Alda-1 alleviates brain injury after cardiopulmonary resuscitation by regulating acyl-CoA synthetase long-chain family member 4/glutathione peroxidase 4 pathway-mediated ferroptosis in swine / 中华危重病急救医学

OBJECTIVE@#To investigate whether the acetaldehyde dehydrogenase 2 specific activator, Alda-1, can alleviate brain injury after cardiopulmonary resuscitation (CPR) by inhibiting cell ferroptosis mediated by acyl-CoA synthetase long-chain family member 4/glutathione peroxidase 4 (ACSL4/GPx4) pathway in swine.@*METHODS@#Twenty-two conventional healthy male white swine were divided into Sham group (n = 6), CPR model group (n = 8), and Alda-1 intervention group (CPR+Alda-1 group, n = 8) using a random number table. The swine model of CPR was reproduced by 8 minutes of cardiac arrest induced by ventricular fibrillation through electrical stimulation in the right ventricle followed by 8 minutes of CPR. The Sham group only experienced general preparation. A dose of 0.88 mg/kg of Alda-1 was intravenously injected at 5 minutes after resuscitation in the CPR+Alda-1 group. The same volume of saline was infused in the Sham and CPR model groups. Blood samples were collected from the femoral vein before modeling and 1, 2, 4, 24 hours after resuscitation, and the serum levels of neuron specific enolase (NSE) and S100 β protein were determined by enzyme-linked immunosorbent assay (ELISA). At 24 hours after resuscitation, the status of neurologic function was evaluated by neurological deficit score (NDS). Thereafter, the animals were sacrificed, and brain cortex was harvested to measure iron deposition by Prussian blue staining, malondialdehyde (MDA) and glutathione (GSH) contents by colorimetry, and ACSL4 and GPx4 protein expressions by Western blotting.@*RESULTS@#Compared with the Sham group, the serum levels of NSE and S100β after resuscitation were gradually increased over time, and the NDS score was significantly increased, brain cortical iron deposition and MDA content were significantly increased, GSH content and GPx4 protein expression in brain cortical were significantly decreased, and ACSL4 protein expression was significantly increased at 24 hours after resuscitation in the CPR model and CPR+Alda-1 groups, which indicated that cell ferroptosis occurred in the brain cortex, and the ACSL4/GPx4 pathway participated in this process of cell ferroptosis. Compared with the CPR model group, the serum levels of NSE and S100 β starting 2 hours after resuscitation were significantly decreased in the CPR+Alda-1 group [NSE (μg/L): 24.1±2.4 vs. 28.2±2.1, S100 β (ng/L): 2 279±169 vs. 2 620±241, both P < 0.05]; at 24 hours after resuscitation, the NDS score and brain cortical iron deposition and MDA content were significantly decreased [NDS score: 120±44 vs. 207±68, iron deposition: (2.61±0.36)% vs. (6.31±1.66)%, MDA (μmol/g): 2.93±0.30 vs. 3.68±0.29, all P < 0.05], brain cortical GSH content and GPx4 expression in brain cortical was significantly increased [GSH (mg/g): 4.59±0.63 vs. 3.51±0.56, GPx4 protein (GPx4/GAPDH): 0.54±0.14 vs. 0.21±0.08, both P < 0.05], and ACSL4 protein expression was significantly decreased (ACSL4/GAPDH: 0.46±0.08 vs. 0.85±0.13, P < 0.05), which indicated that Alda-1 might alleviate brain cortical cell ferroptosis through regulating ACSL4/GPx4 pathway.@*CONCLUSIONS@#Alda-1 can reduce brain injury after CPR in swine, which may be related to the inhibition of ACSL4/GPx4 pathway mediated ferroptosis.

Effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine / 中华麻醉学杂志

Objective:To evaluate the effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine.Methods:Twenty-two healthy male white swine, weighing 35-43 kg, were divided into 3 groups using a random number table method: sham operation group (group S, n=6), cardiac arrest-cardiopulmonary resuscitation group (group CA-CPR, n=8) and Alda-1 group ( n=8). The animals only underwent the general preparation in group S, and the swine model of cardiac arrest and cardiopulmonary resuscitation was developed by 8 min of electrically induced cardiac arrest through the pacing catheter in the right ventricle followed by 8 min of cardiopulmonary resuscitation in CA-CPR and Alda-1 groups.Alda-1 0.88 mg/kg was intravenously injected at 5 min after resuscitation in group Alda-1, and the equal volume of vehicle was administered instead in the other two groups.Stroke volume (SV) and global ejection fraction (GEF) were measured using PiCCO before developing the model and at 1, 2 and 4 h after resuscitation (T 0-3). Venous blood samples were collected from the femoral vein to measure the concentrations of serum cardiac troponin (cTnI) by enzyme-linked immunosorbent assay at T 0-3, and at 24 h after resuscitation (T 4). The animals were then sacrificed, and myocardial tissues in the left ventricle were harvested to measure the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) (by Western blot), iron deposition (by Prussian blue staining), 4-hydroxy-2-nonenal (4-HNE) content (by enzyme-linked immunosorbent assay), and malondialdehyde (MDA) and glutathione (GSH) contents (by colorimetry). Results:Compared with group S, SV and GEF were significantly decreased at T 1-3, the serum concentrations of cTnI were increased at T 1-4, myocardial ACSL4 expression was up-regulated, GPX4 expression was down-regulated, iron deposition and contents of 4-HNE and MDA were increased, and the content of GSH was decreased in CA-CPR and Alda-1 groups ( P<0.05). Compared with group CA-CPR, SV and GEF were significantly increased at T 2-3, the serum concentrations of cTnI were decreased at T 3-4, myocardial ACSL4 expression was down-regulated, GPX4 expression was up-regulated, iron deposition and contents of 4-HNE and MDA were decreased, and the content of GSH was increased in group Alda-1 ( P<0.05). Conclusions:Alda-1 can alleviate myocardial injury after cardiac arrest and cardiopulmonary resuscitation in swine and further improve cardiac dysfunction, and the mechanism may be related to inhibition of cell ferroptosis.

A preliminary study on traditional Chinese medicine syndrome rating scale for acute gastrointestinal injury in sepsis / 中国中西医结合急救杂志

Objective To develop a traditional Chinese medicine (TCM) syndrome score scale for acute gastrointestinal injury (AGI) in sepsis, and to carry out its reliability and validity analyses and its clinical preliminary application. Methods ① According to the characteristics of intensive care unit (ICU) patients, combined with the understanding of etiology, pathogenesis and physical signs of TCM and literature search, a preliminary framework of scoring system for TCM syndromes of AGI in sepsis was constructed to carry out the scoring by this scale. ② After the scale and data were obtained, the analyses of split-half reliability (indicated by Guttman's split-half reliability of the a and b groups), test-retest reliability and the internal consistency reliability (expressed by the Cronbach's coefficient α) were carried out, and the structural validity and criterion validity were also analyzed. ③ The AGI patients were divided into two groups according to the 28-day survival and death conditions, and the AGI TCM syndrome score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, and multiple organ dysfunction syndrome (MODS) score were compared between the two groups to determine the best cut-off point for survival analysis. Results ① The first draft of the septic AGI TCM syndrome rating scale was prepared, The TCM syndrome indicators include: abdominal distension, constipation/diarrhea, diet situation, vomiting/stomach retention, tongue proper, tongue coating, pulse manifestation, belching, body temperature, and accompanied syndrome, there were 6 points for scoring, 0 - 6 points, and they were divided into normal (0 points), mild (2 points), moderate (4 points), and severe (6 points) in severity. ② Eighty-eight patients with septic AGI were included in the final statistics. The retest of correlation coefficient of this scale was R = 0.974 (> 0.85), Guttman's split-half reliability was 0.793 (> 0.7) and the Cronbach's coefficient α was > 0.7. This scale was suitable for factor analysis. After rotation, 3 factors were determined, which were named as TCM syndrome differentiation, related physical signs, and gastrointestinal tolerance. After modeling, the confirmatory factor analysis showed that the model approximate error root mean square (RMSEA) was 0.07 (< 0.08), and the goodness of fit index (CFI) = 0.90; the Pearson correlation analyses between the criteria validity of APACHE Ⅱ, SOFA, MODS scores and TCM 1 score and TCM 2 score of this scale showed that the r values were 0.802 and 0.752, 0.524 and 0.519, 0.619 and 0.590, respectively, all P < 0.01. ③ Compared with the survival group, TCM score (33.73±5.95 vs. 37.28±5.26, t = 2.945, P = 0.004), the APACHE Ⅱ score (19.90±4.47 vs. 22.28±5.79, t = 2.069, P = 0.043), SOFA score (8.73±1.11 vs. 9.64±1.38, t = 3.329, P = 0.020) in the death group were significantly decreased; MODS score in the death group showed a decreasing trend (6.65±1.22 vs. 7.28±1.60, t = 2.078, P = 0.050). Cox regression analysis showed that when the survival analysis was performed with a cut-off point of 35, the 28-day survival rate of patients with TCM syndrome score ≥ 35 was significantly lower than that of patients with < 35 score, χ2= 6.362, P = 0.012. Conclusions The TCM syndrome rating scale for AGI in sepsis was successfully prepared. The statistical reliability and validity of this scale are good. Preliminary clinical application shows that this scale can predict the prognosis and severity of patients with septic AGI. Trial registration China Clinical Trial Registry Center, ChiCTR-IOR-15007625.