本文選題：股骨頭骨壞死 + 動(dòng)脈灌注��； 參考：《中國人民解放軍醫(yī)學(xué)院》2017年博士論文

【摘要】：一、目的:擬在研究人股骨頭骨壞死標(biāo)本骨結(jié)構(gòu)的基礎(chǔ)上,探索壞死股骨頭內(nèi)miRNA-214表達(dá)與局部成、破骨細(xì)胞活性之間的關(guān)系及潛在機(jī)制,制備腺相關(guān)病毒搭載miRNA-214抑制劑,觀察藥物調(diào)節(jié)成骨、破骨活性防治大鼠股骨頭骨壞死塌陷的效果。二、方法:①2015年3月至2016年5月期間,收集我院16例股骨頭骨壞死標(biāo)本。標(biāo)本進(jìn)行Micro-CT掃描后,根據(jù)骨質(zhì)密度的不同,將每個(gè)標(biāo)本分為不同區(qū)域,分別采用病理學(xué)檢測、實(shí)時(shí)熒光定量PCR、WesternBlot等方法對骨微觀結(jié)構(gòu)及成骨、破骨細(xì)胞活性進(jìn)行分析。PCR檢測股骨頭骨壞死標(biāo)本骨組織,觀察不同區(qū)域成骨、破骨相關(guān)基因表達(dá)情況及miRNA-214表達(dá)情況。②制備以腺相關(guān)病毒為載體的miRNA-214抑制劑藥物,評價(jià)其體外調(diào)節(jié)成骨、破骨的能力。③建立大鼠股骨頭骨壞死模型,觀察AAV-anti-miRNA-214藥物的治療效果。三、結(jié)果:①標(biāo)本壞死區(qū)域miRNA-214表達(dá)顯著增高,破骨活性顯著增高,成骨活性顯著降低。而硬化區(qū)成骨活性顯著增高,破骨活性顯著降低。同時(shí),miRNA-214在股骨頭骨壞死標(biāo)本壞死區(qū)域顯著增高;②成功制備AAV-anti-miRNA-214,發(fā)現(xiàn)其能夠明顯抑制破骨細(xì)胞的功能,促進(jìn)成骨細(xì)胞的功能,同時(shí)可以顯著下調(diào)miRNA-214的表達(dá),TRAP染色結(jié)果顯示AAV-anti-miRNA-214可以顯著抑制骨髓間充質(zhì)干細(xì)胞向破骨細(xì)胞分化的能力。MC3T3-E1 細(xì)胞加入AAV病毒包裹的anti-miRNA-214 , 發(fā)現(xiàn)AAV-anti-miRNA-214能夠明顯促進(jìn)成骨細(xì)胞的功能,ALP染色的結(jié)果顯示成骨細(xì)胞的功能增強(qiáng);③AAV-anti-miRNA-214可以促進(jìn)成骨活性,抑制破骨活性,防治大鼠股骨頭骨壞死塌陷。四、結(jié)論:AAV- anti-miR-214可以促進(jìn)成骨活性,抑制破骨活性,其雙向調(diào)節(jié)作用可有效防治大鼠早期股骨頭骨壞死塌陷。
[Abstract]:Objective: to explore the relationship between the expression of miRNA-214 and local osteoclast and osteoclast activity in the necrotic femoral head on the basis of studying the bone structure of the osteonecrosis specimen of human femoral head, and to prepare the adeno-associated virus carrying miRNA-214 inhibitor. To observe the effect of drug regulating osteogenesis and osteoclast activity preventing and treating osteonecrosis of femoral head in rats. Methods: from March 2015 to May 2016, 16 cases of osteonecrosis of femoral head were collected. After Micro-CT scanning, each specimen was divided into different regions according to the different bone density. The microstructure and osteogenesis of bone were detected by pathological examination and real-time fluorescence quantitative PCR Western blot. Osteoclast activity was analyzed. PCR was used to detect bone tissue of osteonecrosis specimens of femoral head. Osteogenesis, osteoclast-associated gene expression and miRNA-214 expression were observed in different regions. 2 miRNA-214 inhibitor drugs were prepared using adeno-associated virus as vector. To evaluate the ability of osteoregulation and osteoclast in vitro to establish osteonecrosis model of femoral head in rats and to observe the therapeutic effect of AAV-anti-miRNA-214 drugs. Results: the expression of miRNA-214, osteoclast activity and osteogenesis activity in the necrotic region were significantly increased and osteogenic activity was significantly decreased. However, osteogenic activity in sclerotic area was significantly increased and osteoclast activity was significantly decreased. At the same time, AAV-anti-miRNA-214 was successfully prepared by miRNA-214 in the necrotic area of osteonecrosis of femoral head. It was found that AAV-anti-miRNA-214 could significantly inhibit the function of osteoclasts and promote the function of osteoblasts. At the same time, the expression of miRNA-214 was significantly down-regulated by trap staining. The results showed that AAV-anti-miRNA-214 could significantly inhibit the ability of bone marrow mesenchymal stem cells to differentiate into osteoclasts. MC3T3-E1 cells were added to anti-miRNA-214 wrapped with AAV virus. It was found that AAV-anti-miRNA-214 could significantly promote osteoblasts. The results of ALP staining showed that the function of osteoblasts was enhanced. 3AAV-anti-miRNA-214 can promote osteogenic activity, inhibit osteoclast activity and prevent osteonecrosis of femoral head in rats. Conclusion: AAV- anti-miR-214 can promote osteogenic activity and inhibit osteoclast activity, and its bidirectional regulation can effectively prevent and cure early osteonecrosis of femoral head in rats.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R681.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 薛紅芳;吳維光;趙容;;下調(diào)microRNA-214表達(dá)對人卵巢癌SKOV-3細(xì)胞遷移及侵襲的影響[J];中國腫瘤生物治療雜志;2016年04期

2 王仁俊;溫美玲;未曉巍;武慧;李華;郝錫聯(lián);齊云峰;周曉馥;;MicroRNA-214在慢性心衰大鼠心肌細(xì)胞鈣調(diào)控中的作用與機(jī)制研究[J];吉林師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2016年01期

3 楊玉寶;劉洪亞;闞金慶;高忠禮;李林;;股骨頭壞死患者壞死骨組織中miRNA表達(dá)譜變化[J];山東醫(yī)藥;2015年34期

4 周桃玉;陳崇;溫旺榮;;microRNA-214在女性乳腺癌患者血清中的表達(dá)及臨床意義[J];臨床檢驗(yàn)雜志;2015年04期

5 王東;魏文平;王芳;;MicroRNA-214在缺氧所致心肌細(xì)胞損傷過程中的作用[J];心臟雜志;2014年04期

6 張麗靜;吳晨鵬;張志勇;樊智彬;賀新奇;劉博;趙增仁;;MiR-214對人結(jié)腸癌HCT116細(xì)胞生物學(xué)功能的影響[J];腫瘤;2013年11期

7 張麗靜;吳晨鵬;張志勇;劉博;樊智彬;裴永彬;趙增仁;;MicroRNA-214在結(jié)直腸癌中的表達(dá)及其對細(xì)胞增殖的影響[J];世界華人消化雜志;2013年27期

8 趙增仁;樊智彬;張麗靜;吳晨鵬;張學(xué)明;劉博;;結(jié)直腸癌中microRNA-214的表達(dá)及其與p53的關(guān)系[J];廣東醫(yī)學(xué);2013年08期

，

本文編號：1850110