Script com entonacao - Juliane

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<p>Hi, my name is Juliane, I'm a master's student at UNESP Araçatuba, my paper is about the test of actions of the SARS-CoV-2 spike protein and angiotensin 2 in human bronchial smooth muscle cells.</p><p>SARS-CoV-2 is the virus that causes the covid 19 disease, it is known that this virus is composed of 4 main structural proteins, M-membrane, E-envelope, N-nuclecapsid and S-spike. The protein is responsible for the internalization of the virus into the host cell, due to its connection with ACE2 present on the surface of these cells.</p><p>ACE2 is an important component of RAS, this enzyme converts Ang2 into Ang1-7 which acts on its MAS receptor promoting anti-inflammatory actions.</p><p>The objectives of this paper was to investigate whether a spike protein stimulates the expression of ACE, ACE2 and MasR mRNA and the production of the cytokine IFN-gamma in these smooth muscle cells.</p><p>A primary human bronchial smooth muscle cell was used and kept in culture in a controlled environment at 37° at 5% CO2 until reaching the appropriate confluence for the experiment. It is then plated in a 24-well plate at a concentration of 50 thousand cells per well, after 24 hours of plating, these cells begin to be stimulated with the spike protein at 3 different concentrations (1 ug/mL, 5 ug/mL and 10 ug/mL) and also with Ang2. The experiment happens in 3 sceneries: i) a group of cells reacting only with Ang2. ii) a second group reacting with Ang2 and the S protein added at the same time and iii) a third group when we added the Ang2 3 hours after the addition of S protein. When isolated the stimulus occurs 3 hours after the first stimulus with the S protein. These stimulus remain for periods of 3, 24 and 48 hours, and at each time a sample was collected for analysis, supernatant for ELISA and RNA removal with trizol kit for qRT-PCR</p><p>In the results we notice a reduction in gene expression of ACE, ACE2 and MasR when cells were stimulated with a spike protein. There was a reduction in ACE at a concentration of 5 ug/mL at 3 and 24 hours. In our tests, we could see that stimulations with Ang2 seems to reverse the inhibition when we added the S protein hours after. ACE2 presents the same profile, there is a reduction in ACE2 gene expression when stimulated with spike protein within 3 hours, but stimulation with Ang2 added at the same time appears to increase this gene expression within 24 hours. For the MAS receiver the same settings are displayed for 24- and 48-hour period.</p><p>The production of IFN-gamma was increased by the spike protein at a concentration of 10 ug/mL at 3 and 24 hours, Ang2 added after 3 hours appears to model this expression at 24 hours. The 5 ug/mL spike protein decreases the production of IFN-gamma within 3 hours and Ang2 added after 3 hours reverses this inhibition.</p><p>So far, we can conclude that the spike protein inhibits the gene expression of ACE, ACE2 and MasR and the production of IFN-gamma, and Ang2 appears to modulate this.</p><p>Inváiroument</p><p>édidi</p>