Hydrodynamics-based transfection of pancreatic duodenal homeobox 1 DNA improves hyperglycemia and is associated with limited complications in diabetic mice.
The biohazards caused by the viral delivery of pancreatic duodenal homeobox gene 1 (Pdx1) to the murine liver limits its application. We aimed to evaluate the feasibility of hydrodynamics-based transfection (HBT) with Pdx1 in improving hyperglycemia. Murine hepatocellular carcinoma (Hepa1-6) cells were transfected with the Pdx1-expressing plasmid, pcDNA3.1/V5-His A (pcDNA)-Pdx1. Hepatic delivery of pcDNA-Pdx1 or pcDNA in streptozocin- induced diabetic mice was achieved by HBT. The sequential serum glucose and alanine aminotransferase (ALT) levels were assessed. On the 3(rd) day after transfection, the transfection efficiency in the Hepa1-6 cells and the mice livers was 5% and 0.35 %, respectively. At 1 wk after HBT, asides from hepatic expression of insulin, the diabetic mice transfected with pcDNA-Pdx1 had a significantly lower sugar (211 +/- 61.6 vs. 413 +/- 62 mg/dL; p = 0.002) level than those transfected with pcDNA; however, the difference diminished afterward. No significant difference in the ALT levels was observed between the 2 groups. No mortality was noted in the mice transfected with pcDNA-Pdx1. The hypoglycemic effect of Pdx1 delivered by HBT was transient and associated with negligible complications. In studies on the short-term biological effects of Pdx1 in vivo, HBT is a potential alternative to viral delivery of Pdx1 to the murine liver.
Chen
Tsung-Hsing
TH
Liver Research Center and Department of Hepatogastroenterology, Chang Gung Memorial Hospital, Kuei Shan, Taoyuan, Taiwan.
Yeh
Chau-Ting
CT
Ho
Yu-Pin
YP
Hsu
Chen-Ming
CM
Huang
Chau-Chun
CC
Shiau
Shuen-Shian
SS
Liang
Chun-Kai
CK
Chang
Ming-Ling
ML
D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D045744:Cell Line, Tumor; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000628:therapy*; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006439:Hemodynamics / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006943:Hyperglycemia / Q000517:prevention & control*; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008111:Liver Function Tests; D008297:Male; D051379:Mice; D010179:Pancreas / Q000378:metabolism / Q000473:pathology; D010957:Plasmids; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D001786:Blood Glucose; D018398:Homeodomain Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein