Mechanical Stretch Enhances mRNA Expression and Proenzyme Release of Matrix Metalloproteinase-2 (MMP-2) via NAD(P)H Oxidase–Derived Reactive Oxygen Species

Abstract

Mechanical stretch is a hallmark of arterial hypertension and leads to vessel wall remodeling, which involves matrix metalloproteinases (MMPs). Because mechanical stretch is further capable of inducing reactive oxygen species (ROS) formation via the NAD(P)H oxidase, we assessed whether mechanical stretch enhances MMP expression and activity in a NAD(P)H oxidase-dependent manner. Therefore, vascular smooth muscle cells (VSMCs) isolated from C57BL/6 mice were exposed to cyclic mechanical stretch. The impact of ROS was assessed using VSMCs isolated from p47phox-/- mice, deficient for a NAD(P)H oxidase subunit responsible for ROS formation. Transcript levels were investigated by cDNA array and confirmed by RT-PCR. ROS formation was determined by DCF fluoroscopy and MMP-2 activity by zymography. Mechanical stretch of wild-type VSMCs resulted in a rapid ROS formation and p47phox membrane translocation that is followed by an increase in Nox-1 transcripts. ROS formation was completely abrogated in p47phox-/- VSMCs. cDNA array further revealed an increase of MMP-2 mRNA in response to mechanical stretch, which was validated by RT-PCR. Using p47phox-/- VSMCs, this increase in MMP-2 mRNA was completely blunted. mRNA expression of tissue inhibitor of MMP-2 TIMP-1 and TIMP-2 and membrane-type 1 MMP was unaffected by mechanical stretch. Gelatinolytic activity of pro-MMP-2 has been increased rapidly in wild-type VSMCs and was completely abolished in p47phox-/- VSMCs. These results indicate that mechanical stretch induces ROS formation via the NAD(P)H oxidase and thereby enhances MMP-2 mRNA expression and pro-MMP-2 release. These results are consistent with the notion that in arterial hypertension, reactive oxygen species are involved in vascular remodeling via MMP activation. The full text of this article is available online at http://www.circresaha.org.

Stretch-inducible Expression of the Angiogenic Factor CCN1 in Vascular Smooth Muscle Cells Is Mediated by Egr-1

“…Furthermore, Yamaguchi et al (30) reported that stretch-dependent Egr-1 activation enhances the expression of membrane type 1 matrix metalloproteinase in rat microvascular endothelial cells with a significant impact on angiogenesis by affecting the activity of matrix metalloproteinase-2. In this regard, we recently demonstrated that mechanical stretch enhances expression and release of matrix metalloproteinase-2 in a redox-sensitive manner (11).…”

Redox Control of Renal Function and Hypertension

“…Furthermore, the prominent increase of p47 phox mRNA and protein expression observed in 4-week-old SHRs in the vasculature, macula densa, and distal nephron demonstrates that oxidative stress precedes development of hypertension. Mechanical stretch of wild-type VSMCs resulted in ROS formation and p47 phox translocation to the plasma membrane followed by an increase in Nox1 transcripts (92). In addition, Ang II-stimulated p47 phox translocation through c-Src phosphorylation and enhanced activity of NAD(P)H oxidase in human VSMCs (328).…”

Functions of epidermal growth factor receptor in cisplatin response of thyroid cells

“…But the signalling mechanisms by which cisplatin stimulation modulated MMP-2 expression in thyroid cells are not yet clear. Several studies suggest that reactive oxygen species (ROS) play a major role in the activation of MMPs [57,58], and that NAD(P)H oxidase activation, a major source of ROS, is a key event in this process [59]. We here show that the inhibition of NAD(P)H oxidase not only prevented cisplatin-induced ROS generation and increase of MMP-2, but also inhibited p38/MAPK and EGFR activation, caspase-7 proteolysis and the arise of the apoptotic intrinsic pathway (Fig.…”