1.Effect of edaravone on apoptosis in hippocampal cells in a rat model of endotoxic shock
Chinese Journal of Anesthesiology 2015;35(7):862-865
Objective To evaluate the effect of edaravone on apoptosis in hippocampal cells in a rat model of endotoxic shock.Methods Thirty-six male Sprague-Dawley rats, weighing 200-250 g, aged 6 weeks, were randomly divided into 3 groups (n=12 each) using a random number table: control group (group C), endotoxic shock group (group ES), and edaravone group (group E).Lipopolysaccharide 10 mg/kg was injected via the femoral vein to establish the model of endotoxic shock in ES and E groups, while the equal volume of normal saline was given in group C.In group E, edaravone 3 mg/kg was intravenously injected immediately after establishment of the model once every 2 h until the animals were sacrificed.The equal volume of normal saline was given instead of edaravone in C and ES groups.At 6 and 12 h after administration of edaravone, 6 rats in each group were sacrificed, and the hippocampi were isolated for determination of malondialdehyde (MDA) content (using thiobarbituric acid method) , tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) contents (using enzyme-linked immunosorbent assay), and cell apoptosis in hippocampal CA1 region (by TUNEL assay).The apoptotic index was calculated.Results Compared with group C, the MDA, TNF-α and IL-6 contents were significantly increased at 6 and 12 h after administration of edaravone, and the apoptotic index was increased at 12 h after administration of edaravone in ES and E groups.Compared with group ES, the MDA, TNF-α and IL-6 contents were significantly decreased at 6 and 12 h after administration of edaravone, and the apoptotic index was decreased at 12 h after administration of edaravone in group E.Conclusion Edaravone can reduce apoptosis in hippocampal cells, and the mechanism is associated with the reduced oxidative stress and inflammatory responses in a rat model of endotoxic shock.
2.Effect of edaravone on permeability of blood-brain barrier in septic rats
Chinese Journal of Anesthesiology 2013;33(8):986-988
Objective To evaluate the effects of edaravone on the permeability of blood-brain barrier in septic rats.Methods Ninety male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=30 each):control group (group C),sepsis group (group lipopolysaccharide (LPS)) and edaravone group (group E).Sepsis was induced by injection of LPS 10 mg/kg via the femoral vein in LPS and E groups.After LPS injection,edaravone 3.0 mg/kg was injected intravenously every 2h for 7 times in group E.The equal volume of normal saline was administered instead of edaravone in C and LPS groups.At 2,6 and 12h after LPS injection,5 rats were chosen and Evan's blue (EB) was injected via the femoral vein,and then the rats were sacrificed and brain tissues were removed for determination of EB and water contents.Another 5 rats were chosen and blood samples were taken from the femoral artery for measurement of serum MDA concentration,and then the rats were sacrificed and the brain tissue was harvested for microscopic examination.Results Compared with group C,brain water and EB contents were significantly increased at 6 and 12h after LPS injection,and the serum MDA concentration was increased at 2,6 and 12h after LPS injection in LPS and E groups (P < 0.05).Compared with group LPS,brain water and EB contents were significantly decreased at 6 and 12h after LPS injection,and serum MDA concentrations were decreased at 2,6 and 12h after LPS injection in group E (P < 0.05).Sepsis-induced pathological changes were significantly attenuated in group E.Conclusion Edaravone can decrease the permeability of blood-brain barrier,attenuate brain edema and brain injury in septic rats,and reduction of oxygen free radical production may be involved in the mechanism.
