本文關(guān)鍵詞：Wnt信號通路在壓力刺激下大鼠髁突軟骨病理變化過程中的作用及其機(jī)制研究　出處：《南京大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 大鼠髁突軟骨 骨關(guān)節(jié)炎 軟骨細(xì)胞 細(xì)胞增殖 Wnt信號通路 骨關(guān)節(jié)炎樣變 細(xì)胞外基質(zhì) 炎癥反應(yīng) Wnt

【摘要】：第一部分Wnt信號通路在壓力刺激下大鼠髁突軟骨細(xì)胞增殖過程中的作用[目的]研究軟骨細(xì)胞增殖活力受到抑制的原因及其在髁突軟骨病理性變薄過程中的作用,本研究擬探討Wnt信號通路對壓力刺激下的大鼠髁突軟骨病理性變薄,軟骨細(xì)胞增殖水平受到抑制的影響,明確其具體的作用和機(jī)制。[方法]選擇6周齡SD雄性大鼠應(yīng)用本課題組自主設(shè)計的顳下頜關(guān)節(jié)壓應(yīng)力加載動物模型裝置(專利號：201120210396.4),對大鼠髁突軟骨施加向上、向后的80g/側(cè)超負(fù)荷壓應(yīng)力刺激誘導(dǎo)大鼠髁突軟骨發(fā)生病理性變薄,將所有動物設(shè)為早期組和晚期組,每組分為四個亞組：空白對照組,單純加藥組,單純加力組和加力加藥組。蘇木素-伊紅(HE)染色觀察大鼠髁突軟骨層厚度及形態(tài)學(xué)的變化;免疫組化(immunohistochemistry, IHC)檢測Wnt信號通路下游標(biāo)記分子β-catenin和GSK3β,以及細(xì)胞增殖標(biāo)記分子cyclin D1, PCNA, Histone H3的表達(dá)改變。采用RT-PCR檢測Wnt下游標(biāo)志分子Axin2在mRNA水平的表達(dá)改變。[結(jié)果]1.髁突軟骨受到機(jī)械力刺激的晚期,Wnt信號通路受損。2.機(jī)械力刺激髁突軟骨的晚期,應(yīng)用GSK3β抑制劑可以一定程度恢復(fù)Wnt信號通路的活力,并顯著提高髁突軟骨細(xì)胞的增殖活力。3.機(jī)械力刺激髁突軟骨的晚期,GSK3β抑制劑通過上調(diào)cyclin D1, PCNA和Histone H3的表達(dá),促進(jìn)細(xì)胞的DNA合成,恢復(fù)軟骨細(xì)胞的增殖水平。[結(jié)論]1.機(jī)械力刺激髁突軟骨導(dǎo)致Wnt信號通路受損,軟骨細(xì)胞的增殖活力受到抑制及髁突軟骨病理性變薄。2.應(yīng)用GSK3β抑制劑恢復(fù)Wnt信號通路的活力可以上調(diào)軟骨細(xì)胞的增殖水平并緩解髁突軟骨的病理性變薄。第二部分Wnt信號通路在壓力刺激下大鼠髁突軟骨骨關(guān)節(jié)炎樣病理變化過程中的作用[目的]為了研究Wnt信號通路對大鼠髁突軟骨在持續(xù)機(jī)械應(yīng)力作用下病理性改變的作用,我們對已經(jīng)證實(shí)調(diào)節(jié)軟骨細(xì)胞增殖活力的Wnt信號通路進(jìn)一步研究其在維持細(xì)胞外基質(zhì)內(nèi)環(huán)境穩(wěn)態(tài)和炎癥反應(yīng)中的作用。探討Wnt信號通路在大鼠髁突軟骨骨關(guān)節(jié)炎樣病理變化過程中的作用及其機(jī)制。[方法]選擇6周齡SD雄性大鼠建立髁突軟骨骨關(guān)節(jié)炎動物模型,同樣分為早期組和晚期組,每組分為四個亞組：空白對照組,單純加藥組,單純加力組合加力加藥組。采用阿辛藍(lán)(Ab)染色觀察大鼠髁突軟骨蛋白多糖含量的變化；免疫組化(immunohistochemistry, IHC)檢測Col-Ⅱ和NF-α的蛋白水平變化,RT-PCR檢測Col-Ⅱ和Col-X型膠原蛋白,蛋白酶MMPs,炎癥因子TNF-α和IL-1的mRNA水平的表達(dá)改變,研究膠原蛋白和多糖表達(dá)水平,以及炎癥因子表達(dá)水平的變化。[結(jié)果]1.GSK3β抑制劑恢復(fù)機(jī)械應(yīng)力加載大鼠髁突軟骨晚期的Wnt信號通路活力,抑制MMPs的活性并上調(diào)膠原蛋白合蛋白多糖的表達(dá)。2.GSK3β抑制劑還顯著下調(diào)了機(jī)械力刺激晚期的軟骨細(xì)胞中炎癥因子TNF-α和IL-1的高表達(dá)。[結(jié)論]機(jī)械力刺激髁突軟骨抑制了Wnt信號通路,導(dǎo)致軟骨細(xì)胞外基質(zhì)受損和炎癥應(yīng)答的激活。GSK3β抑制劑恢復(fù)受損的Wnt信號通路活力,并維持軟骨細(xì)胞外基質(zhì)內(nèi)環(huán)境的穩(wěn)態(tài),抑制一系列炎癥反應(yīng)。
[Abstract]:The first part of the Wnt signaling pathway in the process of pressure stimulated proliferation of rat condylar cartilage cells [Objective] to study the proliferation of cartilage cells inhibited and the reasons in the condylar cartilage pathological thinning process, this paper intends to discuss the Wnt pathway thorn Condylar Cartilage Pathological rats induced by the pressure becomes thin, the proliferation level of cartilage cells inhibited the effect, clear its function and mechanism. The specific methods of selection of 6 week old male SD rats using the research group designed the temporomandibular joint stress loading animal model device (Patent No.: 201120210396.4), the rat condylar cartilage is applied to on the back side, 80g/ overload stress induced rat condylar Cartilage Pathological thinning, all animal for early and late groups, each group was divided into four subgroups: blank control group, simple dosing group, simple Afterburner afterburner group and drug group. Hematoxylin and eosin (HE) staining was used to observe the changes of condylar cartilage thickness and morphology of rats; immunohistochemistry (immunohistochemistry, IHC) detection of Wnt signaling pathways downstream molecular markers of beta -catenin and GSK3 beta cell proliferation marker cyclin, D1, PCNA, Histone