本文選題：遠(yuǎn)中缺失同源盒基因5 + 肌節(jié)同源盒基因1��； 參考：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1.顱面骨和髂骨胚胎來源不同,它們之間存在不同的基因表達(dá)模式和骨代謝水平。遠(yuǎn)中缺失同源盒基因(Distal-less homeobox,Dlx)和肌節(jié)同源盒基因(Msh homeobox,Msx)在骨代謝方面發(fā)揮重大作用,尤其是Dlx-5和Msx-1。本研究對(duì)象為長(zhǎng)期接受唑來膦酸藥物的大鼠,通過檢測(cè)其Dlx-5和Msx-1蛋白及基因表達(dá)水平的變化,從而探究唑來膦酸對(duì)顱面骨和髂骨的影響是否存在差異。2.建立雙膦酸鹽相關(guān)性頜骨壞死(Bisphosphonate-related osteonecrosis of the jaw,BRONJ)動(dòng)物模型,觀察BRONJ的臨床表現(xiàn)、影像學(xué)改變及組織病理學(xué)特點(diǎn),探究Dlx-5和Msx-1在BRONJ致病機(jī)制中發(fā)揮的作用。方法:1.36只雌性白化封閉群大鼠(Sprague-Dawley,SD)隨機(jī)分成三組,每組12只。唑來膦酸組腹腔注射唑來膦酸(0.2 mg/kg),每周3次持續(xù)12周。生理鹽水組腹腔注射等量生理鹽水持續(xù)12周,另外一組為對(duì)照組,不做任何處理。利用免疫組織化學(xué)染色、蛋白質(zhì)印跡法(Western blot)和實(shí)時(shí)定量-聚合酶鏈反應(yīng)(Real time-polymerase chain reaction,RT-PCR)檢測(cè)顱面骨(包括上頜骨、下頜骨和頂骨)和髂骨中Dlx-5和Msx-1的表達(dá)水平。2.24只雌性SD大鼠隨機(jī)分為兩組,每組12只。唑來膦酸組腹腔注射唑來膦酸(0.2 mg/kg),每周3次持續(xù)12周。對(duì)照組腹腔注射等量生理鹽水,每周3次持續(xù)12周。12周后所有大鼠麻醉下拔除左下第一磨牙,臨床觀察拔牙創(chuàng)愈合情況。拔牙后8周所有動(dòng)物被處死。對(duì)大鼠左下頜骨進(jìn)行X線片檢查和Micro-CT檢查,探究其影像學(xué)改變。對(duì)下頜骨軟硬組織進(jìn)行蘇木精-伊紅(Hematoxylin-eosin,HE)染色和Masson染色,探究其組織病理學(xué)特點(diǎn),進(jìn)一步確定BRONJ動(dòng)物模型。通過Western blot、RT-PCR技術(shù)檢測(cè)Dlx-5和Msx-1在大鼠下頜骨拔牙創(chuàng)周圍硬組織中的表達(dá)水平。結(jié)果:1.分別與生理鹽水組和對(duì)照組比較,唑來膦酸組上、下頜骨中Dlx-5的蛋白和基因表達(dá)水平升高(P0.05),上、下頜骨中Msx-1的蛋白和基因水平降低(P0.01)。在蛋白和基因水平上,唑來膦酸組髂骨中的Dlx-5和Msx-1的表達(dá)水平均降低(P0.05)。唑來膦酸組與其他兩組比較,頂骨中Dlx-5和Msx-1的蛋白和基因表達(dá)水平均無顯著變化(P0.05),它們之間的差異無統(tǒng)計(jì)學(xué)意義。2.拔牙后8周,臨床觀察發(fā)現(xiàn)唑來膦酸組大鼠拔牙創(chuàng)不愈合,并且實(shí)驗(yàn)動(dòng)物口內(nèi)可見死骨暴露。影像學(xué)檢查及組織病理學(xué)檢查均證實(shí)BRONJ的發(fā)生。Masson染色結(jié)果顯示唑來膦酸組拔牙創(chuàng)周圍牙齦膠原纖維纖細(xì)。Western blot、RT-PCR檢測(cè)結(jié)果顯示唑來膦酸組拔牙創(chuàng)周圍頜骨中Dlx-5蛋白和基因表達(dá)水平均升高(P0.01),同時(shí)Msx-1表達(dá)水平降低(P0.01)。結(jié)論:1.唑來膦酸作用下顱面骨和髂骨中Dlx-5和Msx-1表達(dá)水平的變化趨勢(shì)存在差異性,這在一定程度上表明了唑來膦酸對(duì)大鼠骨骼的影響存在位置差異性。與頂骨和髂骨相比,頜骨對(duì)唑來膦酸的應(yīng)用最為敏感。2.腹腔注射唑來膦酸聯(lián)合拔牙的方法能夠成功誘導(dǎo)大鼠發(fā)生BRONJ樣病變。在BRONJ動(dòng)物模型中Dlx-5和Msx-1表達(dá)水平的變化趨勢(shì)能夠促進(jìn)骨形成,進(jìn)一步破壞骨代謝平衡,導(dǎo)致拔牙創(chuàng)局部骨硬化,其在BRONJ的發(fā)生、發(fā)展過程中發(fā)揮重要作用。
[Abstract]:Objective: 1. craniofacial bone and iliac bone have different embryonic sources, and there are different gene expression patterns and bone metabolism levels between them. Distal-less homeobox (Dlx) and Msh homeobox (Msx) genes (Msh homeobox, Msx) play a major role in bone metabolism, especially in Dlx-5 and Msx-1. based subjects for long-term acceptance of azole By detecting changes in the levels of Dlx-5 and Msx-1 protein and gene expression in the rat, the difference in the effects of zoledronic acid on the craniofacial bone and iliac bone was detected by.2., and the establishment of the bisphosphonate associated osteonecrosis (Bisphosphonate-related osteonecrosis of the jaw, BRONJ) animal model and the clinical manifestations of BRONJ were observed. The role of Dlx-5 and Msx-1 in the pathogenesis of BRONJ was investigated by imaging changes and histopathological features. Methods: 1.36 female albino closed group rats (Sprague-Dawley, SD) were randomly divided into three groups, 12 in each group. Zoledronic acid group was intraperitoneally injected with zoledronic acid (0.2 mg/kg) and lasted 3 times a week for 12 weeks. The normal saline lasted for 12 weeks, and the other group was a control group without any treatment. The expression