PHARMACEUTICAL STUDY FOR GENE THERAPY USING CELL TRANSFECTED WITH HUMAN SOD GENE TO RESPIRATORY DISTRESS SYNOROME

将人Cu，Zn-SOD（superoxidedismutase）cDNA分别插入真核表达载体pRc/RSV和pRc/CMV，用脂质体法转染大鼠皮肤成纤维细胞（FR细胞）、大鼠皮肤原代培养细胞和大鼠肺上皮细胞（L2细胞）。转染细胞中各SOD浓度均升高。这些转染细胞抵抗百草枯合成的超氧化物处理和黄嘌呤/黄嘌呤氧化酶法诱导的细胞杀伤。构建了人Cu，Zn-SOD cDNA与人白细胞介素-2（IL-2）基因前导序列在病毒启动子控制下融合的表达载体（IL-SOD CMV）。转染该载体的FR细胞和L2细胞在培养液中合成并分泌SOD。这些转染细胞抵抗超氧化物诱导的细胞杀伤处理。经超氧化物处理后，转染细胞内脂质过氧化物浓度明显低于正常细胞，表明转染SOD cDNA的细胞对超氧化物诱导的细胞毒性有明显的抑制作用。

Human Cu, Zn-SOD (superoxide dismutase) cDNA was inserted in eukaryotic expression vectors (pRc/RSV and pRc/CMV), and then those vectors were transfected in rat skin fibroblast cell (FR cell), rat skin primary culture cell, and rat lung epitherial cell (L2 cell) with Lipofection method. Each SOD concentration in transfected cells was increased. These transfected cells resisted to a treatment of superoxide synthesized with Paraquart and xanthin/xanthin oxidase methods induced cell killing. After treatment of superoxide, the lipid peroxide concentration in these transfected cells was less than that of normal cells.Expression vector (IL-SOD CMV) was constructed containing human Cu, Zn-SOD cDNA fused in a frame to a leader sequence of human interleukin-2 (IL-2) gene under the control of a viral promoter. FR cell and L2 cell transfected with this vector synthesized and secreted SOD in culture medium. These transfected cells resisted to a treatment of superoxide induced cell killing. The lipid peroxide concentration in these transfected cells was less than normal cells after treatment of superoxide.These results show that the transfection of SOD cDNA in cell was effective in the superoxide induced cytotoxicity.