3.Study on the mutagenesis effects of low-dose sodium arsenite by Ames test
Chinese Journal of Endemiology 2008;27(4):389-392
Objective To test whether sodium arsenite can induce in vitro reverse mutation of Salmonella typhimurium histamine-auxotroph mutant. Methods Ames test was carded out with Salmonella typhimurium strains TA97,TA98,TA100 and TA102 by standard method with or without the liver microsomal enzyme activation system (+S9,-S9). Results At concentrations of sodium arsenite from 500.00 to 5000.00 μg/plate, no colonies were seen on the plates of TA97,TA98,TA100 or TA102, with or without the presence of S9. At concentrations of sodium arsenite of 0.01,0.10,10.00 μg/plate and with the presence of S9, twice as many colonies grew on the plates of TA102 as the negative control(P<0.05). Without S9 activation,twice as many colonies grew on the plates of TA100 as the negative control(P<0.05)at concentrations of sodium arsenite of 1.00,10.00 μg/plate(P<0.05). The reverse mutation colonies induced by sodium arsenite in TA98 strain were twice as many as negative control group at concentrations of 0.01,0.10 μg/plate(P<0.05). There was no obvious increase of the strain clones in the other(P0.05). Conclusions With and without S9 activation, the doses of 500.00,5000.00 μg/plate sodium arsenite resulted in a toxic effect and a reduction of the revertants among the strain. At concentrations of 0.01~10.00 μg/plate, sodium arsenite exhibited mutngenesis effects.
4.Effects of volume resuscitation with different fluids on expression of aquaporin 1 in pulmonary capillary endothelial cells of endotoxemic rats
Shuying WANG ; Na CHEN ; Ling PEI
Chinese Journal of Anesthesiology 2014;34(9):1108-1111
Objective To evaluate the effects of volume resuscitation with different fluids on expression of aquaporin 1 (AQP-1) in pulmonary capillary endothelial cells of endotoxemic rats.Methods Fifty pathogen-free male Sprague-Dawley,rats,weighing 250-300 g,aged 6-8 weeks,were randomly divided into 4 groups (n =10 each):control group (group C),lipopolysaccharide group (group LPS),NaCl group (group NS),6% hydroxyethyl starch 130/0.4 group (group HES) and hypertonic hydroxyethyl starch 40 group (group HSH).Sepsis was induced with LPS 5 mg/kg injected via the femoral vein.At l0 min after the model was successfully established,the corresponding fluids were infused into the femoral vein at a constant speed (15 ml/kg over 6 h) via a pump.At 0,3 and 6 h after LPS administration,blood samples were collected from the femoral artery for measurement of serum tumor necrosis factor-α (TNF-α) concentrations and for blood gas analysis.PaO2 and lactate (Lac) concentrations were recorded and oxygenation index (PaO2/FiO2) was calculated.The rats were sacrificed at 6 h after LPS administration,and lungs were removed for determination of AQP-1 expression in pulmonary capillary endothelial cells and wet-to-dry lung weight ratio (W/D ratio),and for microscopic examination of the pathological changes which were scored.Results Compared with group C,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly increased,AQP-1 expression was down-regulated,and PaO2 and oxygenation index were decreased in LPS,NS,HES and HSH groups.Compared with group LPS,W/D ratio,serum TNF-a and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in NS,HES and HSH groups.Compared with group NS,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in HES and HSH groups.Compared with group HES,the blood Lac concentrations were significantly decreased,and no significant changes were found in the other parameters in group HSH.Conclusion Volume resuscitation with NaC1,hydroxyethyl starch 130/0.4 and hypertonic hydroxyethyl starch 40 can reduce the lung injury effectively in endotoxemic rats,and hypertonic hydroxyethyl starch 40 provides more significant efficacy,and up-regulated AQP-1 expression in pulmonary capillary endothelial cells is involved in the mechanism.
5.Lentiviral vector-mediated transfection of bone morphogenetic protein 2 gene into endothelial progenitor cells from rat bone marrow
Xiuru YIN ; Ling PEI ; Zuodi LIANG
Chinese Journal of Tissue Engineering Research 2014;(32):5197-5202
BACKGROUND:Gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. The key point is to choose the proper cell, gene and vector. OBJECTIVE:To investigate the effectiveness and feasibility of bone morphogenetic protein 2 (BMP2) gene transfected into endothelial progenitor cells (EPCs) from rat bone marrow for gene therapy. METHODS:The EPCs were isolated, cultured and identified from the bone marrow of Sprague-Dawley rats. Empty vector (LV-eGFP) or BMP2 gene (LV-eGFP-BMP2) was transferred into EPCs by the constructed lentiviral vector (LV). We examined the transfection efficiency by eGFP fluorescence, BMP2 secretion by ELISA, BMP2 expression by Western blot, and compared the capacities of migration, proliferation and anti-apoptosis after transfection in the three groups of normal EPCs, empty vector-EPCs, and BMP2-EPCs. RESULTS AND CONCLUSION:The transfection efficiency of lentiviral vector was 90%. BMP2 gene-transferred EPCs secreted and expressed more BMP2 proteins (P<0.01), and showed enhanced anti-apoptotic ability (P<0.05). The proliferation and migration capacity did not change obviously (P>0.05). After successful transfection with lentivirus-BMP2 gene, EPCs can secrete and express more BMP2 protein and show enhanced anti-apoptotic ability without obvious influence on the proliferation and migration capacity.
