發(fā)布時(shí)間：2018-06-29 18:47

  本文選題：膝關(guān)節(jié)骨性關(guān)節(jié)炎 + γ谷氨酰基羧化酶�。� 參考：《南昌大學(xué)》2015年碩士論文

【摘要】：研究目的:探討慢病毒介導(dǎo)的γ谷氨�；然�(Vitamin K dependent gamma-glutamyl carboxylase,GGCX)基因?qū)ν霉切躁P(guān)節(jié)炎軟骨細(xì)胞凋亡的影響,以及對(duì)白細(xì)胞介素-1β(Interleukin-1β,IL-1β)、腫瘤壞死因子(Tumor necrosis factor,TNF)表達(dá)的影響,從而為GGCX基因運(yùn)用于骨性關(guān)節(jié)炎臨床治療提供理論依據(jù)。研究方法:1、選取12只雄性日本大耳兔采用膝關(guān)節(jié)前交叉韌帶切斷術(shù)構(gòu)建模型,6周后將構(gòu)建好模型兔處死,并進(jìn)行軟骨細(xì)胞的分離,培養(yǎng),運(yùn)用Ⅱ型膠原纖維染色和Alcian Blue染色法對(duì)培養(yǎng)的軟骨細(xì)胞進(jìn)行鑒定。2、將培養(yǎng)好的軟骨細(xì)胞隨機(jī)分成三組:空白對(duì)照組(不作處理)、陰性對(duì)照組(轉(zhuǎn)染不含GGCX慢病毒)、轉(zhuǎn)染組(轉(zhuǎn)染含GGCX慢病毒)。運(yùn)用qRT-PCR及Western blot檢測(cè)各組軟骨細(xì)胞中GGCX、Ⅱ型膠原、TNF、IL-1βmRNA及蛋白表達(dá)水平。3、采用Annexin V-FITCPI法檢測(cè)上述各組細(xì)胞凋亡情況。研究結(jié)果:1、第1代骨性關(guān)節(jié)炎兔軟骨細(xì)胞呈多角形扁平狀,可有多個(gè)突起,Ⅱ型膠原纖維染色后可見(jiàn)細(xì)胞及細(xì)胞外基質(zhì)呈黃色或棕黃色,Alcian Blue染色后可見(jiàn)細(xì)胞呈現(xiàn)天藍(lán)色。2、qRT-PCR和Western blot檢測(cè)結(jié)果示:轉(zhuǎn)染組GGCX及Ⅱ型膠原m RNA及蛋白水平表達(dá)較空白對(duì)照組及陰性對(duì)照組明顯升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),空白對(duì)照組及陰性對(duì)照組無(wú)明顯差異(P0.05);轉(zhuǎn)染組TNF、IL-1βmRNA及蛋白水平表達(dá)較空白對(duì)照組及陰性對(duì)照組明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05),空白對(duì)照組及陰性對(duì)照組無(wú)明顯差異(P0.05)。3、Annexin V-FITCPI法檢測(cè)各組凋亡率結(jié)果示:轉(zhuǎn)染組凋亡率為(17.833±2.577)%,較空白對(duì)照組(25.467±2.511)%及陰性對(duì)照組(26.167±2.616)%明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05),空白對(duì)照組及陰性對(duì)照組無(wú)明顯差異(P0.05)。結(jié)論:1、膝關(guān)節(jié)前交叉韌帶切斷術(shù)可以成功構(gòu)建骨性關(guān)節(jié)炎模型,并可以分離及培養(yǎng)出骨性關(guān)節(jié)炎軟骨細(xì)胞;2、轉(zhuǎn)染含GGCX慢病毒可以明顯增加軟骨細(xì)胞GGCX、Ⅱ型膠原的表達(dá);3、轉(zhuǎn)染含GGCX慢病毒可以明顯降低軟骨細(xì)胞TNF、IL-1β的表達(dá);4、轉(zhuǎn)染含GGCX慢病毒后可以抑制骨性關(guān)節(jié)炎軟骨細(xì)胞凋亡。
[Abstract]:Objective: to investigate the effect of Vitamin K dependent gamma-glutamyl carboxylase GGCX gene on the apoptosis of chondrocytes in rabbits with osteoarthritis and the expression of interleukin-1 尾 (IL-1 尾) and tumor necrosis factor (TNF) in rabbit osteoarthritis. Therefore, the application of GGCX gene in the clinical treatment of osteoarthritis provides theoretical basis. Methods: 1. 12 male Japanese large ear rabbits were selected to construct the model by anterior cruciate ligament (ACL) resection of the knee. The rabbits were killed after 6 weeks, and the chondrocytes were separated and cultured. The cultured chondrocytes were identified by type 鈪，

本文編號(hào)：2083017