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Objective:To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) signaling pathway in propofol postconditioning-induced reduction of hippocampal neuron injury in a rat model of oxygen-glucose deprivation and restoration (OGD/R).Methods:The hippocampal neurons were isolated from fetal rats of Wistar rats at 16-18 days of gestation and primarily cultured for 7 days and then divided into 4 groups ( n=42 each) using a random number table method: control group (group C), OGD/R group (group O), propofol post-conditioning group (group P) and Nrf2 siRNA(-) transfection group (group N). The cells were routinely cultured in group C. The cells were subjected to oxygen-glucose deprivation for 1 h followed by oxygen and glucose supply in group O. Propofol (final concentration 1.2 μg/ml) was added immediately after oxygen and glucose supply, the cells were then cultured for 2 h, and the culture medium was replaced with the normal culture medium in group P. The primarily cultured neurons were transfected with Nrf2 gene knockout lentivirus on 3rd day of culture, 24 h later the cells were then routinely cultured, and the model was prepared and propofol conditioning was performed on 7th day. Cells were collected at 24 h of incubation for determination of the cell apoptosis (by flow cytometry), expression of Nrf2 and NLRP3 mRNA and protein (using quantitative real-time polymerase chain reaction or Western blot), concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β, and activities of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) (kit method). Results:Compared with group C, the apoptosis rate of neurons was significantly increased, concentrations of TNF-α, IL-6 and IL-1β were increased, the levels of GSH, SOD and CAT were decreased, the expression of Nrf2 and NLRP3 protein and mRNA was up-regulated, and the nuclear/plasma ratio of Nrf2 was increased in O and P groups ( P<0.05). Compared with group O, the apoptosis rate of neurons was significantly decreased, the concentrations of TNF-α, IL-6 and IL-1 β were decreased, the levels of GSH, SOD and CAT were increased, the expression of Nrf2 protein and mRNA was up-regulated, the nuclear/plasma ratio of Nrf2 was increased, and the expression of NLRP3 protein and mRNA was down-regulated in group P ( P<0.05). Compared with group P, the apoptosis rate of neurons was significantly increased, concentrations of TNF-α, IL-6 and IL-1β were increased, the levels of GSH, SOD and CAT were decreased, the expression of Nrf2 protein and mRNA was down-regulated, the nuclear/plasma ratio of Nrf2 was decreased, and the expression of NLRP3 protein and mRNA was up-regulated in group N ( P<0.05). Conclusions:Nrf2/NLRP3 signaling pathway is involved in propofol postconditioning-induced reduction of hippocampal neuron injury in a rat model of OGD/R.
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Objective:To compare the myocardial protection in pediatric patients undergoing living-donor liver transplantation (LDLT) performed under propofol- versus desflurane-based anesthesia. Methods:Sixty American Society of Anesthesiologists Physical Status classification Ⅲ or Ⅳ pediatric patients of both sexes, aged 5-24 months, weighing 5-15 kg, scheduled for elective LDLT under general anesthesia, were divided into 2 groups ( n=30 each) using a random number table method: propofol group (group P) and desflurane anesthesia group (group D). During anesthesia maintenance, propofol 5-10 mg·kg -1·min -1 was intravenously infused in group P, desflurane 0.65 MAC-1.30 MAC was inhaled in group D. At 5 min after induction of anesthesia, at 1 h of reperfusion, at the end of surgery, at 1, 2, 3, 7 and 14 days after surgery, and on the day of discharge, the concentrations of serum high-sensitivity cardiac troponin I, creatine kinase isoenzyme, N-terminal pro-B-type natriuretic peptide were determined by enzyme-linked immunosorbent assay, the occurrence of nausea and vomiting, agitation, and shivering, postoperative tracheal extubation time, intensive care unit stay time, and postoperative length of hospital stay were recorded within 24 h after surgery. Results:Compared with group P, the concentrations of serum high-sensitivity cardiac troponin I and creatine kinase isoenzyme were significantly decreased after surgery, the extubation time and intensive care unit stay time were shortened ( P<0.05), and no significant change was found in serum N-terminal pro-B-type natriuretic peptide concentrations, postoperative length of hospital stay and incidence of postoperative adverse effects at each time point in group D ( P>0.05). Conclusions:Desflurane has better myocardial protection than propofol in pediatric patients undergoing LDLT, which is helpful for early prognosis.
