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1 Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh School of Medicine and Magee-Womens Research Institute, Pittsburgh, Pennsylvania, USA
2 Department of Zoology and Howard Florey Institute, University of Melbourne, Parkville, Victoria, Australia
3 Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh School of Medicine and Magee-Womens Research Institute, Pittsburgh, Pennsylvania, USA; Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
* To whom correspondence should be addressed. E-mail: rsikpc{at}mwri.magee.edu.
Vascular gelatinase activity is essential for pregnancy and relaxin-induced renal vasodilation and hyperfiltration in rats. The objective of this study was to further elucidate the mechanisms for the increase in vascular MMP-2 activity caused by pregnancy and relaxin. We first corroborated our earlier work by showing that both pro and active MMP-2 were increased in small renal arteries from pregnant compared to virgin rats and relaxin-treated compared to vehicle-treated nonpregnant rats. We next investigated other artery types, and showed that MMP-2 activity was upregulated in mesenteric arteries from pregnant rats (pro MMP-2 by 50% and active MMP-2 by 40%, both p<0.05) and from relaxin-treated nonpregnant rats (pro MMP-2 by 50% and active MMP-2 by 90%, both p<0.005) compared to their respective controls. To corroborate these results obtained by gelatin zymography, pro MMP-2 protein was determined by Western analysis in the same small arteries. Pro MMP-2 protein was increased in small renal arteries from pregnant compared to virgin rats (pro MMP-2/
-actin: 0.29 vs 0.21, p<0.01) and from relaxin-treated compared to vehicle-treated nonpregnant rats (0.43 vs 0.32, p<0.005). Similar findings were observed for mesenteric arteries. MMP-2 mRNA as measured by real time PCR was increased in small renal arteries from pregnant and relaxin-treated nonpregnant rats compared with their respective controls. There were no significant differences in TIMP-1 or TIMP-2 activity by reverse zymography in small renal arteries. Thus, increases in MMP-2 mRNA and protein expression are major factors contributing to increased MMP-2 activity in small arteries from pregnant and relaxin-treated nonpregnant rats. of small renal arteries isolated from rats.
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