The role of dapagliflozin in ox⁃LDL⁃triggered pyroptosis of THP⁃1 ⁃derived foam cells
10.19405/j.cnki.issn1000-1492.2023.08.020
- Author:
Caiwei Gong
1
,
2
;
Guangjian Zhao
1
,
2
;
Danan Liu
1
,
2
;
Hangjun Ou
1
,
2
;
Quanwei Zhao
1
,
2
;
Hui Li
1
,
2
Author Information
1. Dept of Cardiology , Afiliated Hospital of Guizhou Medical University, Guiyang 550004
2. Institute of Medical Sciences , Guizhou Medical University , Guiyang 550004
- Publication Type:Journal Article
- Keywords:
dapagliflozin;
human myeloid leukemia monocytes;
foam cells;
pyroptosis;
oxidized low⁃density lipoprotein
- From:
Acta Universitatis Medicinalis Anhui
2023;58(8):1366-1373
- CountryChina
- Language:Chinese
-
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 Ostaining 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 pyrindomain containing 3 (NLRP3) , cystein⁃containing aspartate⁃specific protease⁃1( Caspase⁃1 ) , apoptosis⁃associated⁃speck⁃like protein containing CARD(ASC) ,gasdermin⁃D (GSDMD) , interleukin(IL) Ⅳ18 and IL⁃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 of NLRP3 , Caspase⁃1 , ASC , GSDMD , IL⁃18 , IL⁃1β mRNA 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 .
- Full text:2024081909524848269达格列净在ox-LDL诱导...1源性泡沫细胞焦亡中的作用_龚才伟.pdf
