Related Experiment Video

Updated: Dec 27, 2025

Evaluation of Caspase Activation to Assess Innate Immune Cell Death
10:23

Evaluation of Caspase Activation to Assess Innate Immune Cell Death

Published on: January 20, 2023

3.7K

Increased macrophage activation mediated by caspase recruitment domain 6 knockdown through negatively targeting AMPK

Fangyuan Chen1, Juanli Li1, Gang Tian1

  • 1Department of Cardiovascular Medicine, First Affiliated Hospital of Xi'an Jiaotong University, 710061, China.

Biochemical and Biophysical Research Communications
|February 28, 2020
PubMed

Insights

Caspase recruitment domain 6 (CARD6) protects against macrophage activation. Reduced CARD6 expression promotes inflammation and foam cell formation by affecting cholesterol metabolism and AMP-activated protein kinase (AMPK) signaling.

Area of Science:

  • Cardiovascular Biology
  • Immunology
  • Cellular Metabolism

Background:

  • Caspase recruitment domain 6 (CARD6) is linked to immunity and cancer.
  • Its role in macrophage activation and cardio-metabolic diseases is unclear.
  • Macrophage activation is critical in atherosclerosis development.

Purpose of the Study:

  • To investigate the function of CARD6 in macrophage activation.
  • To elucidate the molecular mechanisms underlying CARD6's role in macrophage polarization and foam cell formation.

Main Methods:

  • Quantitative RT-PCR and Western blot analysis to assess CARD6 expression.
  • Immunofluorescence staining to localize CARD6 in macrophages.
  • Loss-of-function studies (CARD6 knockdown) in macrophages.
  • Assessment of macrophage polarization markers (M1/M2).
  • Measurement of cholesterol uptake and efflux.
  • Analysis of AMP-activated protein kinase (AMPK) signaling pathway.

Main Results:

  • CARD6 expression decreased in macrophages from ApoE-deficient mice and those treated with oxidized LDL (OX-LDL).
  • CARD6 deficiency promoted pro-inflammatory M1 macrophage polarization and impaired M2 macrophage differentiation.
  • CARD6 knockdown enhanced cholesterol uptake, reduced cholesterol efflux, and increased foam cell formation.
  • AMPK signaling was downregulated upon CARD6 knockdown, mediating its effects on macrophage activation.

Conclusions:

  • CARD6 plays a protective role against macrophage activation and foam cell formation.
  • CARD6 exerts its effects partly through the AMPK-dependent pathway.
  • Targeting CARD6 may offer a therapeutic strategy for cardio-metabolic diseases involving macrophage dysfunction.

Keywords:
AMPKAtherosclerosisCARD6Macrophage

More Related Videos

Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages08:41

Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages

Published on: April 6, 2022

3.0K
Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry05:29

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Published on: March 24, 2023

5.3K

Related Concept Videos

Caspases01:24

Caspases

Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
13.5K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
7.7K
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
5.1K
cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
8.0K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
11.6K
The Extrinsic Apoptotic Pathway01:17

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...
7.8K