AICAR suppresses TNF-α-induced complement factor B in RPE cells Scientific Reports

Fresh pancreatic and liver tissues and blood samples were collected for biochemical analysis. The levels of serum amylase and lipase were measured by assay kit (C , A , Nanjing Jiancheng Bioengineering Institute, Nanjing, China) to evaluate the degree of pancreatitis. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were https://www.teatappetimoderni.it/steroids-understanding-their-use-benefits-and-79/ measured with commercial kit (C , C , Nanjing Jiancheng Bioengineering Institute, Nanjing, China) to evaluate the degree of liver injury and function.

• Based on the causal role of JNK activation in AICAR-triggered apoptosis, inhibition of JNK was likely a key contributor in the non-apoptotic effect of metformin.
• Fourteen-week-old male, lean (L; 31.3 g body wt) wild-type andob/ob (O; 59.6 g body wt) mice are injected with the AMP-activated kinase (AMPK) activator AICAR (A) at 0.5 mg/g per day or saline control (C) for 14 days.
• Continuous in vitro culture of MCSs ultimately causes undesirable outcomes and reduced efficacy of MSC-based therapies 3,4,5.
• AICAR or 5-Aminoimidazole-4-carboxamide ribonucleotide, is a synthetic adenosine monophosphate analog.
• It highlights NAD+’s involvement in cellular energy production, DNA repair, and sirtuin activation, which contribute to improved metabolic efficiency, energy levels, and cognitive function.

The central goal of this study was to investigate the pharmacological activation of AMPK by AICAR as a therapeutic strategy for the treatment of PALI. Our findings demonstrated that AICAR activates AMPK, which leads to Nrf2-mediated antioxidant stress and inhibition of NLRP3-related inflammation, and thus improving PALI. This study indicated that AMPK exerted an essential role in the pathological processes of PALI and presented the first evidence that pharmacological activation of AMPK by AICAR ameliorates PALI, suggesting that AICAR may be a promising therapeutic agent for the treatment of PALI.

Population doubling time assay

However, we observed that a combination of everolimus and AICAR induced only a slight, non-significant, increase in clonogenic killing activity compared to either agent alone. Moreover, the simultaneous administration of AICAR and everolimus did not enhance the radiosensitization caused by either drug alone. AMPK activation by AICAR suppresses the mTOR signalling pathway 47, leading to decreased proliferation and survival. Thus, it is possible that administration of AICAR alone is sufficient to abrogate the mTOR hyperactivity in PC3 cells caused by inactive PTEN, making addition of an mTOR inhibitor redundant. AMPK is the evolutionarily conserved master energy sensor/regulator of the eukaryotic cells 23, 48.

AICAR and metformin inhibited IκB phosphorylation following stimulation by TNF-α in KGN cells

We thus continued to use this model to dissect the effects of AICAR/Compound C on AMPK in T cells. We first measured the AMPK activation using resting T cells from lymph nodes of WT and KO mice. Intracellular staining of phosphorylation of AMPK Thr-172 (p-AMPK) showed that AMPK was not or only weakly activated in resting WT T cells as compared to KO T cells. Interestingly, treatment with AICAR significantly increased phosphorylation of AMPK in WT T cells, but not in KO T cells, suggesting a specific activation of AMPK with AICAR. We did not observe any obvious inhibition of p-AMPK with Compound C treatment (Figure 1A), which may be due to the non- or weak activation of AMPK in resting T cells.

Followed by incubation in ECL Western Blotting Substrate (Pierce) for 5 min and visualised on ChemiDoc Imaging System (BioRad). Cells were harvested from cultured dishes and were lysed in a lysis buffer 20 mM Tris-HCl pH 7.6, 1 mM EDTA, 140 mM NaCl, 1% NP-40, 1% aprotinin, 1 mM phenylemethylsulfonyl fluoride (PMSF), 1 mM sodium vanadate. Cell lysates (40 µg protein/line) were separated on a 5 to 20% Tris-Tricine Ready Gel SDS-PAGE (Bio-Rad) for nitrocellulose membrane blotting.

There is a body of evidence showing that aged cells have reduced autophagy and AMPK activity. It has also been demonstrated that suppressed autophagy is the main factor in the pathogenesis of aging in several tissues and cell types, especially in stem cells 18, 48, 49, 57, 58. For instance, García-Prat et al. showed that treating old mice with a rapamycin regimen increased autophagy and re-established cell proliferation 18.

Sucrose density gradient centrifugation was employed to analyze muscle polysome aggregation state following treadmill exercise. Homogenates were incubated on ice for 5 min, 150 μl/ml Tween-deoxycholate mix (1.34 ml Tween 20, 0.66 g deoxycholate, and 18 ml sterile water) was added, and the samples were thoroughly mixed. Samples were incubated on ice for 15 min and then centrifuged at 1,000 g for 15 min at 4°C. The resulting supernatant (600 μl) was layered on a 20%-47% linear sucrose density gradient 50 mM HEPES (pH 7.4), 75 mM KCl, and 5 mM MgCl2 and centrifuged in a SW41 rotor at 40,000 rpm for 4 h at 4°C. After centrifugation, the gradient was displaced upward (2 ml/min) using Fluorinert (Isco, Lincoln, NE) through a spectrophotometer, and the optical density at 254 nm was continuously recorded (150 cm/h chart speed).

Further, forced-activation of AMPK, by adding two other AMPK activators (A and Compound 13), or expressing a constitutively-active mutant AMPKalpha (T172D), didn’t induce GBC-SD cell death. Remarkably, AICAR treatment in gallbladder cancer cells induced endoplasmic reticulum (ER) stress activation, the latter was tested by caspase-12 activation, C/EBP homologous protein (CHOP) expression and IRE1/PERK phosphorylation. Contrarily, salubrinal (the ER stress inhibitor), z-ATAD-fmk (the caspase-12 inhibitor) or CHOP shRNAs significantly attenuated AICAR-induced gallbladder cancer cell apoptosis. Together, we conclude that AICAR-induced gallbladder cancer cell apoptosis requires ER stress activation, but is independent of AMPK. Even though peptide- and siRNA-based approaches have the potential to target MUC1, small molecule therapeutics have many advantages due to their cell-permeable and potent features in the clinical treatment 24, 100. Our study was the first to describe the intrinsic metabolite AICAR physically binding and targeting MUC1 to induce lung tumour cell apoptosis.