Evaluation Effectiveness of in Vitro Cultivation of Bezoar on Mouse Model Combining Disease with Syndrome of Coronavirus Pneumonia with Yidu Xifei Syndrome / 中国实验方剂学杂志

ABSTRACT

Objective:To determine the therapeutic effect of <italic>in vitro</italic> cultivation of bezoar on a mouse model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome. Method:BALB/c mice were randomly divided into six groups according to their weight grade： normal group， HCoV-229E infection group， cold and damp group， a mouse model combining disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome， and high and low dose group of <italic>in vitro</italic> cultivation of bezoar. The combination model of human coronavirus pneumonia with Yidu Xifei syndrome mice was established by the method of cold dampness condition stimulation+coronavirus HCoV-229E infection. <italic>In vitro</italic> cultivation of bezoar （0.128，0.064 g·kg^-1） was administrated by gavage for 3 days from the day of infection. The observation indexes included： general state observation of mice， inhibition rate of lung index and lung index of mice. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the viral load in the lung tissues of mice. Serum levels of motilin（MTL）， gastrin （GAS）， and cytokines interleukin（IL）-10，IL-6， tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）and interferon-<italic>γ</italic>（IFN-<italic>γ</italic>） in lung tissue of mice were determined by enzyme-linked immunosorbent assay（ELISA）. The percentages of CD4⁺ T lymphocytes，CD8⁺ T lymphocytes and B lymphocytes in the blood of mice were determined by flow cytometry. Result:The high and low dose group of <italic>in vitro</italic> cultivation of bezoar can significantly improve the general condition of model mice. Compared with blank group， model group mice lung index increased significantly （<italic>P</italic><0.01）， nucleic acids significantly increased expression of lung tissue in mice （<italic>P</italic><0.01）， significantly higher serum MTL content in mice， GAS content significantly decreased （<italic>P</italic><0.05，<italic>P</italic><0.01）， lung tissue cells in the immune factor TNF-<italic>α</italic>， IL-10 and IL-6 were significantly increased （<italic>P</italic><0.01）， peripheral blood lymphocyte CD4⁺ T cells in mice， The percentages of CD8⁺ T cells and B cells were significantly decreased （<italic>P</italic><0.01）. Compared with model group， <italic>in vitro</italic> cultivation bezoar mice lung index of high and low dose group were significantly lower （<italic>P</italic><0.01）， the lung tissue of mice express nucleic acid decreased significantly （<italic>P</italic><0.01）， MTL content decreased significantly （<italic>P</italic><0.01）， the lung tissue of mice in the IL-6， IL-10， the TNF-<italic>α</italic>， IFN-<italic>γ</italic> levels were significantly lower （<italic>P</italic><0.01）， <italic>in vitro</italic> cultivation bezoar high dose group can significantly increase the CD4⁺ T cell percentage （<italic>P</italic><0.05）， <italic>in vitro</italic> cultivation bezoar can to a certain extent reduce model mice lung inflammatory exudation， pulmonary interstitial edema， as well as blood stasis symptoms. Conclusion:<italic>In vitro</italic> cultivation of bezoar has a significant therapeutic effect on a mice model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome. It can be treated by reducing the lung index of the model mice， improving the pathological damage of the lung tissue， adjusting the immune effective and inhibiting the clearing of inflammatory factors， and to provide a laboratory basis for clinical medication.