高频脉冲电磁场对骨髓间充质干细胞增殖分化的影响

发布时间：2018-02-26 22:42

本文关键词： 间充质干细胞成骨分化脉冲电磁场凋亡骨髓间充质干细胞拉曼光谱脉冲电磁场干细胞分子生物学光镊　出处：《广西医科大学》2011年硕士论文　论文类型：学位论文

【摘要】：背景采用低频脉冲电磁场干预骨髓间充质干细胞增殖分化的研究很多,但高频(380MHz)脉冲电磁场干预的研究国内少有报道。目的观察380 MHz高频脉冲电磁场照射对骨髓间充质干细胞增殖分化的影响。方法分离培养SD大鼠骨髓间充质干细胞,取第3代细胞随机分为4组：A为化学试剂成骨诱导组、B为化学试剂成骨诱导并电磁照射组、C为电磁场照射组、D为空白对照组。观察各组骨髓间充质干细胞培养过程中细胞形态、数量、总蛋白量等方面变化。结果与未经高频脉冲电磁场照射组相比,经高频脉冲电磁场照射后,骨髓间充质干细胞未见明显分化。电磁场照射组细胞增殖速度、总蛋白含量均低于空白对照组(P0.05)。电磁照射组细胞凋亡率较空白对照组增加(P0.05)。结论高频脉冲电磁场对骨髓间充质干细胞的成骨分化趋势不明显,可抑制其增殖,促进其凋亡。背景中低频脉冲电磁场干预骨髓间充质干细胞增殖分化的研究很多,但高频电磁场照射干细胞的研究罕有报道,拉曼光谱方法分析电磁场照射干细胞的研究更为稀少。目的利用拉曼光谱观察并分析380 MHz高频脉冲电磁场照射骨髓间充质干细胞后的细胞生物学影响。方法本实验体外分离培养SD大鼠骨髓间充质干细胞,将第3代细胞随机接种到六孔板,分2组：无电磁组D、380MHz脉冲电磁照射组C,采集7天后细胞的拉曼光谱。结果①C、D组干细胞拉曼光谱特征峰基本相同,经origin软件作图后波形基本相同；②380MHz组BMSCs拉曼光谱峰强度普遍下降且有多处特征峰几乎消失,如524,650,764,1302,1367,1561cm-1等；③C、D组拉曼光谱都出现了骨骼组织的必须成分Hydroxyapatite特征峰(966 cm-1),hydoxyproline特征峰(1206 cm-1),但C、D无明显差异。结论①光镊拉曼光谱技术能够应用于单个干细胞分子层面的生物化学变化的检测和研究；②用拉曼光谱技术监测电磁场对细胞的影响是一种可行的方法。
[Abstract]:Background there are many studies on the effects of low-frequency pulsed electromagnetic fields on the proliferation and differentiation of bone marrow mesenchymal stem cells, but there are few reports on the effects of high-frequency pulsed electromagnetic fields on the proliferation and differentiation of bone marrow mesenchymal stem cells. Objective to observe the effect of 380 MHz high frequency pulsed electromagnetic field irradiation on the proliferation and differentiation of bone marrow mesenchymal stem cells. Methods SD rat bone marrow mesenchymal stem cells were isolated and cultured. The third generation of cells were randomly divided into 4 groups: group B was induced by chemical reagent, group B was induced by chemical reagent, group C was irradiated by electromagnetic field, and group D was irradiated by electromagnetic field. The morphology of bone marrow mesenchymal stem cells was observed during the culture of bone marrow mesenchymal stem cells in each group. Changes in quantity, total protein content, etc. Results Bone marrow mesenchymal stem cells did not differentiate significantly after high frequency pulsed electromagnetic field irradiation compared with those without high frequency pulsed electromagnetic field irradiation. The proliferation rate of bone marrow mesenchymal stem cells in the electromagnetic field group was higher than that in the electromagnetic field group. The total protein content was lower than that of the blank control group (P 0.05), and the apoptosis rate of the electromagnetic irradiation group was higher than that of the blank control group (P 0.05). Conclusion the high frequency pulsed electromagnetic field has no obvious tendency of osteogenic differentiation of bone marrow mesenchymal stem cells, which can inhibit the proliferation and promote the apoptosis of bone marrow mesenchymal stem cells. In the background, there have been many studies on the effects of low-frequency pulsed electromagnetic fields on the proliferation and differentiation of bone marrow mesenchymal stem cells, but there are few reports on the high-frequency electromagnetic field irradiation of stem cells. Aim to observe and analyze the effects of 380 MHz high frequency pulsed electromagnetic field on cell biology of bone marrow mesenchymal stem cells (BMSCs) by Raman spectroscopy. Methods Sprague-Dawley rat bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured in vitro. The third passage cells were randomly inoculated into the six-hole plate. The cells were divided into two groups: no electromagnetic group (DX 380MHz) pulsed electromagnetic irradiation group (C), and the Raman spectra of the cells were collected after 7 days. Results 1the Raman characteristic peaks of stem cells in group C ~ (2 +) D were basically the same, and the intensity of Raman spectra of BMSCs in group C ~ (2 380) MHz was almost the same after drawing by origin software, and a number of characteristic peaks almost disappeared. For example, in the Raman spectra of 524, 650, 7602, 13676, 1561cm-1, and so on, the essential Hydroxyapatite characteristic peak of bone tissue, 966 cm-1, hydoxyproline, 1206 cm-1, was found, but there was no significant difference in CUD. Conclusion 1 Raman spectroscopy of optical tweezers can be used to detect and study the biochemical changes of single stem cell at molecular level. It is a feasible method to monitor the effect of electromagnetic field on cells by Raman spectroscopy.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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