Found programs:
Authors:Gong Caiwei; Zhao Guangjian; Liu Danan; Ou Hangjun; Zhao Quanwei; Li Hui
Keywords:dapagliflozin;human myeloid leukemia monocytes;foam cells;pyroptosis;oxidized low-density lipoprotein
DOI:10.19405/j.cnki.issn1000-1492.2023.08.020
〔Abstract〕 Objective To investigate the role of dapagliflozin(DAPA) in ox-LDL-induced pyroptosis of human myeloid leukemia monocytes(THP-1) derived foam cells. Methods THP-1-derived foam cell pyroptosis model was constructed by ox-LDL-induced THP-1derived macrophages. The experimental groups were set as follows: the blank control group(NC), the ox-LDL group(ox-LDL), and the drug intervention group(ox-LDL+DAPA). Oil Red O staining was used to detect the foam cell levels of macrophages. The cell proliferation and toxicity assay kit was used to detect the effect of DAPA on foam cell viability. Hoechst 33342/propidium iodide(PI) double staining was used to detect THP-1 derived foam cell pyroptosis. Cell immunofluorescence double staining was used to detect the effect of DAPA on the expression of pyroptosis key factor Caspase-1 in foam cells. The activity of lactate dehydrogenase(LDH) in the cell culture medium was detected using a microplate enzyme-linked immunosorbent assay. qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of Nod-like receptor pyrin domain containing 3(NLRP3), cystein-containing aspartate-specific protease-1(Caspase-1),apoptosis-associatedspeck-like protein containing CARD(ASC),gasdermin-D(GSDMD), interleukin(IL)-18andIL-1β,respectively. Results The CCK-8 assay indicated that the optimal intervention concentration of DAPA was 10 μmol/L. Oil Red O staining confirmed the successful construction of the THP-1-derived foam cell pyroptosis model. Compared with the blank control group, the expression levels ofNLRP3,Caspase-1,ASC,GSDMD,IL-18,IL-1βmRNA and protein significantly increased in ox-LDL group(P<0.05), as well as the number of PI-positive cells and LDH activity(P<0.05), the fluorescence intensity of Caspase-1 and the number of redlipid droplets in the cytoplasm of the cells. However, these effects were significantly reversed after DAPA intervention in the ox-LDL+DAPA group(P<0.05). Conclusion DAPA inhibits ox-LDL-induced pyroptosis in THP-1-derived foam cells.