本文選題：流體剪切力 + 骨保護素��； 參考：《蘭州大學》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素(osteoprotegerin, OPG)和破骨細胞分化因子(osteoclast differentiation factor, ODF)即細胞核因子kB受體活化因子配體(receptor activator of NF-kB ligand, RANKL)的蛋白表達情況,從而揭示流體剪切力對于維持骨組織形成與吸收動態(tài)平衡的特殊作用。 方法：本研究采用體外模型對MC3T3-E1細胞加載流體剪切力,經(jīng)過不同時長(Omin、30min、60min、90min、120min)加力后,對MC3T3-E1細胞分別進行了染色和裂解,然后運用免疫熒光和蛋白印跡法對骨保護素(OPG)和破骨細胞分化因子(RANKL)的蛋白表達水平進行定量分析。 結(jié)果：靜態(tài)培養(yǎng)組RANKL的表達隨時間的延長而增加,OPG的表達則減少,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,蛋白檢測發(fā)現(xiàn)FSS促進了OPG蛋白表達且抑制了RANKL蛋白表達,阻滯了OPG/RANKL比值減少的趨勢：加力60min后,發(fā)現(xiàn)FSS進一步促進了OPG的表達,同時RANKL的表達則受到較大抑制,與靜態(tài)組相比此時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作用時間的延長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：成骨細胞對FSS的響應要經(jīng)歷一個短暫的潛伏期,但與其他作用力相比流體剪切力對成骨細胞的作用更加有效；流體剪切力作用使得OPG的表達增加,RANKL的表達減少,二者的協(xié)同作用(OPG/RANKL)在成骨細胞和破骨細胞聯(lián)合調(diào)節(jié)骨骼形成和吸收的過程中扮演著重要角色,從而進一步說明FSS對OPG和RANKL的作用影響著骨組織形成與吸收的動態(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.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R329

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