Abstract
The current situation of the SARS-CoV-2 virus infection causing COVID-19 is more serious. The development of antiretroviral drugs is urgently important. To stop the spread of the virus, the lungs model is critical to develop of antiretroviral drugs. Due to, SARS-CoV-2 relies on the Angiotensin-converting enzyme 2 (ACE2) target receptor protein, which is more common in lung cells, and caused of pneumonia. Thus, the focus of this research is to develop a lung model to study the mechanism of viral infection and develop the treatment of COVID-19, because of it is difficult to study biopsy samples from infected people. The aim of this research project is to simulate lung tissue structures using a technique as the Air-liquid interface to create conditions like the pulmonary scaffolds with ACE2 receptor protein expression. Development of therapeutic and drug testing guidelines for COVID-19. The experimental results in two-dimensional A549 lung cancer cells that were induced inflammation by lipopolysaccharide (LPS) was found the highest protein concentration of 0.2 micrograms per microliter. The nitric oxide measurement was observed in LPS-stimulated samples for 24 hours. Inflammation was observed with low nitric oxide about 2 µM, while the three-dimensional with LPS and hydrogen peroxide (H2O2) stimulation, the nitric oxide was not detected. As for the expression of ACE2 receptor that analyzed by Western blot method, the result can be found at approximately 97 kilodalton (kDa) size of the ACE2 receptor. Consequently, the lungs cancer cell line A549 in three-dimensional Air-Liquid Interface was expression ACE-2 receptor. However, the experiments also need to be repeated in the analysis of ACE2 and AT1R expression and control as beta-actin with western blot, and further at the gene level through qPCR analysis in a two-dimensional versus three-dimensional model. To determine the difference in receptor expression in inflammatory stimulation. The model will further investigation of the substance and the stimulation concentration was selected with spike protein instead for stimulation. The antioxidant was measured, and nitrate content will detect inflammation by LPS and H2O2. The problems that cannot trigger inflammation. So, the experiment still needs quality testing and test the concentration of the substance again.
