本文關(guān)鍵詞： 間充質(zhì)干細(xì)胞 成骨分化 脈沖電磁場(chǎng) 凋亡 骨髓間充質(zhì)干細(xì)胞 拉曼光譜 脈沖電磁場(chǎng) 干細(xì)胞 分子生物學(xué) 光鑷　出處：《廣西醫(yī)科大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：背景采用低頻脈沖電磁場(chǎng)干預(yù)骨髓間充質(zhì)干細(xì)胞增殖分化的研究很多,但高頻(380MHz)脈沖電磁場(chǎng)干預(yù)的研究國(guó)內(nèi)少有報(bào)道。 目的觀察380 MHz高頻脈沖電磁場(chǎng)照射對(duì)骨髓間充質(zhì)干細(xì)胞增殖分化的影響。 方法分離培養(yǎng)SD大鼠骨髓間充質(zhì)干細(xì)胞,取第3代細(xì)胞隨機(jī)分為4組：A為化學(xué)試劑成骨誘導(dǎo)組、B為化學(xué)試劑成骨誘導(dǎo)并電磁照射組、C為電磁場(chǎng)照射組、D為空白對(duì)照組。觀察各組骨髓間充質(zhì)干細(xì)胞培養(yǎng)過(guò)程中細(xì)胞形態(tài)、數(shù)量、總蛋白量等方面變化。 結(jié)果與未經(jīng)高頻脈沖電磁場(chǎng)照射組相比,經(jīng)高頻脈沖電磁場(chǎng)照射后,骨髓間充質(zhì)干細(xì)胞未見明顯分化。電磁場(chǎng)照射組細(xì)胞增殖速度、總蛋白含量均低于空白對(duì)照組(P0.05)。電磁照射組細(xì)胞凋亡率較空白對(duì)照組增加(P0.05)。 結(jié)論高頻脈沖電磁場(chǎng)對(duì)骨髓間充質(zhì)干細(xì)胞的成骨分化趨勢(shì)不明顯,可抑制其增殖,促進(jìn)其凋亡。 背景中低頻脈沖電磁場(chǎng)干預(yù)骨髓間充質(zhì)干細(xì)胞增殖分化的研究很多,但高頻電磁場(chǎng)照射干細(xì)胞的研究罕有報(bào)道,拉曼光譜方法分析電磁場(chǎng)照射干細(xì)胞的研究更為稀少。 目的利用拉曼光譜觀察并分析380 MHz高頻脈沖電磁場(chǎng)照射骨髓間充質(zhì)干細(xì)胞后的細(xì)胞生物學(xué)影響。 方法本實(shí)驗(yàn)體外分離培養(yǎng)SD大鼠骨髓間充質(zhì)干細(xì)胞,將第3代細(xì)胞隨機(jī)接種到六孔板,分2組：無(wú)電磁組D、380MHz脈沖電磁照射組C,采集7天后細(xì)胞的拉曼光譜。 結(jié)果①C、D組干細(xì)胞拉曼光譜特征峰基本相同,經(jīng)origin軟件作圖后波形基本相同；②380MHz組BMSCs拉曼光譜峰強(qiáng)度普遍下降且有多處特征峰幾乎消失,如524,650,764,1302,1367,1561cm-1等；③C、D組拉曼光譜都出現(xiàn)了骨骼組織的必須成分Hydroxyapatite特征峰(966 cm-1),hydoxyproline特征峰(1206 cm-1),但C、D無(wú)明顯差異。 結(jié)論①光鑷?yán)庾V技術(shù)能夠應(yīng)用于單個(gè)干細(xì)胞分子層面的生物化學(xué)變化的檢測(cè)和研究；②用拉曼光譜技術(shù)監(jiān)測(cè)電磁場(chǎng)對(duì)細(xì)胞的影響是一種可行的方法。
[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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R329

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