家庭造成健康和經濟上不小的負擔。骨質疏鬆症於檢查上的特徵為患者的骨質減少且變得脆弱，最終導致骨折的風險提高。骨質疏鬆症的患者多為女性，是由於更年期的到來伴隨著停經下，雌激素的停止分泌使的骨質開始逐漸流失，若未處理會有不小的機率形成骨質疏鬆症的發病。在治療手段中，藥物治療為最常見之方法，但因患者長期治療的需求會有副作用的問題出現，找尋其他的替代療法為未來研究趨勢。本研究使用切除卵巢的成年雌性小鼠來模擬停經後婦女的骨質疏鬆發病，以手術切除卵巢一段時間後觀察發病情形，於犧牲後收其脛骨，使用微電腦斷層掃描比較微結構的變化並以組織切片染色驗證微結構變化來自於細胞表現，針對以上兩點分析比較正常與發病者的差別做為日後研究之基礎。材料與方法本研究使用9週齡雌性C57BL/6小鼠32隻，平均體重為20克。小鼠飼養在符合標準的環境下。小鼠隨機分為2組: (a) 卵巢切除(ovariectomized, OVX) 組( n=16) 以及 (b) 對照組(Control) 組(n=16)。以手術切除卵巢模擬停經後婦女骨質疏鬆發病條件，實驗進行為期20週，於10週時兩組各犧牲6隻；20週時各組犧牲10隻，收取雙邊脛骨針對其微結構和組織學進行分析。結果兩組小鼠脛骨於微電腦斷層掃描分析下，在衡量骨質疏鬆症的數個參數: 骨組織/體積比(BV/TV) 、骨小樑厚度(Tb.Th) 、骨小樑分數量(Tb.N) 、骨小樑分離度(Tb.Sp) 相比較均有顯著差異，發病組數據表現達到骨質疏鬆症發病之標準；在組織學蘇木素與伊紅( hematoxylin and eosin, HE)與末端脫氧核?酸轉移?脫氧尿?三磷酸切口末端標記(Terminal deoxynucleotidyl transferase dUTP nick end labeling, TUNEL)染色的觀察，OVX組脛骨生長板以下之幹?端區域內有骨小樑損失和脂肪細胞堆積與顯著多於對照組之細胞凋亡訊號，驗證雌激素缺乏下誘發骨細胞凋亡數量增加與組織的變化。結論本研究以卵巢切除小鼠建立之仿停經後骨質疏鬆症發病之動物模型，微電腦斷層掃描觀察其影像與骨小樑參數變化；組織學上則著重於骨質流失與骨、脂肪細胞的堆積與細胞凋亡共三項。結果微電腦斷層掃描造影和骨小樑參數反映了骨質流失；組織切片染色上可見大量脂肪細胞堆積、骨小樑損失與骨細胞凋亡訊號，皆符合停經後骨質疏鬆症應有病徵，完成該疾病動物模型。 Osteoporosis is a health burden for patients and their families in our country. Patients with osteoporosis have osteopenia, it lead bone to become fragile and increase the risk of bone fracture. Most patients with osteoporosis is elderly females when they were postmenopausal estrogen deficiency. Currently, pharmaceutical therapies is the most common method for treatment and the side effects perlex patients during long-term treatment. Therefore, alternative therapies are necessary for preventing the patients with osteoporosis from deterioration. In this study, we will establish ovariectomized mice model to simulate of the postmenopausal osteoporosis for the further studies.The present study used 32 female C57BL/6 mice, 9 weeks of age, average weight 20g. Mice were equally divided into two groups; ovariectomized group(OVX) and control group. Tibias of OVX and control mice were harvested respectively after ten weeks and twenty weeks feeding. Micro-computed tomography (CT) analysis, hematoxylin and eosin(HE) staining and terminal deoxynucleotidyl transferase dUTP nick end labling (TUNEL) assay were performed to verify the microstructure changes in tibia.Micro-CT analysis revealed that bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and trabecular spacing (Tb.Sp) have significant difference between these two groups. Compared with control group, BV/TV, Tb.Th and Tb.N of OVX group revealed decease and Tb.Sp revealed increase. In HE staining, trabecular bone loss and adipocyte embolisms shown in OVX group but not in control group. Furthermore, TUNEL assay revealed that OVX group have more significantly apoptotic signal than the control group. Interestingly, the apoptotic signals either from OVX group or control group were major located in bone marrow cells not in trabecular bone and osteoblast of tibia metaphysis. Taken together, these results demonstrated that OVX mice constructed in our laboratory shown similar pathology of postmenopausal osteoporosis. These mice may provide a useful model to find a potential therapeutic approaches.
Relation:	網際網路公開：2020-09-10，學年度：103，53頁
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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