骨质疏松小鼠脂肪干细胞成骨能力的研究

发布时间：2018-03-01 00:05

本文关键词： 骨质疏松症脂肪干细胞去卵巢小鼠成骨分化　出处：《四川医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：骨质疏松症(Osteoporosis, OP)是一种系统性骨病,其发病机制是成骨细胞和破骨细胞在骨重塑过程中失衡,成骨细胞数量和活性降低,破骨细胞数量增加、功能活跃,导致骨吸收大于骨形成,造成骨质疏松。大量研究证明脂肪干细胞(Adipose-derived stem cells,ASCs)具有向成骨细胞分化的潜能。骨质疏松症对脂肪干细胞生物学特性以及成骨分化的影响尚未得到完整阐释。本实验通过对骨质疏松症小鼠和正常小鼠来源的两组脂肪干细胞成骨能力的比较,探讨骨质疏松症对脂肪干细胞成骨分化的影响,从而为骨质疏松症患者自体脂肪干细胞移植,修复骨缺损奠定基础。方法：将雌性C57BL/6小鼠随机分为实验组(去势组)和对照组(假手术组)。分别获取两组小鼠腹股沟处脂肪组织来源的脂肪干细胞,体外纯化扩增至第3代。对细胞进行成骨方向诱导后,使用茜素红染色法检测脂肪干细胞的成骨能力及组间区别；实时荧光定量PCR、免疫荧光染色以及Western Blot检测脂肪干细胞成骨相关基因mRNA和成骨相关蛋白的表达。结果：茜素红染色结果显示,骨质疏松来源的ASCs钙盐沉积明显少于对照组,而real-time PCR和免疫荧光染色结果均指出,与对照组ASCs相比,实验组的ASCs细胞内成骨相关基因mRNAs的转录以及相关蛋白的表达量均显著降低。以上结果表明,与对照组相比,去势后的小鼠来源的脂肪干细胞成骨向分化能力降低(P0.05)。结论：1.通过切除雌性小鼠双侧卵巢,成功构建出骨质疏松症动物模型；2.来自去势组的OP-ASCs较假手术组的ASCs增殖能力显著降低；3.OP-ASCs成骨分化能力较正常ASCs显著降低。
[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.
【学位授予单位】：四川医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580

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