Application of melatonin in preparation of medicine for inhibiting expression of novel coronavirus SARS-CoV-2 receptor
A coronavirus, sars-cov-2 technology, applied in the medical application field of airway inflammation, to achieve the effect of broadening the types of drugs and reducing allergic airway inflammation
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Embodiment 1
[0022] Example 1: A mouse model of allergic airway inflammation was constructed to detect the regulatory effect of melatonin on allergic airway inflammatory diseases.
[0023] The model of allergic airway inflammation was constructed by using female C57BL / 6 mice aged 6-8 weeks, sensitized on the 0th day and the 7th day, and given intraperitoneal injection of 500 μl ovalbumin (Ovalbumin, OVA) suspension (containing 10 μg OVA and 1 mg potassium aluminum sulfate). The challenge began on the 14th day, and the mice in the model group were given 1% OVA nebulization for 30 minutes, and the nebulization continued for 7 days. Mice in the melatonin intervention group were intraperitoneally injected with 30 mg / kg melatonin 1 hour before each challenge. Tested 24 hours after the last nebulization, if figure 1 Shown in A. After the blood was taken from the mice in each group, the right lung was ligated, and the left lung was lavaged five times with pre-cooled phosphate buffered saline s...
Embodiment 2
[0027] Example 2: In a mouse model of allergic airway inflammation, melatonin can effectively inhibit the expression of OVA-induced SARS-CoV-2-related receptors (ACE2, TMPRSS2, Calmodulin and ADAM17).
[0028]SARS-CoV-2 relies on two key proteins, ACE2 and TMPRSS2, for efficient entry into host cells. The ectodomain of ACE2 can be cleaved into a soluble form by ADAM17, thereby facilitating the entry of SARS-CoV into cells through endocytosis. On the contrary, Calmodulin can inhibit the shedding of ACE2 ectodomain, thereby retaining ACE2 in the cell membrane, and membrane-bound ACE2 can promote SARS-CoV to enter cells through fusion.
[0029] according to figure 1 A mouse model of allergic airway inflammation was constructed, and melatonin was given to intervene. The expression of SARS-CoV-2 receptors ACE2, TMPRSS2, Calmodulin and ADAM17 in mouse lung tissue was detected by western blot. The results showed that OVA stimulation significantly increased the protein expression o...
Embodiment 3
[0032] Example 3: Melatonin significantly inhibited the expression of inflammatory factors in human primary bronchial epithelial cells (HBECs) after OVA stimulation.
[0033] To examine the regulatory effect of melatonin on OVA-induced inflammation in human bronchial epithelial cells, primary human bronchial epithelial cells (HBECs), which are the main target cells of SARS-CoV-2, were isolated and cultured in this example. The cells were obtained from lung cancer patients undergoing lobectomy, and bronchial epithelial cells were isolated from the airway tissue >3cm from the edge of the tumor tissue, and cultured at 37°C with 5% CO 2 condition.
[0034] The identification of primary human bronchial epithelial cells was labeled with cytokeratin CK17 / 19 by immunofluorescence, and the cells in the logarithmic growth phase were taken to prepare cell slides, which were fixed, permeabilized, and blocked, then incubated with CK17 / 19 antibodies, and passed overnight at 4°C , immunoflu...
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