6.Effects of preoperative sleep disturbance on efficacy of flurbiprofen for postoperative analgesia in patients undergoing endoscopic nasal surgery
Chinese Journal of Anesthesiology 2014;34(z1):71-73
Objective To investigate the effects of preoperative sleep disturbance on the efficacy of flurbiprofen for postoperative analgesia in patients undergoing endoscopic nasal surgery.Methods Ninety-six ASA Ⅰ or Ⅱ patients of both sexes (aged 20-60 years and weighing 50-80 kg) undergoing endoscopic nasal surgery were enrolled in this study.Pittsburg sleep quality index was used to evaluate the long-term sleep quality before hospitalization and Athens sleep quality index was used to evaluate the short-term sleep quality in hospital.The patients were divided into four groups according to the types of preoperative sleep disturbance (n =24 each):no sleep disturbance (group Ⅰ),long-term sleep disturbance (group Ⅱ),acute short-term sleep disturbance (group Ⅲ),and long-term + acute short-term sleep disturbance (group Ⅳ).Anesthesia was induced with sufentanil,propofol and cis-atracurium and maintained with intravenous infusion of remifentanil and propofol.Then the patients received endotracheal intubation and mechanical ventilation.The end-tidal pressure of carbon dioxide was maintained at 30-35 mm Hg.Controlled hypotension was performed with nicardipine,and the mean arterial blood pressure was maintained at 50-70 mm Hg and heart rate at 60-90 bpm during operation.The patients received intravenous injection of flurbiprofen 50 mg 15 minutes before the end of operation for postoperative analgesia.When the visual analogue scale score was more than 3 during the first 6 hours after operation,flurbiprofen 50 mg was given intravenously as rescue analgesia.Results The incidence of rescue analgesia administered after operation was significantly greater in groups Ⅱ,Ⅲ and Ⅳ than in group Ⅰ,and greater in group Ⅳ than in groups Ⅱ and Ⅲ.There was no significant difference in the incidence of rescue analgesia administered during the first 6 hours after operation between groups Ⅱ and Ⅲ.Conclusion Preoperative sleep disturbance has adverse effects on the efficacy of flurbiprofen for postoperative analgesia in patients undergoing endoscopic nasal surgery.
7.Effect of sleep deprivation on cognitive function in rats undergoing propofol anesthesia
Hailiang DU ; Huan CHEN ; Ling PEI
Chinese Journal of Anesthesiology 2016;36(2):161-164
Objective To evaluate the effect of sleep deprivation on cognitive function in the rats undergoing propofol anesthesia.Methods Sixty healthy male Sprague Dawley rats,aged 14-18 weeks,weighing 200-250 g,were randomly assigned into 3 groups (n=20 each) using a random number table:control group (group C),propofol anesthesia group (group P),and sleep deprivation + propofol anesthesia group (group SDP).Propofol was given as a bolus of 15 mg/kg followed by an infusion of 40 mg · kg-1 · h-1 for 2 h in group P.After the rats were subjected to rapid eye movement sleep deprivation for 24 h,the rats received propofol anesthesia in group SDP.Before sleep deprivation,after sleep deprivation,and at 1,3 and 7 days after anesthesia,Morris water maze test was used to assess the learning and memory function,and the escape latency and frequency of crossing the original platform were recorded.Ten rats randomly selected from each group at 1 and 7 days after anesthesia were sacrificed,and brains were removed to observe the morphology of nerve cells in the hippocampal CA1 region (by Nissl's staining) and to detect the expression of phosphorylated Tau at Thr231 (Tau-pThr231) in the hippocampal CA1 region (by immunohistochemisty).Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was decreased,the expression of Tau-pThr231 in the hippocampal CA1 region was up-regulated at 1 day after anesthesia in P and SDP groups (P<0.05),especially in group SDP (P<0.05),and there was no significant difference between the groups at the other time points (P>0.05).The pathological changes were aggravated at 1 day after anesthesia in group SDP compared with group P,and there was no significant difference at 3 and 7 days after anesthesia between group SDP and group P.Conclusion Sleep deprivation can aggravate the transient cognitive dysfunction after propofol anesthesia,and the mechanism is related to promotion of Tau phosphorylation in the rats.
