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Title:: PDGF-induced decrease of RhoA activity detected by fluorescence resonance energy transfer in airway smooth muscle cells

Author(s):: LI Hui; GUO Jia; DENG Linhong; OUYANG Mingxing; (Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineening & School of Pharmacy，Changzhou University, Changzhou 213164, China)

Keywords:: RhoA; Fluorescence resonance energy transfer; Platelet-derived growth factor; Rac; Asthma

摘要:: Ras同源家族成员A（ras homologous family member A, RhoA）被认为是治疗哮喘的新靶点之一，为探究RhoA在哮喘中的生理机制，本研究利用荧光共振能量转移探针技术实时监测细胞内RhoA活性，发现在血小板衍生生长因子(platelet-derived growth factor,PDGF)的刺激下，气道平滑肌（airway smooth muscle,ASM）细胞中的RhoA信号呈下降趋势。考虑到Rac和RhoA活性存在拮抗效应，本研究进一步抑制细胞中的Rac信号，以及通过转染Vav2激活Rac后，RhoA的下降趋势未受到明显影响；抑制或激活细胞中Cdc42也呈现出类似结果。研究结果表明，PDGF可促进RhoA活性的下调，其不直接受Rac或Cdc42的影响，具体的分子机制仍有待进一步探索。

Abstract:: Ras homologous family member A (RhoA) is recognized as one of the new targets for treatment of asthma. To explore the physiological mechanism of RhoA in asthma, we utilized fluorescence resonance energy transfer probe technology to monitor intracellular RhoA activity in real time.It was found that RhoA signal in airway smooth muscle (ASM) cells showed a decreasing trend under the stimulation of platelet-derived growth factor(PDGF).Considering the antagonistic effect of Rac and RhoA activity, we further inhibited Rac signal in cells, activated Rac by transfection with Vav2. The decline of RhoA was not significantly affected.The inhibition or activation of Cdc42 in cells also showed similar results. The results indicate that PDGF can promote the decrease of RhoA activity in ASM cells, which isn′t directly mediated by Rac or Cdc42, and the molecular mechanism is to be further explored.?