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Acta Physiologica 2011; Volume 202, Supplement 684
The Joint Conference (FAMÉ 2011) of the LXXVth Meeting of the Hungarian Physiological Society, XVIth Meeting of the Hungarian Society of Anatomists, Experimental Section of the Hungarian Society for Experimental and Clinical Pharmacology and Hungarian Society for Microcirculation and Vascular Biology
6/8/2011-6/11/2011
Pécs, Hungary
IN VIVO APPLICATION OF RNAI AS ANTI-OXIDANT THERAPY IN MOUSE ISCHEMIA-REPERFUSION INJURY
Abstract number: O49
Revesz1 Cs., Kaucsar1 T., Racz1 Zs., Hamar1 P.
Aims:
Acute renal failure is a leading cause of death at the intensive care unit and develops frequently due to ischemic acute tubular necrosis (ATN). Post-ischemic apoptosis in the reperfusion phase further aggravates the extent of the injury. Reactive oxygen species (ROS) generated during reoxygenation has a major importance in the apoptotic process. Therefore inhibition of oxidases in ischemia-reperfusion injury (IRI) could have a therapeutic potential. In our study we used RNA interference (RNAi) based post transcriptional gene silencing (PTGS) to inhibit the expression of oxidases in vivo, possibly involved in the development of ATN during renal ischemia-reperfusion injury.
Methods:
Two days before left renal IRI, C57BL/6 mice were injected with 50 mg lipofectamine (10%) mixed siRNA against: NADPH oxidase 2 and 4 (Nox2, Nox4) and xanthine dehydrogenase (Xdh) mRNA, via the left renal vein. After 24 hours of reperfusion blood urea level (Reflotron) ATN (histology), Nox2, Nox4, Xdh and Ngal mRNA (real-time PCR) expression were evaluated.
Results:
In the IRI group blood urea, Ngal and Xdh mRNA levels and ATN score increased significantly, compared to sham operated group. Silencing efficiency was 43.76% and 55.00% following Nox4 and Xdh siRNA treatment, respectively. Surprisingly neither treatment had a significant effect on reperfusion injury.
Conclusion:
We efficiently silenced oxidases in the mouse IRI model, however we observed no protection. A possible explanation could lay in the physiologic redox signaling role of these oxidases.
Support:
OTKA:K81972; ETT: 011-07/2009
| Groups | urea (mg/dl) | NGAL expr | ATN score |
|---|
| Median | 25% | 75% | Median | 25% | 75% | Median | 25% | 75% | |
|---|
| sham | 64.40 | 58.7 | 79.8 | 0.02 | 0.01 | 0.03 | 1.50 | 1.20 | 1.80 |
| IRI | 344.80 | 309.0 | 500.0 | 2.77 | 1.91 | 3.53 | 2.90 | 2.60 | 3.00 |
| NOX2 | 307.50 | 291.0 | 327.0 | 2.64 | 1.61 | 3.48 | 2.95 | 2.89 | 3.00 |
| NOX4 | 321.00 | 293.1 | 327.0 | 2.83 | 1.91 | 3.48 | 2.78 | 2.56 | 3.00 |
| XDH | 319.50 | 286.5 | 336.0 | 1.73 | 1.21 | 2.85 | 2.83 | 2.67 | 3.00 |
| NOX2+4 | 327.00 | 296.7 | 342.0 | 2.58 | 1.95 | 3.66 | 2.90 | 2.88 | 3.00 |
| XDH+NOX2 | 289.65 | 281.4 | 333.2 | 1.90 | 1.43 | 2.43 | 2.68 | 2.50 | 2.90 |
| XDH+NOX4 | 348.00 | 290.1 | 369.0 | 2.02 | 1.16 | 2.93 | 3.00 | 3.00 | 3.00 |
To cite this abstract, please use the following information:
Acta Physiologica 2011; Volume 202, Supplement 684 :O49