Application of intervening in NLRP3 inflammasome in preparation of medicine for treating novel coronavirus pneumonia
A coronavirus and inflammasome technology, applied in antiviral agents, drug combinations, pharmaceutical formulations, etc., can solve the problems of lack of early and effective drug treatment, and achieve the effect of slowing down the storm
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Embodiment 1
[0019] Example 1 Research on the influence of novel coronavirus structural proteins on inflammasomes
[0020] 293T cells were transfected with plasmids S, E, M, N expressing different SARS-CoV-2 proteins. Collect the cell lysate and use Flag beads to co-incubate with the cell lysate, incubate overnight for four times, wash off, and perform detection and analysis by WB. The results are as follows: figure 1 As shown in A, figure 1 The results of A show that the new coronavirus M protein has a direct interaction with NLRP3, and other structural proteins have no direct interaction with NLRP3.
[0021] A549 cells were transfected with a plasmid expressing the new crown protein M, and Flag beads were used to co-incubate with cell lysate, incubated overnight for four times, eluted, and analyzed by WB. The results are as follows figure 1 As shown in B, figure 1 The results of B show that the new crown M protein expressed in A549 cells can interact with NLRP3.
[0022] 293T cells w...
Embodiment 2
[0026] Example 2 Effect of NLRP3 inhibitor MCC950 on activation of NLRP3 inflammasome by M protein of new coronavirus
[0027] THP1 cells were transfected with a plasmid expressing the M protein of the new coronavirus, and MCC950 inhibitors were added at the same time. Cell lysates and cell culture supernatants were collected, and the expression of different molecules was detected by WB. The results were as follows: figure 2 As shown in A, figure 2 A The results showed that the MCC950 inhibitor inhibited the NLRP3 inflammasome activated by the new crown M protein, and inhibited the maturation and release of the cytokine IL-1β and cell pyroptosis.
[0028] THP1 cells were transfected with a plasmid expressing the M protein of the new coronavirus, and MCC950 inhibitors were added at the same time. Collect the cell culture supernatant, and the kit detects the release of lactate dehydrogenase, and the results are as follows: figure 2 As shown in B, figure 2 The results of ...
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