本文選題：轉(zhuǎn)化生長因子β3　切入點：基質(zhì)金屬蛋白酶抑制劑1　出處：《青島大學(xué)》2015年碩士論文

【摘要】：目的:探討慢病毒載體介導(dǎo)的轉(zhuǎn)化生長因子β3(TGFβ3)、基質(zhì)金屬蛋白酶抑制劑1(TIMP1)聯(lián)合感染對體外培養(yǎng)人退變椎間盤髓核細胞的影響,從而最終實現(xiàn)延緩甚至逆轉(zhuǎn)椎間盤退變的目的。方法:單層培養(yǎng)人退變椎間盤髓核細胞,采用綠色熒光蛋白標記慢病毒(GV287-EGFP)評價其對髓核細胞的感染效率。應(yīng)用構(gòu)建的TGFβ3-P2A-TIMP1慢病毒載體感染髓核細胞,在倒置纖維鏡下觀察細胞形態(tài)學(xué)變化,通過Real-time q PCR技術(shù)檢測感染后TGFβ3、TIMP1、Ⅱ型膠原和蛋白多糖的m RNA表達量,通過Western blot技術(shù)檢測感染后Ⅱ型膠原和蛋白多糖含量,最終評價應(yīng)用攜帶TGFβ3-P2A-TIMP1的慢病毒載體體外感染人退變椎間盤髓核細胞后對其細胞外基質(zhì)代謝的影響。結(jié)果:感染復(fù)數(shù)(MOI)為60時可獲較滿意感染效率。Real-time q PCR示目的基因感染組的TGFβ3、TIMP1、Ⅱ型膠原、蛋白多糖m RNA表達量均明顯高于空病毒感染組及空白對照組(F=16.350~74.378,q=2.010~10.620,P0.05),而空病毒感染組和空白對照組間無統(tǒng)計學(xué)差異。Western blot示目的基因感染組的Ⅱ型膠原、蛋白多糖蛋白表達量均明顯高于空病毒感染組和空白對照組(F=50.495~66.110,q=8.061~16.624,P0.05),而空病毒感染組和空白對照組間無統(tǒng)計學(xué)差異。結(jié)論:慢病毒載體介導(dǎo)的TGFβ3和TIMP1雙基因聯(lián)合感染人椎間盤髓核細胞可長時間穩(wěn)定表達,發(fā)揮生物學(xué)效應(yīng),促進蛋白多糖和Ⅱ型膠原的合成,并最終改善髓核細胞外基質(zhì)的代謝。
[Abstract]:Aim: to investigate the effect of lentivirus vector mediated transforming growth factor 尾 3(TGF 尾 3 and matrix metalloproteinase inhibitor 1 (TIMP 1) co-infection on cultured human degenerative disc nucleus pulposus cells in vitro, so as to delay or even reverse the degeneration of intervertebral disc.Methods: monolayer cultured human degenerative disc nucleus pulposus cells were labeled with green fluorescent protein (GV287-EGFP) to evaluate the infection efficiency of GV287-EGFP on nucleus pulposus cells.The TGF 尾 3-P2A-TIMP1 lentivirus vector was used to infect the nucleus pulposus cells. The morphological changes of the cells were observed under inverted fibroscopy. The expression of m RNA in TGF 尾 3 TIMP 1, type 鈪，

本文編號：1720682