退变腰椎间盘细胞经体外转基因培养生物学活性的实验研究

目的构建转化生长因子（transforming growth factor，TGF-β3的真核表达载体pEGFP-TGF-β3转染椎间盘髓核细胞，研究转基因TGF-β3对退变髓核细胞生物学特性的影响。方法通过手术方法制作椎间盘突变模型，从而获取原代退变椎间盘髓核细胞，通过脂质体将真核载体pEGFP-TGF-β3导入髓核细胞，然后对细胞的形态和增殖活性（MTT法）进行观察，应用Westen Blot检测TGF-β3在髓核细胞的表达含量，应用免疫细胞化学方法检测转染后髓核细胞的Ⅱ型胶原的表达。结果髓核细胞转染后，细胞活性增强，TGF-β3表达增加，并随着时间的延长而增加。Ⅱ型胶原表达增加。结论TGF-β3转染退变髓核细胞可起到维持髓核细胞表型，并在细胞传代后仍发挥调节作用。TGF-β3确实具有促进髓核细胞增殖和Ⅱ型胶原合成的能力，从而有可能延缓甚至逆转椎间盘退变。

Study on biologic activity of degenerated intervertebral disc cells by transgenic culture in vitro

Objective To study the proliferating effects of liposome-mediated transforming growth factor-β3 （ TGF- β3 ） gene on degenerated rabbit nucleus pulposus cells. Methods Rabbit intervertebral disc degeneration models were built by surgical then to gain the degenerated nucleus pulposus cells, pEGFP-TGF-β3 were transduced into the nucleus pulposus cells via liposome. The figure and proliferating activity of the cells were observed, and the quantity of TGF-β3 in the ceils were determined by Western Blot and the expression of collagen Ⅱwas evaluated by immunoehemieal staining. Results Both the proliferating activity and the quantity of TGF-β3 enhanced after gene were transduced and the expression of collagen Ⅱ increased. Conclusion TGF-β3 gene mediated by liposome could persistently express in degenerated nucleus pulposus cells, and produce biological effects on nucleus both in cell proliferating and synthesis of collagen Ⅱ, and furthermore prevent or reverse the process of disc degeneration.