Cell lines

Microglia cells

Preparation Method

Microglia cells (1×105) were stimulated with the caspase-1 inhibitor Ac-YVAD-CMK (dissolved by DMSO) for 1h. Thrombin was added at a final concentration of 10U/mL to activate the microglia cells for 24h.

Reaction Conditions

40μmol/L or 80μmol/L

Applications

High level Ac-YVAD-CMK treatment (40μmol/L or 80μmol/L) significantly decreased the mRNA levels of IL-1β/IL-18. The 40μmol/L dose of Ac-YVAD-CMK significantly reduced the protein levels of caspase-1 (p20) and mature IL-1β/IL-18 compared with the thrombin-activated microglia group.

Animal models

Male Sprague Dawley rats，weighing 250-275g

Preparation Method

Ac-YVAD-CMK (300ng/rat in 3mL) was injected intracerebroventricularly 10 min after permanent middle cerebral artery occlusion in the rat.

Dosage form

300ng/rat in 3mL, intracerebroventricular injection

Applications

Ac-YVAD-CMK treatment induced a significant reduction of infarct volume not only 24 h after ischemia but also 6d later. Ac-YVAD-CMK treatment resulted in a reduction not only of caspase-1 but also of caspase-3 activity at 24h and led to a parallel decrease of apoptosis as measured by nucleosome quantitation. Likewise, brain levels of the proinflammatory cytokines IL-1b and TNF-a were reduced at 24h.

• J Clin Med 11.20 (2022): 6204. PMID:36294525

Ac-YVAD-CMK is a selective irreversible inhibitor of caspase-1 (Ki=0.8nM), which can prevent the proinflammatory cytokine IL-1β activation. Ac-YVAD-CMK can reduce the inflammatory response and induce a long-lasting neuroprotective effect^[1,2,3].

Ac-YVAD-CMK reduced the expression of IL-1β and IL-18 in activated microglia in vitro^[3]. Ac-YVAD-CMK(40μM) decreased the rapid cell (PAMs, 3D4/21 cells and the endothelial cell line) death induced by ApxⅠ^[4]. Ac-YVAD-CMK attenuated the inhibitory effects of berberine on the viability, migration and invasion of HepG2 cells^[5].

AC-YVAD-CMK treatment significantly alleviated sepsis-induced acute kidney injury, with decreased histological injury in renal tissues, suppressed the accumulation of neutrophils and macrophages in renal tissues, and decreased sCR and BUN level^[6]. Ac-YVAD-CMK protected the brain against ICH-induced injury, and the neuroprotective effect may result from anti-inflammation-induced blood–brain barrier protection^[7]. Pretreatment of rats with Ac-YVAD-CMK (12.5mM/kg) significantly reduced endotoxin-induced mortality from 83% to 33% using Log Rank analysis^[8].

References:
[1] Wang F, Li G, et al. Alcohol accumulation promotes esophagitis via pyroptosis activation. Int J Biol Sci. 2018;14(10):1245-1255. Published 2018 Jul 13.
[2] Rabuffetti M, Sciorati C, et al. Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines. J Neurosci. 2000;20(12):4398-4404.
[3] Liang H, Sun Y, et al. Ac-YVAD-cmk improves neurological function by inhibiting caspase-1-mediated inflammatory response in the intracerebral hemorrhage of rats. Int Immunopharmacol. 2019;75:105771.
[4] Hernandez-Cuellar E, Guerrero-Barrera AL, et al. An in vitro study of ApxI from Actinobacillus pleuropneumoniae serotype 10 and induction of NLRP3 inflammasome-dependent cell death. Vet Rec Open. 2021;8(1):e20. Published 2021 Oct 4.
[5] Chu Q, Jiang Y, et al. Pyroptosis is involved in the pathogenesis of human hepatocellular carcinoma. Oncotarget. 2016;7(51):84658-84665.
[6] Yang M, Fang JT, et al. Caspase-1-Inhibitor AC-YVAD-CMK Inhibits Pyroptosis and Ameliorates Acute Kidney Injury in a Model of Sepsis. Biomed Res Int. 2021;2021:6636621.
[7] Wu B, Ma Q, et al. Ac-YVAD-CMK Decreases Blood-Brain Barrier Degradation by Inhibiting Caspase-1 Activation of Interleukin-1β in Intracerebral Hemorrhage Mouse Model. Transl Stroke Res. 2010;1(1):57-64.
[8] Mathiak G, Grass G, et al. Caspase-1-inhibitor ac-YVAD-cmk reduces LPS-lethality in rats without affecting haematology or cytokine responses. Br J Pharmacol. 2000;131(3):383-386.