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

转染人SOD基因的细胞对呼吸窘迫综合征进行基因治疗的药物研究

• 批准号: 04454543
• 负责人: OKUMURA Katsuhiko
• 金额: $ 3.58万
• 依托单位: KOBE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454543/
• 关键词: Superoxide dismutase; Gene therapy; SOD cDNA; Culture cell; Superoxide; Secretion; Non-secretion; Catalase; SOD cDNA; 形質転換細胞; スーパーオキサイドジスムターゼ; ラット; cDNA; 呼吸窮迫症候群; PCR

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.

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。将人Cu，Zn-SOD cDNA与人白细胞介素-2（IL-2）基因的前导序列在病毒启动子的控制下融合，构建了表达载体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。超氧化物处理后，这些转染细胞中的过氧化脂质浓度低于正常细胞，这些结果表明，细胞中SOD cDNA的转染对超氧化物诱导的细胞毒性有效。

项目成果

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