vx-765 and Reperfusion-Injury

Intestinal ischemia-reperfusion injury (IIRI) is a common clinical event that can cause serious consequences. The study aimed to investigated the effect of VX-765 in IIRI and its mechanism.. The hypoxia-reoxygenation (H/R) cell model and IIRI mouse model were generated to examine the in vitro and in vivo effects of VX-765 on IIRI. IIRI was evaluated by histological assessment. ELISA was performed to determine the levels of IL-6, TNF-α, IL-1β, caspase-1, and GSDMD in intestinal tissues as well as the levels of MDA, SOD, CAT, caspase-1, and GSDMD in Caco-2 cells. Relative protein levels of NLRP3, ASC, IL-18, IL-1β, cleaved Caspase1, and GSDMD-N were analyzed by Western blotting. CCK-8 Assay was conducted to determine the optimal concentration of VX-765 for the in vitro studies. Flow cytometry, fluorescence microscopy and real-time PCR (RT-PCR) were used to assess ROS levels and the mRNA levels of IL-18 and IL-1β, respectively. Immunofluorescence staining was performed to examine the subcellular localization of P65 and NLRP3.. VX-765 reduced IIRI-induced oxidative stress and inflammatory response both in vivo and in vitro, while it decreased the levels of TNF-α, IL-6, IL-1β as well as the modified Park/Chiu scores. The optimal concentration of VX-765 for the in vitro studies was 10 μM. Moreover, VX-765 inhibited the nuclear translocation of P65, reduced oxidative stress and down-regulated the activation of NLRP3 inflammasome.. VX-765 prevents IIRI presumably by inhibiting the activation of NLRP3 inflammasome.

Topics: Animals; Caco-2 Cells; Dipeptides; Humans; Inflammasomes; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; para-Aminobenzoates; Reperfusion Injury

Topics: Animals; Caspase 1; Caspase Inhibitors; Cells, Cultured; Dipeptides; Endothelial Cells; Inflammasomes; Inflammation; Lung; Lung Injury; Male; Mice; Mice, Inbred C57BL; para-Aminobenzoates; Pyroptosis; Reperfusion Injury; Signal Transduction

Pyroptosis is a newly identified lytic form of programmed cell death which is characterized by plasma membrane blebbing and rupture. Pyroptosis occurs in cerebral ischemia injury, and contributes to the activation and secretion of the inflammatory cytokines interleukin (IL)-1β, IL-18, and IL-6. Previous reports have found that Dendrobium alkaloids (DNLA) can exert neuroprotective effects against oxygen-glucose deprivation/reperfusion (OGD/R) damage in vitro, but the mechanisms underlying these effects remain elusive. In this study, we investigated whether DNLA exerted therapeutic benefits against cerebral ischemia-reperfusion (CIR) damage via ameliorating pyroptosis and inflammation. OGD/R damage was induced in HT22 cells pretreated with DNLA (0.03, 0.3, or 3 mg/ml, 24 h prior to OGD/R), MCC950 (10 ng/ml, 1 h prior), and VX765 (10 ng/ml, 1 h prior). Neuronal apoptosis, necrosis, pyroptosis, and pathological changes were analyzed 24 h following OGD/R. Further to this, male C57/BL mice pretreated with different concentrations of DNLA (0.5 or 5 mg/kg, ip.) for 24 h and VX765 (50 mg/kg, ip., 1 h before CIR) underwent transient middle cerebral artery occlusion and reperfusion. We found that DNLA pretreatment resulted in a lower neurologic deficit score, a reduced infarct volume, fewer pyroptotic cells, and reduced levels of inflammatory factors 24 h after CIR. Furthermore, DNLA administration also reduced the levels of the pyroptosis-associated proteins Caspase-1 and gasdermin-D, particularly in the hippocampal CA1 region. Similar decreases were observed in the levels of the inflammatory factors IL-1β, IL-6, and IL-18. OGD/R-associated ultrastructural damage was seen to improve following DNLA administration, likely due to the regulation of the tight junction protein Pannexin-1 by DNLA. Overall, these findings demonstrate that DNLA can protect against CIR damage through inhibiting pyroptosis-induced neuronal death, providing new therapeutic insights for CIR injury.

Topics: Alkaloids; Animals; Caspase 1; Caspase Inhibitors; Cell Line; Dipeptides; Hippocampus; Infarction, Middle Cerebral Artery; Intracellular Signaling Peptides and Proteins; Male; Mice, Inbred C57BL; Necrosis; Neuroprotective Agents; para-Aminobenzoates; Phosphate-Binding Proteins; Pyroptosis; Reperfusion Injury