Department of Pharmacology, School of Medicine, Pusan National University, Yangsan, Gyeongnam, Republic of Korea.
Gene & Cell Therapy Research Center for Vessel-associated Diseases, Pusan National University, Yangsan, Gyeongnam, Republic of Korea.
PLoS One. 2022 Mar 16;17(3):e0265191. doi: 10.1371/journal.pone.0265191. eCollection 2022.
Mechanically stressed vascular smooth muscle cells (VSMCs) have potential roles in the development of vascular complications. However, the underlying mechanisms are unclear. Using VSMCs cultured from rat thoracic aorta explants, we investigated the effects of mechanical stretch (MS) on the cellular secretion of high mobility group box 1 (HMGB1), a major damage-associated molecular pattern that mediates vascular complications in stressed vasculature. Enzyme-linked immunosorbent assay (ELISA) demonstrated an increase in the secretion of HMGB1 in VSMCs stimulated with MS (0-3% strain, 60 cycles/min), and this secretion was markedly and time-dependently increased at 3% MS. The increased secretion of HMGB1 at 3% MS was accompanied by an increased cytosolic translocation of nuclear HMGB1; the acetylated and phosphorylated forms of this protein were significantly increased. Among various inhibitors of membrane receptors mediating mechanical signals, AG1295 (a platelet-derived growth factor receptor (PDGFR) inhibitor) attenuated MS-induced HMGB1 secretion. Inhibitors of other receptors, including epidermal growth factor, insulin-like growth factor, and fibroblast growth factor receptors, did not inhibit this secretion. Additionally, MS-induced HMGB1 secretion was markedly attenuated in PDGFR-β-deficient cells but not in cells transfected with PDGFR-α siRNA. Likewise, PDGF-DD, but not PDGF-AA, directly increased HMGB1 secretion in VSMCs, indicating a pivotal role of PDGFR-β signaling in the secretion of this protein in VSMCs. Thus, targeting PDGFR-β-mediated secretion of HMGB1 in VSMCs might be a promising therapeutic strategy for vascular complications associated with hypertension.
机械应激的血管平滑肌细胞(VSMCs)在血管并发症的发展中可能具有重要作用。然而,其潜在机制尚不清楚。本研究使用大鼠胸主动脉组织块培养的 VSMCs,研究了机械拉伸(MS)对细胞分泌高迁移率族蛋白 B1(HMGB1)的影响,HMGB1 是一种主要的损伤相关分子模式,可介导应激血管中的血管并发症。酶联免疫吸附试验(ELISA)显示,MS 刺激的 VSMCs 中 HMGB1 的分泌增加(0-3%应变,60 个周期/分钟),并且在 3% MS 下这种分泌明显且呈时间依赖性增加。3% MS 下 HMGB1 分泌的增加伴随着核 HMGB1 细胞质易位的增加;该蛋白的乙酰化和磷酸化形式明显增加。在介导机械信号的各种膜受体抑制剂中,AG1295(血小板衍生生长因子受体(PDGFR)抑制剂)减弱了 MS 诱导的 HMGB1 分泌。其他受体的抑制剂,包括表皮生长因子、胰岛素样生长因子和成纤维细胞生长因子受体,并没有抑制这种分泌。此外,PDGFR-β 缺陷细胞中 MS 诱导的 HMGB1 分泌明显减弱,但 PDGFR-α siRNA 转染的细胞则不然。同样,PDGF-DD 而非 PDGF-AA 直接增加 VSMCs 中 HMGB1 的分泌,表明 PDGFR-β 信号在 VSMCs 中该蛋白分泌中起关键作用。因此,靶向 PDGFR-β 介导的 VSMCs 中 HMGB1 的分泌可能是治疗与高血压相关的血管并发症的一种有前途的治疗策略。