本文選題：流體剪切力 + 骨保護(hù)素　； 參考：《蘭州大學(xué)》2010年碩士論文

【摘要】： 目的：探索流體剪切力(Fluid Shear Stress, FSS)作用下,兩種調(diào)節(jié)骨骼重建的重要分子骨保護(hù)素(osteoprotegerin, OPG)和破骨細(xì)胞分化因子(osteoclast differentiation factor, ODF)即細(xì)胞核因子kB受體活化因子配體(receptor activator of NF-kB ligand, RANKL)的蛋白表達(dá)情況,從而揭示流體剪切力對(duì)于維持骨組織形成與吸收動(dòng)態(tài)平衡的特殊作用。 方法：本研究采用體外模型對(duì)MC3T3-E1細(xì)胞加載流體剪切力,經(jīng)過不同時(shí)長(zhǎng)(Omin、30min、60min、90min、120min)加力后,對(duì)MC3T3-E1細(xì)胞分別進(jìn)行了染色和裂解,然后運(yùn)用免疫熒光和蛋白印跡法對(duì)骨保護(hù)素(OPG)和破骨細(xì)胞分化因子(RANKL)的蛋白表達(dá)水平進(jìn)行定量分析。 結(jié)果：靜態(tài)培養(yǎng)組RANKL的表達(dá)隨時(shí)間的延長(zhǎng)而增加,OPG的表達(dá)則減少,使得OPG與RANKL的比值(OPG/RANKL)不斷下降,促使骨組織向著加速吸收的方向發(fā)展；加載流體剪切力30min后,蛋白檢測(cè)發(fā)現(xiàn)FSS促進(jìn)了OPG蛋白表達(dá)且抑制了RANKL蛋白表達(dá),阻滯了OPG/RANKL比值減少的趨勢(shì)：加力60min后,發(fā)現(xiàn)FSS進(jìn)一步促進(jìn)了OPG的表達(dá),同時(shí)RANKL的表達(dá)則受到較大抑制,與靜態(tài)組相比此時(shí)OPG/RANKL的比值增加了50%左右,使得骨組織向著成骨的方向發(fā)展；在120min內(nèi),隨著FSS作用時(shí)間的延長(zhǎng)OPG/RANKL的比值也在不斷的增加,各組間OPG與RANKL的比值(OPG/RANKL)有顯著的提高(P0.05)。 結(jié)論：成骨細(xì)胞對(duì)FSS的響應(yīng)要經(jīng)歷一個(gè)短暫的潛伏期,但與其他作用力相比流體剪切力對(duì)成骨細(xì)胞的作用更加有效；流體剪切力作用使得OPG的表達(dá)增加,RANKL的表達(dá)減少,二者的協(xié)同作用(OPG/RANKL)在成骨細(xì)胞和破骨細(xì)胞聯(lián)合調(diào)節(jié)骨骼形成和吸收的過程中扮演著重要角色,從而進(jìn)一步說明FSS對(duì)OPG和RANKL的作用影響著骨組織形成與吸收的動(dòng)態(tài)平衡。
[Abstract]:Objective: to explore the effect of fluid Shear Stress, FSS) on shear stress. The protein expression of osteoprotegerin (OPG) and osteoclast differentiation factor, ODF), the nuclear factor kB receptor activator ligand receptor of NF-kB ligand (RANKLL), are two important molecules regulating bone remodeling. Thus, the special role of fluid shear stress in maintaining the dynamic balance of bone formation and absorption is revealed. Methods: in this study, the MC3T3-E1 cells were stained and lysed with fluid shear force (FSF) loaded with the model in vitro, and the MC3T3-E1 cells were treated with the stress for 30 min, 90 min or 120 min respectively, and the MC3T3-E1 cells were stained and lysed respectively after the stress was applied to the MC3T3-E1 cells for 30 min, 90 min and 120 min, respectively. Then the protein expression levels of osteoprotegerin (OPG) and osteoclast differentiation factor (RANKL) were quantitatively analyzed by immunofluorescence and Western blot. Results: in the static culture group, the expression of RANKL increased and decreased with the increase of time, and the ratio of OPG to RANKL decreased, and the bone tissue developed towards the direction of accelerated absorption. Protein detection showed that FSS promoted the expression of OPG protein and inhibited the expression of RANKL protein, and blocked the decrease of OPG/RANKL ratio. After adding 60min, it was found that FSS further promoted the expression of OPG, while the expression of RANKL was inhibited greatly. Compared with the static group, the ratio of OPG/RANKL increased by about 50%, which made the bone tissue develop toward the direction of osteogenesis, and in 120min, the ratio of OPG to RANKL increased with the prolongation of the time of FSS, and the ratio of OPG to RANKL increased significantly (P 0.05). Conclusion: the response of osteoblasts to FSS has a short incubation period, but fluid shear stress is more effective than other forces on osteoblasts, and fluid shear stress can increase the expression of OPG and decrease the expression of RANKL. The synergistic action of OPG / RANKL plays an important role in the process of osteoblasts and osteoclasts jointly regulating bone formation and absorption, which further indicates that the effect of FSS on OPG and RANKL affects the dynamic balance of bone formation and absorption.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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