level of Dlx-5 and Msx-1 in the cranial bone (including the maxilla, mandible and parietal bone) and the iliac bone was detected by immunohistochemical staining, Western blot and real-time quantitative polymerase chain reaction (RT-PCR),.2. .24 female SD rats were randomly divided into two groups, each group was 12. Zoledronic acid group was intraperitoneally injected with zoledronic acid (0.2 mg/kg) and lasted 3 times a week for 12 weeks. The control group was injected with equal amount of saline, 3 times a week after 12 weeks of.12 weeks, all rats were extracted to remove the left inferior molars and observe the healing of tooth extraction at 8 weeks after extraction. The X-ray examination and Micro-CT examination of the left mandible of the rat were carried out to explore the imaging changes. The histopathological features of the soft and hard tissue of the mandible were studied by Hematoxylin-eosin and HE staining and Masson staining, and the model of BRONJ animal was further determined. Dlx-5 and Msx-1 were detected by Western blot and RT-PCR technology. Results: 1. compared with the normal saline group and the control group, 1. the levels of protein and gene expression in the mandible increased (P0.05) in the zoledronic acid group and in the zoledronic acid group (P0.05), and the protein and gene level of Msx-1 in the mandible decreased (P0.01). The expression level of Dlx-5 and Msx-1 in the bone decreased (P0.05). There was no significant change in the protein and gene expression levels of Dlx-5 and Msx-1 in the parietal bone compared with the other two groups (P0.05). There was no significant difference between them, and there was no significant difference between them in.2. after the extraction of teeth 8 weeks after the extraction of the azolidonic acid group. The imaging examination and histopathological examination showed that the.Masson staining results of BRONJ showed that the gingival collagen fibrils were fine.Western blot in the azolidonic acid group, and the RT-PCR detection results showed that the level of Dlx-5 protein and gene expression in the azolidonic acid group was elevated (P0.01) in the surrounding jaw of the azolidonic acid group. The expression level of Msx-1 was reduced (P0.01). Conclusion: there is a difference in the expression of Dlx-5 and Msx-1 in the cranial bone and iliac bone under the action of 1. zoledronic acid, which, to a certain extent, indicates that zoledronic acid has a position difference in the effect of zoledronic acid on the skeleton of rats. Compared with the parietal bone and iliac bone, the maxillofacial application is the most sensitive.2. to the zoledronic acid. Intraperitoneal injection of zoledronic acid combined with tooth extraction can successfully induce BRONJ like lesions in rats. In the BRONJ animal model, the changes in the expression of Dlx-5 and Msx-1 can promote bone formation, further destroy the balance of bone metabolism, and lead to local osseosclerosis, which plays an important role in the occurrence and development of BRONJ.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R782

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 張健;張文怡;任媛媛;;唑來膦酸對(duì)體外破骨細(xì)胞性骨吸收影響的研究[J];口腔頜面修復(fù)學(xué)雜志;2009年06期

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本文編號(hào)：1831304