PMID- 36689357
OWN - NLM
STAT- MEDLINE
DCOM- 20230412
LR  - 20230412
IS  - 1078-6791 (Print)
IS  - 1078-6791 (Linking)
VI  - 29
IP  - 3
DP  - 2023 Apr
TI  - FNDC5/PPARa Pathway Alleviates THP-1-derived Macrophage Pyroptosis and Its 
      Mechanism.
PG  - 32-42
LID - AT7815 [pii]
AB  - CONTEXT: Atherosclerosis (AS) is a chronic inflammatory disease. Pyroptosis is a 
      newly discovered, pro-inflammatory cell death that can trigger and amplify the 
      occurrence and progression of AS. Researchers are still uncertain about the 
      anti-atherosclerotic mechanism of "fibronectin type III domain-containing protein 
      5" (FNDC5). OBJECTIVE: The study aimed to investigate the ability of 
      FNDC5-mediated, "peroxisome proliferator activated receptor alpha" (PPARa) to 
      inhibit oxidized low-density lipoprotein (ox-LDL)-induced, THP-1-derived 
      macrophage pyroptosis and to determine a potential molecular mechanism at the 
      cellular level. DESIGN: The research team performed a laboratory study. SETTING: 
      The study took place in the Department of Cardiovascular Medicine at the 
      Affiliated Hospital of Guzhou Medical University at the Medical Research 
      Institute at Guizhou Medical University in Guiyang, Guizhou, China. OUTCOME 
      MEASURES: The research team: (1) constructed and stably transfected FNDC5 
      gene-overexpressing and FNDC5 gene-silencing lentiviral vectors into THP-1 cells; 
      (2) observed the cell morphology under an inverted fluorescence microscope and 
      screened the stably transfected THP-1 cells with puromycin; (3) verified the 
      transfection efficiency using quantitative real-time polymerase chain reaction 
      (qRT-PCR) and Western Blot; (4) used phorbol to induce THP-1 cells into 
      macrophages; (5) cultured the THP-1-derived macrophages with different 
      concentrations of ox-LDL-25, 50, 75, and 100 microg/ml-for 24 h; (6) performed 
      Hoechst 33342/ propidium iodide (PI) double staining and examined lactate 
      dehydrogenase (LDH) and interleukin-1 beta (IL-1ss) activity to determine the 
      effects of ox-LDL on THP-1-derived macrophage pyroptosis; (7) selected the 
      optimal ox-LDL concentration; (8) divided the THP-1-derived macrophages into 
      seven groups: NC group (no ox-LDL intervention), ox-LDL group, PBS group, Mock1 
      group, Ad-FNDC5 group, Mock2 group, and Sh-FNDC5 group; (9) examined the 
      expressions of functional proteins and the pyroptosis of THP-1-derived 
      macrophages, including FNDC5, PPARa, and "nuclear factor kappa-light chain 
      enhancer of activated B cells P65" (NF-kappaB P65), and those related to the 
      pyroptosis pathway, using Western Blot and Hoechst 33342/PI double staining, 
      respectively; (10) treated the THP-1-derived macrophages with FNDC5 expression 
      with GW6471, a specific PPARalpha antagonist; (11) determined the expressions of 
      functional proteins and the pyroptosis of THP-1-derived macrophages, including 
      FNDC5, PPARa, and NF-kappaB P65, and those related to the pyroptosis pathway, using 
      Western Blot and Hoechst 33342/PI double staining and detection of the LDH and 
      IL-1ss activity, respectively. RESULTS: With the stably transfected THP-1 cells 
      with FNDC5 overexpression or silencing the ox-LDL-induced, THP-1-derived, 
      macrophage pyroptosis occurred in a concentration-dependent manner. Compared with 
      the ox-LDL, phosphate buffered saline (PBS), Mock1, and Mock2 groups, the 
      Ad-FNDC5 group had a significant increase in expression of FNDC5 and of 
      peroxisome proliferator activated receptor alpha (PPARa) proteins (P < .05). The 
      "nuclear factor kappa-light chain enhancer of activated B cells P65: (NF-kappaB P65), 
      NOD-like receptor thermal protein domain associated protein 3, (NLRP3), 
      Caspase-1, gasdermin D (GSDMD, IL-1ss and IL-18 protein expressions, percentage of 
      PI-positive cells, LDH activity, and IL-1ss activity decreased significantly (P < 
      .05); the results in the Sh-FNDC5 group were opposite to those in the Ad-FNDC5 
      group. 3. Intervention with GW6471 (PPARa antagonist) in the stably transfected 
      THP-1-derived macrophages with FNDC5 overexpression abolished the protective 
      effect of FNDC5 against ox-LDL-induced THP-1-derived macrophage pyroptosis. 
      CONCLUSIONS: Irisin/PPARa inhibited THP-1-derived macrophage pyroptosis and 
      inflammation and delayed AS by inhibiting the NF-kappaB/NLRP3 pathway.
FAU - Zhou, Bo
AU  - Zhou B
FAU - Zhao, Guangjian
AU  - Zhao G
FAU - Li, Hui
AU  - Li H
FAU - Zhao, Quanwei
AU  - Zhao Q
FAU - Liu, Danan
AU  - Liu D
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Altern Ther Health Med
JT  - Alternative therapies in health and medicine
JID - 9502013
RN  - 0 (NF-kappa B)
RN  - 0 (PPAR alpha)
RN  - 0 (NLR Family, Pyrin Domain-Containing 3 Protein)
RN  - 0 (Fibronectins)
RN  - 0 (FNDC5 protein, human)
SB  - IM
MH  - Humans
MH  - *NF-kappa B/metabolism/pharmacology
MH  - *PPAR alpha/metabolism/pharmacology
MH  - Pyroptosis
MH  - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism
MH  - Fibronectins/metabolism/pharmacology
MH  - China
MH  - Macrophages/metabolism
EDAT- 2023/01/24 06:00
MHDA- 2023/04/12 06:42
CRDT- 2023/01/23 13:22
PHST- 2023/04/12 06:42 [medline]
PHST- 2023/01/24 06:00 [pubmed]
PHST- 2023/01/23 13:22 [entrez]
AID - AT7815 [pii]
PST - ppublish
SO  - Altern Ther Health Med. 2023 Apr;29(3):32-42.