Department of Physiology, Faculty of Medicine, Ordu University, Ordu, Turkey.
Department of Bioengineering, University of California San Diego, La Jolla, CA, 92093-0412, USA.
Hypertens Res. 2018 Jun;41(6):406-413. doi: 10.1038/s41440-018-0023-7. Epub 2018 Mar 19.
Increased protease activity causes receptor dysfunction due to extracellular cleavage of different membrane receptors in hypertension. The vasodilatory effects of insulin-like growth factor-1 (IGF-1) are decreased in hypertension. Therefore, in the present study the association of an enhanced protease activity and IGF-1 receptor cleavage was investigated using the spontaneously hypertensive rats (SHRs) and their normotensive Wistar Kyoto (WKY) controls (n = 4). Matrix metalloproteinase (MMP) activities were determined using gelatin zymography on plasma and different tissue samples. WKY aorta rings were incubated in WKY or SHR plasma with or without MMP inhibitors, and immunohistochemistry was used to quantify the densities of the alpha and beta IGF-1 receptor (IGF-1R) subunits and to determine receptor cleavage. The pAkt and peNOS levels in the aorta were investigated using immunoblotting as a measure of IGF-IR function. Increased MMP-2 and MMP-9 activities were detected in plasma and peripheral tissues of SHRs. IGF-1R beta labeling was similar in both groups without plasma incubation, but the fraction of immunolabeled area for IGF-1R alpha was lower in the endothelial layer of the SHR aorta (p < 0.05). A 24-h incubation of WKY aorta with SHR plasma did not affect the IGF-1R beta labeling density, but reduced the IGF-1R alpha labeling density in the endothelium (p < 0.05). MMP inhibitors prevented this decrease (p < 0.01). Western blot analyses revealed that the pAkt and peNOS levels under IGF-1-stimulated and -unstimulated conditions were lower in SHRs (p < 0.05). A reduced IGF-1 cellular response in the aorta was associated with the decrease in the IGF-1R alpha subunit in the SHR hypertension model. Our results indicate that MMP-dependent receptor cleavage contributed to the reduced IGF-1 response in SHRs.
在高血压中,由于不同膜受体的细胞外切割,蛋白酶活性的增加导致受体功能障碍。胰岛素样生长因子-1(IGF-1)的血管舒张作用在高血压中降低。因此,在本研究中,使用自发性高血压大鼠(SHR)及其正常血压的 Wistar Kyoto(WKY)对照(n=4)研究了增强的蛋白酶活性和 IGF-1 受体切割的相关性。使用明胶酶谱法在血浆和不同组织样本中测定基质金属蛋白酶(MMP)活性。将 WKY 主动脉环在 WKY 或 SHR 血浆中孵育,有或没有 MMP 抑制剂,并使用免疫组织化学定量测定 alpha 和 beta IGF-1 受体(IGF-1R)亚基的密度,并确定受体切割。使用免疫印迹法研究主动脉中的 pAkt 和 peNOS 水平,作为 IGF-IR 功能的指标。在 SHR 的血浆和外周组织中检测到 MMP-2 和 MMP-9 活性增加。在没有血浆孵育的情况下,两组的 IGF-1R beta 标记相似,但 SHR 主动脉内皮层的 IGF-1R alpha 免疫标记区域分数较低(p<0.05)。用 SHR 血浆孵育 24 小时的 WKY 主动脉不会影响 IGF-1R beta 标记密度,但会降低内皮中的 IGF-1R alpha 标记密度(p<0.05)。MMP 抑制剂可防止这种减少(p<0.01)。Western blot 分析显示,在 SHR 中,IGF-1 刺激和非刺激条件下的 pAkt 和 peNOS 水平较低(p<0.05)。在 SHR 高血压模型中,主动脉中 IGF-1 细胞反应的降低与 IGF-1R alpha 亚基的减少有关。我们的结果表明,MMP 依赖性受体切割导致 SHR 中 IGF-1 反应降低。