Application of nitrogen-containing bisphosphonate in prevention or treatment of viral pneumonia
A technology for viral pneumonia and bisphosphonates, which can be used in biological, clinical, and medical fields to solve problems such as the effects of bisphosphonates that have not yet been discovered
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Embodiment 1
[0092] Example 1. Alendronate sodium can promote macrophages to eliminate SARS-CoV-2Delta strains in vitro
[0093] 1. Experimental steps
[0094] Take mouse alveolar macrophages, according to 5×10 per well 4 cells were plated in 24-well plates. The next day follow 2.5×10 4 After 2 hours of TCID50 virus infection, the culture medium was discarded, washed twice with PBS, and the PBS was discarded for the last time. Add 1mL of 50μM alendronate sodium to each well, continue to culture for 22h or 46h, then wash twice with PBS, and discard the PBS for the last time. Add 1mL TRIZOL to each well, extract RNA, and perform reverse transcription. Analysis of viral gene ORF1ab( Figure 1A ) and N( Figure 1B ) copy number.
[0095] 2. Experimental results
[0096] qPCR analysis showed that alendronate sodium can reduce the viral load of SARS-CoV-2Delta strain in macrophages in vitro ( Figure 1A with Figure 1B ).
Embodiment 2
[0097] Example 2. Alendronate sodium can down-regulate the expression of IL-6 in macrophages infected with Delta strain in vitro
[0098] 1. Experimental steps
[0099] Take mouse alveolar macrophages, according to 5×10 per well 4 cells were plated in 24-well plates. The next day follow 2.5×10 4 After 2 hours of TCID50 virus infection, the culture medium was discarded, washed twice with PBS, and the PBS was discarded for the last time. Add 1mL of 50μM alendronate sodium to each well, continue to culture for 22h or 46h, then wash twice with PBS, and discard the PBS for the last time. Add 1mL TRIZOL to each well, extract RNA, and perform reverse transcription. The expression of IL-6 was analyzed by qPCR.
[0100] 2. Experimental results
[0101] Alendronate sodium can down-regulate the expression of IL-6 in macrophages infected with Delta strain in vitro ( figure 2 ).
Embodiment 3
[0102] Example 3. Alendronate sodium can increase the endosomal pH value of macrophages both in vivo and in vitro
[0103] 1. Experimental steps
[0104] In vitro pH measurement: take mouse alveolar macrophages, according to 5×10 per well 4 The cells were plated into confocal small dishes, and then 1 mL of 50 μM alendronate sodium was added to each well, and cultured for 24 hours. Then add pHrodo TM Red dextran was labeled for 10 min and photographed with a confocal microscope.
[0105] In vivo pH measurement: intranasally administered 150 μg alendronate sodium to each mouse, 24 hours later, the alveolar macrophages of the mice were taken out, and 50 μg / mL pHrodo TM Red dextran labeled for 10min with NxT Acoustic Focusing Flow Cytometry Detection.
[0106] 2. Experimental results
[0107] Alendronate sodium treatment of mouse alveolar macrophages in vitro can increase the pH value of the endosome ( Figure 3A , Figure 3B ). In vivo administration of alendronate sodi...
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