机械拉伸对大鼠成骨细胞生理活性及IGF-1、力生长因子表达的影响
发布时间：2018-10-18 06:39
【摘要】：本文采用生物力学、细胞生物学、分子生物学等方法,研究机械拉伸刺激对成骨细胞生理活性和生长因子表达的影响,重点就力学信号——基因表达关系进行了初步探索。主要工作和结论如下: ①采用消化法从初生大鼠的颅盖骨中分离、纯化、培养和鉴定成骨细胞。 ②使用周期性拉伸装置和静态拉伸装置对大鼠颅盖骨成骨细胞施加机械拉伸应变,应变水平均为15%,受载时间为6、12、24、36h,研究同一应变水平的机械拉伸刺激对成骨细胞在不同加载时间生化响应以及基因表达的影响。 1) 应用流式细胞技术研究机械拉伸应变对成骨细胞增殖的影响,探讨此类细胞在力学生理背景下的生长行为。结果表明:同一应变水平(15%)的周期性拉伸和静态拉伸下,随着受载时间的增加,成骨细胞的相对增殖指数增加,在各加载时间点周期性拉伸下成骨细胞相对增殖指数都大于静态拉伸下成骨细胞的相对增殖指数,但变化趋势相似。在受载6h 后,成骨细胞的相对增殖指数最大,而在受载12h、24h、36h 后,成骨细胞相对增殖指数下降,特别是在受载36h 后,细胞的相对增殖指数趋向于1。说明机械拉伸能促进细胞增殖,并且这种促进作用与受载时间相关。而且周期性机械拉伸的促进作用比静态机械拉伸的作用更强,就相对增殖指数而言,受载的初期(6h),对成骨细胞增殖的影响最明显。 2) 同一应变水平(15%)周期性拉伸与静态拉伸作用下,受载成骨细胞碱性磷酸酶相对活力和胞外无机钙质的相对分泌量显示,静态拉伸下成骨细胞在受载初期(6h),与对照组相比,相对碱性磷酸酶活力和胞外无机钙质的相对分泌量均略小于1,随受载时间的增加逐渐升高且均大于1。相对碱性磷酸酶活力36h 达最高;相对胞外钙分泌量24h 达最高,36h 后略有下降,但仍大于1。而周期性拉伸下的成骨细胞,各受载时间点相对碱性磷酸酶活力与相对胞外钙分泌量都小于1。说明短期静态拉伸刺激能抑制成骨细胞的分化、矿化,较长时间的静态拉伸刺激能促进成骨细胞的分化、矿化。而周期性拉伸刺激对成骨细胞的分化、矿化起抑制作用。 3) 通过半定量RT-PCR 方法分析了机械拉伸对成骨细胞IGF-1、MGF mRNA表达的影响。结果显示:在周期性拉伸组、静态拉伸组和对照组中IGF-1 都有一定水平的表达,相比较而言,周期性拉伸组细胞表达最强。MGF 在周期性拉伸组细胞中的表达明显高于静态拉伸组细胞的表达,对照组几乎没有表达。而在各受载时间组中IGF-1 mRNA 、MGF mRNA 的表达量都是周期性拉伸组细胞明显强于静态拉伸组细胞。MGF mRNA 在6h 后达到最大值,IGF-1 mRNA 在12h 后达到最
[Abstract]:In this paper, biomechanics, cell biology and molecular biology were used to study the effects of mechanical tensile stimulation on the physiological activity and the expression of growth factors in osteoblasts, with emphasis on the relationship between mechanical signal-gene expression. The main work and conclusions are as follows: 1 digested and purified from the skull of newborn rats. Culture and identification of osteoblasts. (2) Mechanical tensile strain was applied to osteoblasts of rat calvaria with cyclic and static stretching devices. The effects of mechanical tensile stimulation at the same strain level on the biochemical response and gene expression of osteoblasts at different loading times were studied. 1) the effects of mechanical tensile strain on proliferation of osteoblasts were studied by flow cytometry. To study the growth behavior of this kind of cells under the background of mechanics and physiology. The results showed that the relative proliferation index of osteoblasts increased with the increase of loading time at the same strain level (15%). The relative proliferation index of osteoblasts under cyclic stretching was higher than that under static tension, but the trend was similar. The relative proliferation index of osteoblasts was the highest after loading for 6 h, while that of osteoblasts decreased after being loaded for 12 h or 24 h or 36 h, especially after being loaded for 36 h, the relative proliferation index of osteoblasts tended to be 1. The results showed that mechanical stretching could promote cell proliferation, and the effect was related to loading time. And the effect of periodic mechanical stretching is stronger than that of static mechanical stretching, in terms of relative proliferation index, At the early stage of loading (6h), the proliferation of osteoblasts was affected most obviously. 2) under the same strain level (15%), the cells were subjected to cyclic and static stretching. The relative activity of alkaline phosphatase (ALP) and the relative secretion of extracellular calcium in osteoblasts were compared with those in the control group. The activity of relative alkaline phosphatase and the relative secretion of extracellular inorganic calcium were slightly less than 1, and gradually increased with the increase of loading time. The relative alkaline phosphatase activity reached the highest level at 36h, and the relative extracellular calcium secretion reached the highest level at 24h, and decreased slightly after 36h, but still more than 1. The relative alkaline phosphatase activity and extracellular calcium secretion of osteoblasts under cyclic stretching were less than 1. The results showed that the short-term static tensile stimulation could inhibit the differentiation of osteoblasts and mineralization, and the long time static tension stimulation could promote the differentiation and mineralization of osteoblasts. Periodic tensile stimulation inhibited the differentiation and mineralization of osteoblasts. 3) the effect of mechanical stretching on IGF-1,MGF mRNA expression in osteoblasts was analyzed by semi-quantitative RT-PCR. The results showed that there was a certain level of IGF-1 expression in the periodic stretching group, the static stretching group and the control group. The expression of MGF was significantly higher in the periodic stretching group than in the static stretching group, but almost no expression in the control group. However, the expression of IGF-1 mRNA, MGF mRNA was significantly stronger in the cyclically stretched group than in the static stretch group at 6 h, and the maximum expression of IGF-1 mRNA was reached at 12 h after each loading time.
