發(fā)布時(shí)間：2018-03-23 18:02

  本文選題：流體剪切力　切入點(diǎn)：MC3T3-E1成骨細(xì)胞　出處：《蘭州大學(xué)》2014年碩士論文

【摘要】：目的：隨著中國(guó)逐步進(jìn)入老齡化社會(huì),老年人的健康問(wèn)題成為全社會(huì)的一大焦點(diǎn),骨質(zhì)疏松由于其發(fā)病率高而成為一個(gè)公共衛(wèi)生問(wèn)題,目前主要是通過(guò)藥物以及運(yùn)動(dòng)完成骨質(zhì)疏松的治療,但由于目前對(duì)于骨骼感受應(yīng)力的機(jī)制不是很明確,導(dǎo)致臨床醫(yī)生無(wú)法正確地指導(dǎo)患者進(jìn)行合理的運(yùn)動(dòng)促進(jìn)康復(fù),因此本研究應(yīng)用自制的體外流體剪切力(Fluid shear stress, FSS)加載裝置用生理強(qiáng)度的流體剪切力加載成骨細(xì)胞(MC3T3-E1),明確ERK5信號(hào)通路的傳導(dǎo)機(jī)制,以及對(duì)成骨有很重要作用的兩個(gè)因子骨保護(hù)素(osteoprotegerin, OPG)和破骨細(xì)胞分化因子(Osteoclast differentiation factor, ODF),又稱細(xì)胞核因子κB受體活化因子配體(receptor activator of NF-κB ligand, RANKL)的表達(dá)情況。以期能夠?yàn)榕R床骨質(zhì)減少性疾病患者正確合理的運(yùn)動(dòng)提供理論依據(jù)。 方法：將ERK5特異性阻斷劑BIX02188配制成不同濃度,并干預(yù)MC3T3-E1成骨細(xì)胞,后用MTT法檢測(cè)490nm OD值,觀察成骨細(xì)胞的增殖狀態(tài),并運(yùn)用熒光定量PCR檢測(cè)ERK5mRNA的表達(dá)情況。用生理強(qiáng)度為12dyne/cm2的流體剪切力加載成骨細(xì)胞,后檢測(cè)OPG、RANKL mRNA的表達(dá)情況。先用之前實(shí)驗(yàn)得出的濃度BIX02188干預(yù)成骨細(xì)胞,之后再給成骨細(xì)胞加載生理強(qiáng)度為12dyne/cm的流體剪切力,檢測(cè)OPG、RANKL mRNA的表達(dá)情況。 結(jié)果：流體剪切力促進(jìn)成骨細(xì)胞的增殖；濃度為15μM的ERK5特異阻斷劑BIX02188能夠有效的抑制ERK5mRNA的表達(dá)；MTT結(jié)果證實(shí)：ERK5特異性阻斷劑阻斷ERK5活化后,成骨細(xì)胞增殖受到明顯抑制；強(qiáng)度為12dyne/cm的流體剪切力能夠促進(jìn)成骨細(xì)胞OPG mRNA表達(dá)(P0.05),降低RANKL mRNA表達(dá)(P0.05)；當(dāng)BIX02188干預(yù)成骨細(xì)胞后,流體剪切力對(duì)成骨細(xì)胞OPG、RANKL mRNA表達(dá)的影響明顯減弱(P0.05)。 結(jié)論：生理強(qiáng)度為12dyne/cm2的流體剪切力能促進(jìn)成骨細(xì)胞增殖,對(duì)OPG mRNA的表達(dá)有促進(jìn)作用,對(duì)RANKL mRNA的表達(dá)有抑制作用；ERK5能夠有效介導(dǎo)流體剪切力對(duì)成骨細(xì)胞OPG. RANKL mRNA的表達(dá)。
[Abstract]:Objective: as China has gradually entered an aging society, the health of the elderly has become a major focus in the whole society, and osteoporosis has become a public health problem due to its high incidence. At present, the treatment of osteoporosis is mainly done through medicine and exercise. However, because the mechanism of bone stress is not very clear at present, the clinicians are unable to correctly guide the patients to carry out reasonable exercise to promote rehabilitation. Therefore, in order to clarify the transduction mechanism of ERK5 signaling pathway, a self-made in vitro fluid shear stress (FSS) loading device was used to load osteoblasts MC3T3-E1 with physiological strength of fluid shear force. And the expression of osteoprotegerin (OPG) and osteoclast differentiation factor (Osteoclast differentiation factor), also known as nuclear factor 魏 B receptor activating factor receptor activator of NF- 魏 B ligand (RANKL). The correct and reasonable exercise in patients with osteopenia disease provides theoretical basis. Methods: BIX02188, a specific inhibitor of ERK5, was prepared into different concentrations, and MC3T3-E1 osteoblasts were interfered with. The OD value of 490nm was detected by MTT method, and the proliferation of osteoblasts was observed. Fluorescence quantitative PCR was used to detect the expression of ERK5mRNA. Osteoblasts were loaded with fluid shear force with physiological strength of 12dyne/cm2, and then the expression of RANKL mRNA was detected. Firstly, the concentration of BIX02188 was used to interfere with osteoblasts. Then the osteoblasts were subjected to fluid shear stress with physiological strength of 12dyne/cm to detect the expression of OPGG RANKL mRNA. Results: the proliferation of osteoblasts was promoted by fluid shear stress, and the proliferation of osteoblasts was inhibited by BIX02188, a specific inhibitor of ERK5 at concentration of 15 渭 M, which could effectively inhibit the expression of ERK5mRNA. The results showed that the proliferation of osteoblasts was significantly inhibited after the activation of ERK5 was blocked by the specific blocker of 1: ERK5. The fluid shear stress with 12dyne/cm strength could promote the expression of OPG mRNA and decrease the expression of RANKL mRNA in osteoblasts, but the effect of fluid shear stress on the expression of mRNA in osteoblasts was significantly weakened after BIX02188 intervention. Conclusion: fluid shear stress with physiological strength of 12dyne/cm2 can promote the proliferation of osteoblasts, promote the expression of OPG mRNA, and inhibit the expression of RANKL mRNA. ERK5 can effectively mediate the expression of osteoblast OPG. RANKL mRNA by fluid shear stress.
【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】：2014
【分類號(hào)】：R318.01

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