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目的研究人胰岛素基因表达质粒转染鼠成纤维细胞（Ｌｔｋ－）后对糖尿病大鼠血糖的影响。方法重组的人胰岛素基因表达质粒通过脂质体法转染鼠成纤维细胞（Ｌｔｋ－），经Ｇ４１８选择性筛选获得一株较高胰岛素表达水平的细胞株（Ｌｔｋ－／ＰＲＩ１２），大量扩增，直接注射至糖尿病大鼠腹腔内，并与转染空载体的对照Ｌｔｋ－细胞比较，观察在糖尿病大鼠体内的胰岛素表达及对血糖等变化的影响。结果空载体转染的对照细胞与未转染组相似，无胰岛素表达，植入糖尿病大鼠腹腔后，血糖仍持续稳定在较高水平，体重进行性下降；而接受胰岛素表达载体细胞的糖尿病大鼠，在观察的２１天内血Ｃ肽持续表达，血糖明显下降，体重逐渐恢复。结论人胰岛素基因能成功转染非胰岛β细胞并表达目的基因使血糖水平下降。本研究为进一步开展糖尿病基因治疗研究奠定了基础。
Objective To investigate the effect of human insulin gene expression plasmid on the blood glucose of diabetic rats after transfection with murine fibroblasts (Ltk-). Methods Recombinant human insulin gene expression plasmid was transfected into murine fibroblasts (Ltk-) by lipofectamine. A cell line with high expression level of insulin (Ltk- / PRI-12) was screened by G418, Were injected into the peritoneal cavity of diabetic rats and compared with control Ltk- cells transfected with empty vector to observe the effect of insulin on blood glucose and other changes in diabetic rats. Results Compared with the untransfected group, the control cells transfected with empty vector showed no insulin expression. After transplanted into the peritoneal cavity of diabetic rats, the blood glucose level remained stable at a high level and the weight decreased progressively. However, the number of diabetic rats receiving insulin expression vector was large Rats in the observed 21 days of continuous expression of blood C-peptide, decreased blood glucose, body weight gradually restored. Conclusion The human insulin gene can successfully transfect non-islet β cells and express the target gene to decrease the blood glucose level. This study laid the foundation for further research on gene therapy of diabetes.