本文選題：轉化生長因子β3　切入點：基質金屬蛋白酶抑制劑1　出處：《青島大學》2015年碩士論文

【摘要】：目的:探討慢病毒載體介導的轉化生長因子β3(TGFβ3)、基質金屬蛋白酶抑制劑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,從而最終實現延緩甚至逆轉椎間盤退變的目的。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。應用構建的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)學變化,通過Real-time q PCR技術檢測感染后TGFβ3、TIMP1、Ⅱ型膠原和蛋白多糖的m RNA表達量,通過Western blot技術檢測感染后Ⅱ型膠原和蛋白多糖含量,最終評價應用攜帶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代謝的影響。結果:感染復數(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)計學差異。結論:慢病毒載體介導的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)揮生物學效應,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