本文選題：功能性電刺激 + 肌衛(wèi)星細(xì)胞　； 參考：《南方醫(yī)科大學(xué)》2014年碩士論文

【摘要】：研究背景及意義： 功能性電刺激原理產(chǎn)生的各種起搏器正逐漸應(yīng)用于臨床,而肌肉的起搏器如麻痹的環(huán)杓后肌喉起搏器、麻痹的膀胱逼尿肌起搏器和正常神經(jīng)支配的吞咽肌起搏器等已進(jìn)入臨床實(shí)驗(yàn)并在臨床中使用。目前,起搏器帶來(lái)的電刺激對(duì)肌肉包括麻痹肌肉和正常肌肉的影響還不明確。肌肉起搏器在補(bǔ)償肌肉功能的同時(shí)也導(dǎo)致肌肉本身的變化,肌衛(wèi)星干細(xì)胞作為肌纖維細(xì)胞的主要來(lái)源,在電刺激的影響下對(duì)肌肉的再生影響尚不十分明確。近年來(lái),研究發(fā)現(xiàn)肌衛(wèi)星干細(xì)胞是肌肉再生的主要來(lái)源,而電刺激對(duì)肌衛(wèi)星干細(xì)胞的研究也非常少。 頭頸肌是發(fā)聲、呼吸、吞咽及頭頸部運(yùn)動(dòng)的重要肌肉。支配頭頸肌的神經(jīng)發(fā)生不可逆性損傷后,肌肉發(fā)生不可逆性萎縮。衛(wèi)星干細(xì)胞作為肌肉細(xì)胞的主要來(lái)源,是肌肉再生和修復(fù)的關(guān)鍵。因此,我們以電刺激頭頸肌衛(wèi)星干細(xì)胞為研究立足點(diǎn)展開工作。本研究擬通過(guò)電刺激體外培養(yǎng)衛(wèi)星干細(xì)胞,了解其活性、增殖和分化的變化。以期為衛(wèi)星干細(xì)胞增殖提供有利的刺激手段,并為衛(wèi)星干細(xì)胞的增殖和分化機(jī)制研究提供一定的基礎(chǔ),更希望為臨床肌肉起搏器的進(jìn)一步研發(fā)提供實(shí)驗(yàn)室基礎(chǔ)。 目的： 利用小鼠肢體來(lái)源的C2C12細(xì)胞系及比格犬的頦舌肌衛(wèi)星干細(xì)胞體外培養(yǎng)作為研究對(duì)象,使用固定電壓、固定刺激時(shí)間、不同脈寬和不同頻率的脈沖電培養(yǎng)細(xì)胞,了解不同模式電刺激對(duì)衛(wèi)星干細(xì)胞的增殖和分化能力的影響。 方法： 1.取體外培養(yǎng)的肌源性C2C12細(xì)胞株,采用固定電壓(5V)、固定刺激模式、不同脈寬(Oms, lms,2ms,4ms,8ms,20ms六組)的串刺激(5s,3s-on,2s-off)細(xì)胞系,以及固定電壓(5V)、固定刺激模式、不同頻率(Opps、2pps、10pps、20pps、40pps、80pps六組)使用MTT比色法、流式細(xì)胞周期檢測(cè)、qRT-PCR觀察細(xì)胞的活性、增殖及分化。 2.取體外培養(yǎng)的比格犬頦舌肌衛(wèi)星干細(xì)胞體外培養(yǎng),采用固定電壓(5V)、固定脈寬、不同頻率(Opps、2pps、10pps、40pps、80pps、160六組)的串刺激(5s,3s-on,2s-off)刺激細(xì)胞,使用MTT比色法、流式細(xì)胞周期檢測(cè)和Western blot觀察細(xì)胞的活性、增殖及分化。 3.用SPSS19.0統(tǒng)計(jì)軟件進(jìn)行分析。采用Spearman相關(guān)分析了解刺激脈寬及頻率的增加同細(xì)胞活性、增殖和分化狀態(tài)的相關(guān)性。采用Kruskal-Wallis H非參數(shù)檢驗(yàn)方法比較組間差異。以P0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1.隨著脈沖電刺激的脈寬增大會(huì)導(dǎo)致肌源性C2C12細(xì)胞的活性降低(r=-0.724,P0.01),分化能力有提高,在脈寬20ms時(shí)影響較大(p=0.044)；隨著電刺激的頻率加快,肌源性C2C12細(xì)胞的增殖(r=0.643,P=0.004)和分化能力提高(r=0.637,P=0.004；r=0.660,P=0.003),分化的肌細(xì)胞亞型有從快型向慢型轉(zhuǎn)變的趨勢(shì),在80hz時(shí)較明顯(P0.05)。 2.隨著電刺激的頻率加快,頦舌肌衛(wèi)星干細(xì)胞活性和增殖沒有變化,分化能力明顯提高,肌細(xì)胞亞型從快型向慢型轉(zhuǎn)變,在頻率為160hz變化明顯(P0.01)。 結(jié)論：體外實(shí)驗(yàn)中,隨著脈沖電刺激的脈寬增大會(huì)導(dǎo)致衛(wèi)星干細(xì)胞的活性降低,分化能力加強(qiáng),肌細(xì)胞亞型從快型向慢型轉(zhuǎn)變。隨著電刺激的頻率加快,衛(wèi)星干細(xì)胞的分化能力提高,肌細(xì)胞亞型從快型向慢型轉(zhuǎn)變。
[Abstract]:Research background and significance:
Various pacemakers produced by the principle of functional electrical stimulation are gradually applied to the clinic, while the muscle pacemaker such as paralytic posterior dipper pacemaker, paralyzed bladder detrusor pacemaker and normal denervation pacemaker have entered clinical trials and used in clinical practice. At present, the electrical stimulation of pacemaker brings to muscle package. It is not clear that the effects of paralytic muscles and normal muscles are not clear. Muscle pacemakers also cause muscle changes while compensating muscle function. Muscle satellite stem cells are the main source of muscle fiber cells. The effect of electrical stimulation on muscle regeneration is not very clear. In recent years, the study found that muscle satellite stem cells are muscles. The main source of regeneration is the electrical stimulation of muscle satellite stem cells.
The head and neck muscles are important muscles of the vocal, respiratory, swallowing, and head and neck movement. After the irreversible damage to the neurogenesis of the head and neck muscles, the muscle is irreversible. As the main source of the muscle cells, the satellite stem cells are the key to the muscle regeneration and repair. Therefore, we use electric stimulation of the head and neck muscle satellite stem cells as the research foothold. The purpose of this study is to stimulate the activity, proliferation and differentiation of satellite stem cells by electrical stimulation in vitro, in order to provide a favorable stimulus for the proliferation and differentiation of satellite stem cells, and to provide a basis for the study of the mechanism of proliferation and differentiation of satellite stem cells, and hope to provide a further research and development of the clinical muscle pacemaker. Laboratory basis.
Objective:
The effects of different modes of electrical stimulation on the proliferation and differentiation of satellite stem cells were investigated by using the C2C12 cell lines from the limbs of the mice and the culture of the genial muscle satellite stem cells of the Beagle in vitro.
Method錛，

本文編號(hào)：1954251