AAV_2-hVEGF_(165)与AAV_2-hTGFβ_1对退变兔椎间盘纤维环细胞的生物学效应  

The biological effect of AAV_2-hVEGF_(165) and AAV_2-hTGFβ_1 on degenerative annulus fibrosus cells of rabbit intervertebral disc

作　　者：董跃福[1] 牟志芳[2] 石法亮[1] 胡有谷[3] 

机构地区：[1]胶南市人民医院骨二科,山东青岛266400 [2]胶南市人民医院ICU,青岛266400 [3]青岛大学医学院附属医院骨科,266000

出　　处：《颈腰痛杂志》2008年第3期209-212,共4页The Journal of Cervicodynia and Lumbodynia

基　　金：国家自然科学基金资助(项目号:30271318)

摘　　要：目的采用基因治疗方法,把AAV2-hVEGF165和AAV2-hTGFβ1共转染兔纤维环细胞,观察其生物学活性,并进一步评价人血管内皮生长因子16(5hVEGF165)和人转化生长因子β(1hTGFβ1)逆转椎间盘退变的可行性。方法分离并培养兔椎间盘纤维环细胞,以腺相关病毒(AAV2)为载体,分别携带hVEGF165和hTGFβ1cDNA,转染兔纤维环细胞。用Western blotting检测hVEGF165和hTGFβ1的表达,并用同样的方法检测纤维环细胞中Ⅰ型胶原的表达。结果AAV2能够感染兔椎间盘细胞。用Western blotting可以检测到AAV2-hVEGF165和AAV2-hTGFβ1在退变椎间盘细胞中的表达,并且,两种因子联合转染纤维环细胞时Ⅰ型胶原的表达要比分别转染是显著增高。结论hVEGF165和hTGFβ1共同转染培养的兔退变纤维环细胞,Ⅰ型胶原的表达显著增高。Objective To observe the bioactivity of cotransduce the AAV2-hVEGF165 and AAV2- hTGFβ1 to rabbit annulus fibrosus cells. Further to evaluate the possibility of hVEGF165 and hTGFβ1 to reverse intervertebral disc degeneration by gene therapy. Methods The annulus fibrosus cells from rabbit intervertebral discs were isolated and cultured and then transduced with adeno-associated virus2（AAV2）vector encoding both hVEGF165 cDNA and hTGFβ1 cDNA or respectively. Then the expression of hVEGF165 and hTGFβ1 were detected by Western blotting,and the expression of collagen I in annulus fibrosus cells was also explored by the same method. Results AAV2 can infect the rabbit annulus fibrous cells. The expression of AAV2-hVEGF165 and AAV2-TGFβ1 were manifested in degenerative annulus fibrosus cells detected by Western blotting,and the expression of collagen I in annulus fibrosus cells cotransfected by AAV2-hVEGF165 and AAV2-hTGFβ1 was markedly increased than that of monogenic transfected cells. Conclusions With the delivery of both hVEGF165 and hTGF-β1 genes simultaneously,the collagen Ⅰ synthesis were increased in cultured degenerated annulus fibrosus cells.

关 键 词：血管内皮生长因子 转化生长因子β1 腺相关病毒 Ⅰ型胶原 

分 类 号：Q527.2[生物学—生物化学] Q954.633