Gene therapy for type 1 diabetes mellitus in rats by gastrointestinal administration of chitosan nanoparticles containing human insulin gene.
OBJECTIVE
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 microIU/mL and 32.79 +/- 1.84 microIU/mL vs 14.23 +/- 1.38 microIU/mL, respectively, P < 0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and coloclysis groups.


CONCLUSIONS
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.
Niu
Li
L
Department of Endocrinology, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuhan 430071, Hubei Province, China.
Xu
Yan-Cheng
YC
Dai
Zhe
Z
Tang
Hui-Qin
HQ
D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D048271:Chitosan / Q000008:administration & dosage*; D004247:DNA / Q000008:administration & dosage*; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000628:therapy*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007328:Insulin / Q000097:blood / Q000235:genetics*; D008297:Male; D053758:Nanoparticles / Q000008:administration & dosage*; D051381:Rats; D017208:Rats, Wistar
D001786:Blood Glucose; D007328:Insulin; D004247:DNA; D048271:Chitosan