H3 expression the detection of Wnt by RT-PCR. Expression of Axin2 in mRNA downstream molecular level changes. Results the]1. condylar cartilage by mechanical stimulation of the late Wnt pathway impaired.2. mechanical stimulation of the condylar cartilage in the late application of GSK3 beta inhibitors can be recovered to a certain degree of Wnt signaling pathway activity, and significantly improve the condylar cartilage the cell proliferation activity of.3. mechanical stimulation of the condylar cartilage in the late GSK3 beta inhibitors through upregulation of cyclin expression of D1, PCNA and Histone H3, promote the synthesis of DNA cells, restore the proliferation of chondrocytes. [conclusion]1. mechanical stimulation of condylar cartilage impairs Wnt signaling, cartilage cell proliferation activity was inhibited and the condylar cartilage pathological thinning of.2. application of GSK3 beta Wnt signaling pathway inhibitor to restore vitality can change the pathological increase of cartilage cell proliferation level and ease of condylar cartilage is thin. The second part of the Wnt signaling pathway in the process of pressure under the stimulation of condylar cartilage in osteoarthritis pathological changes of rats [Objective] to study the signal transduction of Wnt in rat condylar cartilage under continuous mechanical stress change of force under the action of Wnt signaling pathway we have proven to regulate cartilage cell proliferation activity further its role in maintaining cell homeostasis and inflammation in stroma. To investigate the Wnt signal pathway in the process of condylar cartilage of osteoarthritis like pathological changes in rats. Method: effect and mechanism To establish the condylar cartilage of osteoarthritis animal model of 6 week old male SD rats were equally divided into early and late groups, each group was divided into four subgroups: blank control group, simple dosing group, single drug group. The combination of afterburner afterburner alcian blue (Ab) staining to observe the changes of rat condylar cartilage bone proteoglycan content; immunohistochemistry (immunohistochemistry, IHC) changes in protein levels of Col- II and NF- alpha detection, RT-PCR detection and Col- II type Col-X collagen, protease MMPs, change the expression of inflammatory factor TNF- alpha and IL-1 mRNA levels of collagen, protein and polysaccharide expression levels and inflammatory factors the expression level of]1.GSK3. The beta Wnt signaling pathway inhibitor restored the mechanical dynamic loading of condylar cartilage in rats with the expression of.2.GSK3 beta inhibitors inhibit MMPs activity and increase of collagen and proteoglycan also significantly reduced Mechanical stimulation of cartilage cells in the late inflammatory factor TNF- alpha and IL-1 expression. Conclusion] mechanical stimulation of condylar cartilage inhibits the Wnt signaling pathway, Wnt signaling pathway leading to the recovery of activity impaired activation of.GSK3 beta inhibitors of extracellular matrix of cartilage damage and inflammatory response, and maintain the chondrocyte extracellular matrix homeostasis of internal environment a series of inflammatory reaction, inhibition.

【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R782
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本文編號：1386355