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The review summarizes the risk factors, diagnostic criteria and perioperative control strategies of acute kidney injury in pediatric liver transplantation, aiming to provide rationales for proper managements.
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Objective To evaluate the role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation (OLT).Methods Fifty-six pathogen-free healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 220-250 g,were used in the study.Thirty-two rats were selected and assigned into 4 groups (n =8 each) using a random number table:sham operation group (group S),OLT group,hydrogen-rich saline group (group HS) and chloroquine group (group CQ).The other 24 rats severed as the donors.In group S,laparotomy was performed,and the related blood vessels were isolated.The model of OLT was established in OLT,HS and CQ groups.In group OLT,normal saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group HS,hydrogen-rich saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group CQ,autophagy inhibitor chloroquine 60 mg/kg was injected intraperitoneally at 1 h before establishment of the model,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,the rats were sacrificed and hippocampi were isolated for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity,for pathological examination (with light microscope),and for detection of cell apoptosis (by TUNEL staining) and expression of autophagy-and apoptosis-related proteins caspase-3,cytochrome c (Cyt c),microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p53 in hippocampal tissues (by Western blot analysis).Apoptosis index (AI) was calculated.Results Compared with group S,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3,Cyt c,LC3 Ⅱ,Beclin-1 and p53 was up-regulated in OLT,HS and CQ groups (P＜0.05).Compared with group OLT,the MDA content and AI were significantly decreased,the SOD activity was increased,the expression of caspase-3 and Cyt c was down-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was up-regulated in group HS (P＜0.05).Compared with group HS,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3 and Cyt c was up-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was down-regulated in group CQ (P＜0.05).Conclusion The mechanism by which hydrogen inhibits apoptosis in hippocampal neurons is related to promotion of autophagy in a rat model of OLT.
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Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in attenuation of ischemia injury by hydrogen-rich University of Wisconsin (HRUW) solution during cold storage of rat donor kidneys.Methods Forty healthy adult male Wistar rats,weighing 200-250 g,were divided into 4 groups (n =10 each) using a random number table:control group (group C),University of Wisconsin (UW) solution group (group UW),HRUW solution group (group HRUW) and Nrf2 inhibitor all-trans retinoic acid (ATRA) group (group ATRA).ATRA 7 mg/kg was intraperitioneally injected once a day for 2 consecutive days,kidneys were isolated and underwent cold storage at 8 h after the last administration,and kidneys were stored in HRUW solution for 48 h at 4 ℃C in group ATRA.In UW and HRUW groups,the equal volume of normal saline was intraperitioneally injected instead,and isolated kidneys were stored in UW solution and HRUW solution for 48 h at 4 ℃C,respectively.Kidney specimens were obtained for microscopic examination and for determination of tumor necrosis factoralpha (TNF-α),interleukin-lbeta (IL-1β3),high-mobility group box 1 protein (HMGB1),IL-10 and 8-iso-prostaglandin F2α (8-iso-PGF2α) contents (by enzyme-linked immunosorlbent assay),superoxide dismutase (SOD) and catalase (CAT) activities (using spectrophotometry),and expression of Nrf2,heme oxygenase-1 (HO-1),Bcl-2,Bax and caspase-3 in renal tissues (by using Western blot).The damage to the renal tubules was scored.Results Compared with group C,renal tubular damage scores were signifieantly increased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were increased,IL-10 contents were decreased,the expression of Nrf2 and HO-1 was up-regulated,SOD and CAT activities were decreased,the expression of Bcl-2 was down-regulated,and the expression of Bax and caspase-3 was upregulated in group UW (P＜0.05 or 0.01).Compared w,ith group UW,renal tubular damage scores were significantly decreased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were decreased,IL-10 contents were increased,the expression of Nrf2 and HO-1 was up-regulated,SOD and CAT activities were increased,the expression of Bcl-2 was up-regulated,and the expression of Bax and caspase-3 was down-regulated in group HRUW,and the expression of Nrf2 and Bcl-2 was up-regulated (P＜0.05),and no significant change was found in the other parameters in group ATRA (P＞0.05).Compared witb group HRUW,renal tubular damage seores were significantly increased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were increased,IL-10 contents were decreased,the expression of HO-1 and Bcl-2 was down-regulated,SOD and CAT activities were decreased,and the expression of Bax and caspase-3 was up-regulated in group ATRA.Conclusion HRUW solution reduces inflammatory responses,oxidative damage and cell apoptosis during cold storage of rat donor kidneys,and the mechanism by which HRUW solution attenuates ischemia injury is related to activation of Nrf2 signaling pathway.
