Table 2.
Treatment
|
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Serum Parameter | Saline Only | PP | MC | MC & Stuffer DNA | |
AST (IU/L) | 2 weeks before injection | 105 ± 64 | 98 ± 4 | 68 ± 4 | 68 ± 11 |
1 day after injection | 1810 ± 1598 | 2885 ± 1075 | 1258 ± 4 | 1925 ± 304 | |
2 weeks after injection | 88 ± 25 | 85 ± 21 | 73 ± 11 | 83 ± 18 | |
ALT (IU/L) | 2 weeks before injection | 58 ± 18 | 78 ± 32 | 40 ± 7 | 63 ± 4 |
1 day after injection | 3130 ± 2178 | 4958 ± 2110 | 2240 ± 976 | 3195 ± 382 | |
2 weeks after injection | 65 ± 21 | 48 ± 11 | 18 ± 4 | 50 ± 14 | |
IL-12 p40 (pg/ml) | 2 weeks before injection | 96 ± 11 | 134 ± 58 | 210 ± 71 | 130 ± 23 |
1 day after injection | 68 ± 8 | 92 ± 47 | 157 ± 45 | 103 ± 36 | |
2 weeks after injection | 103 ± 31 | 133 ± 40 | 186 ± 36 | 139 ± 26 | |
TNF-α (pg/ml) | 2 weeks before injection | 2.2 ± 1.1 | 1.5 ± 0.7 | 1.1 ± 0.1 | 1.4 ± 0.5 |
1 day after injection | 2.3 ± 0.6 | 3.7 ± 1.5 | 1.6 ± 0.9 | 4.6 ± 0.4 | |
2 weeks after injection | 1.6 ± 0.9 | 1.3 ± 0.4 | <1 | 4.4 ± 1.3 | |
IFN-γ (pg/ml) | 2 weeks before injection | <3 | <3 | <3 | <3 |
1 day after injection | <3 | <3 | <3 | <3 | |
2 weeks after injection | <3 | <3 | <3 | <3 |
The concentration of liver specific enzymes including aspartate aminotransferase (AST) and alanine aminotransferase (ALT), as well as the inflammatory biomarkers interleukin-12 (IL-12 p40), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) were assessed 2 weeks before, and 1 day or 2 weeks after hydrodynamic delivery of saline only, parental plasmid (PP) pMC.PKU20, minicircle (MC) MC.PKU20, or minicircle (MC) MC.PKU20 plus stuffer DNA (pMC.BESPX, empty vector). Except for the MC treatment, equal amount of DNA was delivered to the mice, i.e. 219 μg of PP or 77 μg MC or 77 μg MC plus 142 μg stuffer DNA. All vector preparations used were supposed to contain < 0.1 endotoxin units per μg DNA according to the Qiagen endofree plasmid purification kit (Qiagen AG, Hombrechtikon, Switzerland).
AST, aspartate aminotransferase; ALT, alanine aminotransferase; IL-12, interleukin-12 p40; TNF-α, tumor necrosis factor-alpha; IFN-γ, interferon-gamma; 2 mice were measured in each group; data represent mean ± SD.