Human pulmonary microvascular endothelial cells (HPMECs) line blood vessels in the lungs and experience and respond to shear stress. Shear stress is the tangential force of blood flow over the HPMECs. Shear stress is altered in disease which can lead to changes in gene expression from the endothelial cells(1). Changes in gene expression show that HPMECs sense the shear stress that acts upon them. The projects aim is to develop a model to apply shear stress to HPMECs in vitro to better mimic physiological and pathophysiological conditions in comparison to methods previously used. HPMECs will be grown in vitro on six-well plates using the new model statically or in an orbital shaker with high, low, and normal levels of shear stress. In response to shear stress krüppel-like transcription factor 2 (KLF2) and endothelial nitric oxide synthase (eNOS) have been shown to be upregulated(1). To determine which HPMECs closer mimic in vivo conditions KFL2 and eNOS levels will be measured using real time-polymerase chain reaction (RT-PCR). The KFL2 and eNOS levels produced by cells subjected to normal shear stress should be higher in comparison to the cells subjected to high or low shear stress. The control should not express KLF2 and eNOS. KLF2 regulates alterations in gene expression and is crucial for the health of vasculature by providing anti-inflammatory effects(1). eNOS produces nitric oxide which regulates vascular tone and homeostasis(1). KLF2 and eNOS expression from HPMECs which will have undergone normal and abnormal shear stress will be measured, increased expression will suggest that conditions in-vitro better mimic in vivo physiological conditions. By developing a model of applying shear stress to HPMECs in vitro to better mimic physiological conditions in vivo will allow researchers to further investigate shear stress and its impact in diseases such as COPD, pulmonary fibrosis, and pulmonary embolism.