8.Inhibiting effects of ketamine on plasma TNF-? level and lung injury in septic shock rats
Ling PEI ; Dunke WANG ; Guozhong XU
Chinese Journal of Anesthesiology 1996;0(08):-
To investigate the inhibiting effects of ketamine on arterial plasma TNF-? concentration and lung injury in septic shock rat. Method: 40 Wister rats were divided into five groups. 15mg?kg~(-1) endotoxin (LPS) was intravenously injected alone (group Ⅰ)or ip ketamine 50,100 and 200mg?kg~(-1) before LPS, then ketamine was infused at 10mg?kg~(-1)?min~(-1) (Ⅱ, Ⅲ and Ⅳgroup). TNF-? was assessed with ELISA, and at the same time the arterial blood oxygen tension and lung water content were measured. Result: In contrast to normal control level, arterial plasma TNF-? levels and lung water content increased and arterial oxygen tension decreased after LPS in group Ⅰ, but in the rats of giving ketamine, plasma TNF-? level decreased more than that in the rats of giving LPS alone (group Ⅰ), change of arterial blood oxygen tension and lung water content in former groups were better than that of later, in dosage-dependent way. Conclusion: Ketamine can dose-relatedly decrease TNF-? concentration and lung injury degree induced by endotoxin.
9.Effects of pulmonary arterial endothelium cells injuryed by tumor necrosis factor ? on proliferation of pulmonary arterial smooth muscle cells and the interference effects of heat stress response
Ling PEI ; Junke WANG ; Wen FU
Chinese Journal of Anesthesiology 1994;0(01):-
Objective To investigate the effects of pulmonary arterial endothelium cells(PAEC) injuryed by tumor necrosis factor ? (TNF-?) on the proliferation of pulmonary arterial smooth muscle cells (PASMC) and the influence of heat stress response (HSR) on it .Methods Normal PAEC or PAEC endured with HSR were incubated with TNF-? at the concentrations of 500, 1000 and 2000u/ml in 1 hour, then cultured in DMEM without serum in 24 hours, the upper liquid was collected to prepare the endothelial cell-conditioned medium liquid (EC-CMⅠ or EC-CMⅡ ), in which PASMC were incubated in 24 hours as group Ⅰ or group Ⅱ respectively. The normal endothelial cell-conditioned medium liquid was also prepared to incubate PASMC in 24 hours as group Ⅲ.The PASMC were incubated without the endothelial cell-conditioned medium liquid as group Ⅳ.Flow cytometry was applied to determining the intracellular DNA content of the incubated PASMC. The fractions of different cytocycle phases were calculated according to the areas under the curves of DNA content.Results Compared with those of group Ⅳ, the percentage of PASMC in G0-G1 phases increased markedly ,and in S and G2-M phases decreased significantly in group Ⅲ (P
10.Neuromuscular effects and safety of an intravenous infusion of rapacuronium under inhalation or intravenous anesthesia
Wen FU ; Junke WANG ; Ling PEI
Chinese Journal of Anesthesiology 1994;0(04):-
0 05) The duration of T 1 recoving to 90% of baseline was 34 57min,and the recovery index was 24 29 min No any histamine related side effects were observed in all patients Conclusions Intravenous infusion of rapacuronium can produce safe and effective neuromuscular blockasde during desflurane, sevoflurane, isoflurane, or propofol anesthesia After the rapacuronium infusion of 45 60 min, the recovery from the neuromuscular blockasde is prolonged