本文選題：成骨樣細(xì)胞MG63 + 壓應(yīng)力�。� 參考：《吉林大學(xué)》2014年碩士論文

【摘要】：目的與背景: 牙槽骨是骨組織中可塑性最強(qiáng)、代謝最活躍者。生理情況下，有牙牙合牙槽骨保持破骨與成骨的動(dòng)態(tài)平衡，使骨組織處于相對(duì)穩(wěn)定的狀態(tài)。長(zhǎng)期缺牙的患者，牙槽骨缺乏牙齒的功能性刺激，而發(fā)生進(jìn)行性、累積性、不可逆轉(zhuǎn)的骨吸收。牙槽骨的不斷吸收是引起臨床很多修復(fù)體失敗的主要原因。應(yīng)力因素是影響剩余牙槽嵴吸收的主要因素，也是我們?nèi)藶榈哪軌虿扇「深A(yù)措施加以改變的。王景云等學(xué)者發(fā)現(xiàn)，應(yīng)力與牙槽骨的吸收密切相關(guān)；咀嚼壓力過(guò)大會(huì)加速剩余牙槽嵴的吸收；適宜的咀嚼壓力能夠延緩剩余牙槽嵴的吸收。 Sato等學(xué)者從人胚胎cDNA文庫(kù)中發(fā)現(xiàn)了一條新基因，即：膜型基質(zhì)金屬蛋白酶-1（MT1-MMP），其表達(dá)于細(xì)胞膜表面，能夠編碼含582個(gè)氨基酸的蛋白質(zhì)。近年來(lái)研究發(fā)現(xiàn)，作為MMPs家族中被發(fā)現(xiàn)的第一個(gè)膜型MMPs，MT1-MMP在正常骨骼發(fā)育及骨改建過(guò)程中具有重要的作用。MT1-MMP表達(dá)于成骨細(xì)胞和破骨細(xì)胞，不僅其自身與正常骨骼組織發(fā)育及骨改建有著密切的聯(lián)系，尚可通過(guò)激活其他MMPs及細(xì)胞因子而影響骨改建的過(guò)程；MT1-MMP能夠調(diào)控細(xì)胞的機(jī)械敏感性；在成骨細(xì)胞分化為骨細(xì)胞的過(guò)程中，MT1-MMP作為一種骨細(xì)胞生成初期的必須物質(zhì)具有決定性作用。研究MT1-MMP在骨組織中的作用形式及作用途徑，對(duì)了解及掌握骨組織相關(guān)的疾患及研究防治方法具有重要的指導(dǎo)意義。 本實(shí)驗(yàn)運(yùn)用四點(diǎn)彎曲力學(xué)加載儀對(duì)體外培養(yǎng)的MG-63細(xì)胞施加不同強(qiáng)度及不同作用時(shí)間的壓應(yīng)力，利用Western Blot及逆轉(zhuǎn)錄PCR檢測(cè)MT1-MMP的表達(dá)水平，為研究MT1-MMP在成骨細(xì)胞中的作用機(jī)制提供了基礎(chǔ)實(shí)驗(yàn)依據(jù)，進(jìn)一步證實(shí)應(yīng)力作用下的細(xì)胞將發(fā)生的細(xì)胞生物學(xué)的改變以及探索咀嚼壓力與剩余牙槽嵴吸收的關(guān)系奠定理論基礎(chǔ)。 實(shí)驗(yàn)方法： 采用四點(diǎn)彎曲力學(xué)加載儀對(duì)培養(yǎng)在細(xì)胞加力板上的MG63細(xì)胞施加壓應(yīng)力，頻率為0.5Hz，設(shè)置梯度變化及單一水平的壓應(yīng)力加載方式，研究壓應(yīng)力在不同力值（0、1000μstrain、2000μstrain、4000μstrain、6000μstrain）及不同時(shí)間（0、1h、3h、6h、9h、12h）對(duì)MT1-MMP蛋白及基因表達(dá)的影響，每組實(shí)驗(yàn)重復(fù)3次，空白對(duì)照組不加力；除應(yīng)力因素外，其他影響因素基本控制一致。 結(jié)果： 1、當(dāng)應(yīng)變值恒定為2000μstrain時(shí)，力學(xué)刺激呈時(shí)間依賴性的上調(diào)成骨樣細(xì)胞MG63細(xì)胞中MT1-MMP蛋白及基因的表達(dá)； 2、當(dāng)應(yīng)變值為2000μstrain時(shí)，力學(xué)刺激對(duì)成骨樣細(xì)胞MG63中MT1-MMP蛋白及基因表達(dá)作用最強(qiáng)，當(dāng)應(yīng)變值為6000μstrain時(shí)，MT1-MMP表達(dá)最低。 結(jié)論： 1、在生理范圍內(nèi)，，當(dāng)對(duì)MG63細(xì)胞施加恒定的壓應(yīng)力時(shí)，MT1-MMP的表達(dá)呈時(shí)間依賴性的上調(diào)； 2、當(dāng)應(yīng)力加載時(shí)間相同時(shí)，MT1-MMP的表達(dá)隨著力值的增大而增加，但應(yīng)力超出生理范圍時(shí)，其表達(dá)則降低。 3、適宜的應(yīng)力在調(diào)節(jié)骨改建過(guò)程中有著重要的作用。
[Abstract]:Purpose and background: Alveolar bone is the most plastic and metabolized bone tissue. Under physiological conditions, the alveolar bone of edentulous teeth keeps the dynamic balance between osteoclast and osteogenesis, which makes the bone tissue relatively stable. In patients with chronic dental deficiency, alveolar bone lacks functional stimulation and develops progressive, cumulative, irreversible bone resorption. The incessant absorption of alveolar bone is the main cause of the failure of many clinical prostheses. Stress factor is the main factor affecting the absorption of residual alveolar ridge. Wang Jingyun and other scholars found that stress is closely related to alveolar bone absorption; masticatory pressure is excessive to accelerate the absorption of residual alveolar ridge; appropriate masticatory pressure can delay the absorption of residual alveolar ridge. Sato and other researchers have found a new gene from the human embryo cDNA library, namely, the membrane matrix metalloproteinase-1 (MMP-1) MT1-MMPN, which is expressed on the surface of the cell membrane and encodes a protein containing 582 amino acids. In recent years, it has been found that MT1-MMP, the first membrane type of MMPsMMP found in MMPs family, plays an important role in normal bone development and bone remodeling. MT1-MMP is expressed in osteoblasts and osteoclasts. Not only is it closely related to normal bone tissue development and bone remodeling, but it can also affect the process of bone remodeling by activating other MMPs and cytokines. MT1-MMP can regulate the mechanical sensitivity of cells. MT1-MMP plays a decisive role in the process of osteoblast differentiation into osteocytes. It is of great significance to understand and master the diseases related to bone tissue and to study the methods of prevention and treatment by studying the forms and ways of action of MT1-MMP in bone tissue. In this study, four point bending mechanical loading apparatus was used to apply compressive stress of different intensity and time to MG-63 cells in vitro. The expression of MT1-MMP was detected by Western Blot and reverse transcription PCR. It provides a basic experimental basis for studying the mechanism of MT1-MMP acting on osteoblasts, further confirms the changes of cell biology under stress and explores the relationship between masticatory pressure and residual alveolar ridge absorption. Experimental methods: A four-point bending mechanical loading instrument was used to apply compressive stress to the MG63 cells cultured on the cell afterloading plate at a frequency of 0.5 Hz, and a gradient change and a single level of compressive stress loading mode were set. The effects of compressive stress on the expression of MT1-MMP protein and gene were studied at different stress values (0 1 000 渭 strain (2000 渭 strain) and 4 000 渭 strain) and at different times. The results showed that the MT1-MMP protein and gene expression were repeated 3 times in each group, but not in the blank control group. Except stress factors, the other influencing factors were basically controlled. Results: 1. When the strain value was 2000 渭 strain, mechanical stimulation up-regulated the expression of MT1-MMP protein and gene in MG63 cells in a time-dependent manner. 2. When the strain value was 2000 渭 strain, the expression of MT1-MMP protein and gene in osteoblast like cell MG63 was the strongest by mechanical stimulation, and the expression of MT1-MMP was the lowest when the strain value was 6000 渭 strain. Conclusion: 1. In physiological range, the expression of MT1-MMP was up-regulated in a time-dependent manner when MG63 cells were subjected to constant compressive stress. 2. When the stress loading time is the same, the expression of MT1-MMP increases with the increase of stress, but when the stress exceeds the physiological range, the expression of MT1-MMP decreases. 3. Suitable stress plays an important role in regulating bone remodeling.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R783

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