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Objective To investigate the effect of berberine pretreatment on hypoxia/reoxygenation (H/R)-induced apoptosis in human renal tubular epithelial cells.Methods The human renal tubular epithelial cells were cultured and seeded in culture dishes (2 ml/dish) or 96-well plates (200 μl/well) with the density of 1 ×106/ml.The cells were then randomly divided into 4 groups (n =30 each):normal control group (group C),berberine group (group B),H/R group and H/R + berberine group (H/R + B group).In groups B and H/R + B,berberine 10 μmol/L was added to the culture medium and the cells were incubated for 2 h.Groups H/R and H/R + B were then exposed to 94％ N2-5％ CO2-1％ O2 for 24 h followed by 3 h reoxygenation.The cell viability,apoptotic rate and level of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected.The expression of caspase-3,activated caspase-3,cytochrome c,glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was determined.The ratio of Bax/Bcl-2 was calculated.Results Compared with group C,the cell viability,SOD activity and caspase-3 expression were significantly decreased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were increased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was up-regulated in groups H/R and H/R + B (P ＜ 0.05).Compared with group H/R,the cell viability,SOD activity and caspase-3 expression were significantly increased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were decreased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was down-regulated in group H/R + B (P ＜ 0.05).Conclusion Berberine pretreatment can inhibit H/R-induced apoptosis in human renal tubular epithelial cells,and inhibition of mitochondrial stress pathway and endoplasmic reticulum stress pathway is involved in the mechanism.
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Objective To investigate the effects of hydrogen-rich HC-A solution,the self-made kidney preservation solution,on renal cold ischemia/reperfusion (I/R) injury in rats.Methods Twenty-four healthy male Wistar rats,aged 8-10 weeks,weighing 200-250 g,were randomly divided into 3 groups (n =8 each):control group (H1 group),common kidney preservation solution group (H2 group) and hydrogen-rich HC-A kidney preservation solution group (H3 group).In H1 group,only the right kidney was removed.In H2 group,the left kidney was perfused with and cold stored in 4 ℃ common HC-A kidney preservation solution.In H3 group,the left kidney was perfused with and cold stored in a container filled with 4 ℃ common HC-A kidney preservation solution.Blood samples were obtained from the inferior vena cava at 24 h of reperfusion to detect the levels of serum blood urea nitrogen (BUN),creatinine (Cr),TNF-α and IL-6.Left kidneys were removed for determination of malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) contents and for examination of the pathological changes in renal tissues (by light microscopy).Results The levels of serum BUN,Cr,TNF-α and IL-6 and contents of MDA and 8-OHdG were significantly higher in H2 and H3 groups than in H1 group,and lower in H3 group than in H2 group (P ＜ 0.05).There were no significant pathological changes in renal tissues in H1 group,the damage to renal tubules was obvious in H2 group and the damage to renal tubules was significantly ameliorated in H3 group as compared with H2 group.Conclusion Hydrogen-rich kidney preservation solution can attenuate renal cold I/R injury in rats.