摘要
AIM:To study the expression of human insulin gene in gastrointestinal tracts of diabetic rats. METHODS: pCMV.Ins, an expression plasmid of the human insulin gene, wrapped with chitosan nanoparticles, was transfected to the diabetic rats through lavage and coloclysis, respectively. Fasting blood glucose and plasma insulin levels were measured for 7 d. Reverse transcription polymerase chain reaction (RT-PCR) analysis and Western blot analysis were performed to confirm the expression of human insulin gene. RESULTS: Compared with the control group, the fasting blood glucose levels in the lavage and coloclysis groups were decreased significantly in 4 d (5.63 ± 0.48 mmol/L and 5.07 ± 0.37 mmol/L vs 22.12 ± 1.31 mmol/L, respectively, P < 0.01), while the plasma insulin levels were much higher (32.26 ± 1.81 μIU/mL and 32.79 ± 1.84 μIU/mL vs 14.23 ± 1.38 μIU/mL, respectively, P < 0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and coloclysis groups. CONCLUSION: Human insulin gene wrapped with chitosan nanoparticles can be successfully transfected to rats through gastrointestinal tract, indicating that chitosan is a promising non-viral vector.
AIM： To study the expression of human insulin gene in gastrointestinal tracts of diabetic rats. METHODS： pCMV.Ins, an expression plasmid of the human insulin gene, wrapped with chitosan nanoparticles, was transfected to the diabetic rats through lavage and coloclysis, respectively. Fasting blood glucose and plasma insulin levels were measured for 7 d. Reverse transcription polymerase chain reaction （RT-PCR） analysis and Western blot analysis were performed to confirm the expression of human insulin gene. RESULTS： Compared with the control group, the fasting blood glucose levels in the lavage and coloclysis groups were decreased significantly in 4 d （5.63±0.48 mmol/L and 5.07±0.37 mmol/L vs 22.12±1.31 mmol/L, respectively, P 〈 0.01）, while the plasma insulin levels were much higher （32.26±1.81μIU/mL and 32.79±1.84μIU/mL vs 14.23±1.38μIU/mL, respectively, P 〈 0.01）. The human insulin gene mRNA and human insulin were only detected in the lavage and coloclysis groups. CONCLUSION： Human insulin gene wrapped with chitosan nanoparticles can be successfully transfected to rats through gastrointestinal tract, indicating that chitosan is a promising non-viral vector.
关键词
1型糖尿病
基因治疗
基因表达
壳聚糖纳米颗粒
人胰岛素基因
Gastrointestinal tract
Human insulin gene
Gene expression
Diabetes mellitus
Chitosan nanoparticle
