本文關(guān)鍵詞： 骨質(zhì)疏松癥 脂肪干細(xì)胞 去卵巢小鼠 成骨分化　出處：《四川醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：骨質(zhì)疏松癥(Osteoporosis, OP)是一種系統(tǒng)性骨病,其發(fā)病機(jī)制是成骨細(xì)胞和破骨細(xì)胞在骨重塑過程中失衡,成骨細(xì)胞數(shù)量和活性降低,破骨細(xì)胞數(shù)量增加、功能活躍,導(dǎo)致骨吸收大于骨形成,造成骨質(zhì)疏松。大量研究證明脂肪干細(xì)胞(Adipose-derived stem cells,ASCs)具有向成骨細(xì)胞分化的潛能。骨質(zhì)疏松癥對脂肪干細(xì)胞生物學(xué)特性以及成骨分化的影響尚未得到完整闡釋。本實(shí)驗(yàn)通過對骨質(zhì)疏松癥小鼠和正常小鼠來源的兩組脂肪干細(xì)胞成骨能力的比較,探討骨質(zhì)疏松癥對脂肪干細(xì)胞成骨分化的影響,從而為骨質(zhì)疏松癥患者自體脂肪干細(xì)胞移植,修復(fù)骨缺損奠定基礎(chǔ)。方法：將雌性C57BL/6小鼠隨機(jī)分為實(shí)驗(yàn)組(去勢組)和對照組(假手術(shù)組)。分別獲取兩組小鼠腹股溝處脂肪組織來源的脂肪干細(xì)胞,體外純化擴(kuò)增至第3代。對細(xì)胞進(jìn)行成骨方向誘導(dǎo)后,使用茜素紅染色法檢測脂肪干細(xì)胞的成骨能力及組間區(qū)別；實(shí)時熒光定量PCR、免疫熒光染色以及Western Blot檢測脂肪干細(xì)胞成骨相關(guān)基因mRNA和成骨相關(guān)蛋白的表達(dá)。結(jié)果：茜素紅染色結(jié)果顯示,骨質(zhì)疏松來源的ASCs鈣鹽沉積明顯少于對照組,而real-time PCR和免疫熒光染色結(jié)果均指出,與對照組ASCs相比,實(shí)驗(yàn)組的ASCs細(xì)胞內(nèi)成骨相關(guān)基因mRNAs的轉(zhuǎn)錄以及相關(guān)蛋白的表達(dá)量均顯著降低。以上結(jié)果表明,與對照組相比,去勢后的小鼠來源的脂肪干細(xì)胞成骨向分化能力降低(P0.05)。結(jié)論：1.通過切除雌性小鼠雙側(cè)卵巢,成功構(gòu)建出骨質(zhì)疏松癥動物模型；2.來自去勢組的OP-ASCs較假手術(shù)組的ASCs增殖能力顯著降低；3.OP-ASCs成骨分化能力較正常ASCs顯著降低。
[Abstract]:Objective: Osteoporosissis (OPO) is a kind of systemic bone disease. Its pathogenesis is the imbalance of osteoblasts and osteoclasts in the process of bone remodeling, the decrease of the number and activity of osteoblasts, the increase of the number of osteoclasts and the active function of osteoclasts. Resulting in bone resorption greater than bone formation, A large number of studies have proved that adipose stem cells (Adipose-derived stem cells) have the potential to differentiate into osteoblasts. The effects of osteoporosis on the biological characteristics and osteogenic differentiation of adipose stem cells have not been fully explained. To compare the osteogenic ability of adipose stem cells between osteoporosis mice and normal mice, To investigate the effect of osteoporosis on the osteogenic differentiation of adipose stem cells, so that autologous adipose stem cells can be transplanted into the patients with osteoporosis. Methods: female C57BL / 6 mice were randomly divided into experimental group (castrated group) and control group (sham operation group). The osteogenic ability of adipose stem cells was detected by alizarin red staining method. Real-time fluorescence quantitative PCR, immunofluorescence staining and Western Blot were used to detect the expression of osteoblast related gene mRNA and osteoblast-associated protein in adipose stem cells. Results: the results of alizarin red staining showed that the deposition of calcium salt in ASCs from osteoporosis was significantly lower than that in control group. The results of real-time PCR and immunofluorescence staining showed that the transcription of osteoblast-associated gene mRNAs and the expression of related protein in ASCs cells in the experimental group were significantly lower than those in the control group, which indicated that compared with the control group, the expression of osteoblast-associated gene mRNAs in the experimental group was significantly lower than that in the control group. The osteogenic differentiation ability of adipose stem cells from ovariectomized mice decreased P0.050.Conclusion: 1. By removing the bilateral ovaries of female mice, OP-ASCs from ovariectomized group was significantly lower than that of sham operation group in ASCs proliferation. 3. Osteogenic differentiation ability of OP-ASCs was significantly lower than that of normal ASCs.
【學(xué)位授予單位】：四川醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R580

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