【学位授予单位】：重庆大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329
本文编号：2278306
[Abstract]:In this paper, biomechanics, cell biology and molecular biology were used to study the effects of mechanical tensile stimulation on the physiological activity and the expression of growth factors in osteoblasts, with emphasis on the relationship between mechanical signal-gene expression. The main work and conclusions are as follows: 1 digested and purified from the skull of newborn rats. Culture and identification of osteoblasts. (2) Mechanical tensile strain was applied to osteoblasts of rat calvaria with cyclic and static stretching devices. The effects of mechanical tensile stimulation at the same strain level on the biochemical response and gene expression of osteoblasts at different loading times were studied. 1) the effects of mechanical tensile strain on proliferation of osteoblasts were studied by flow cytometry. To study the growth behavior of this kind of cells under the background of mechanics and physiology. The results showed that the relative proliferation index of osteoblasts increased with the increase of loading time at the same strain level (15%). The relative proliferation index of osteoblasts under cyclic stretching was higher than that under static tension, but the trend was similar. The relative proliferation index of osteoblasts was the highest after loading for 6 h, while that of osteoblasts decreased after being loaded for 12 h or 24 h or 36 h, especially after being loaded for 36 h, the relative proliferation index of osteoblasts tended to be 1. The results showed that mechanical stretching could promote cell proliferation, and the effect was related to loading time. And the effect of periodic mechanical stretching is stronger than that of static mechanical stretching, in terms of relative proliferation index, At the early stage of loading (6h), the proliferation of osteoblasts was affected most obviously. 2) under the same strain level (15%), the cells were subjected to cyclic and static stretching. The relative activity of alkaline phosphatase (ALP) and the relative secretion of extracellular calcium in osteoblasts were compared with those in the control group. The activity of relative alkaline phosphatase and the relative secretion of extracellular inorganic calcium were slightly less than 1, and gradually increased with the increase of loading time. The relative alkaline phosphatase activity reached the highest level at 36h, and the relative extracellular calcium secretion reached the highest level at 24h, and decreased slightly after 36h, but still more than 1. The relative alkaline phosphatase activity and extracellular calcium secretion of osteoblasts under cyclic stretching were less than 1. The results showed that the short-term static tensile stimulation could inhibit the differentiation of osteoblasts and mineralization, and the long time static tension stimulation could promote the differentiation and mineralization of osteoblasts. Periodic tensile stimulation inhibited the differentiation and mineralization of osteoblasts. 3) the effect of mechanical stretching on IGF-1,MGF mRNA expression in osteoblasts was analyzed by semi-quantitative RT-PCR. The results showed that there was a certain level of IGF-1 expression in the periodic stretching group, the static stretching group and the control group. The expression of MGF was significantly higher in the periodic stretching group than in the static stretching group, but almost no expression in the control group. However, the expression of IGF-1 mRNA, MGF mRNA was significantly stronger in the cyclically stretched group than in the static stretch group at 6 h, and the maximum expression of IGF-1 mRNA was reached at 12 h after each loading time.
【学位授予单位】：重庆大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329
【引证文献】
相关硕士学位论文 前1条
1 王峰;不同负荷运动对老年大鼠IGF-I表达及骨骼生物力学性能的影响[D];河北师范大学;2011年
，本文编号：2278306
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