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AIM: To investigate the effect of dexmedetomidine (DEX) on renal injury induced by lung ischemia/reperfusion (I/R) in mice and its relationship with endoplasmic reticulum stress response.METHODS: Healthy SPF male C57BL/6J mice, weighing 20~24 g, aged 8~10 weeks, were randomly divided into 5 groups (n=10 each): sham operation group (sham group), I/R group, atipamezole (Atip) group, DEX group, and DEX+Atip group.In vivo lung I/R model was established by occlusion of the left pulmonary artery for 30 min followed by 180 min of reperfusion in the mice.The Atip (250 μg/kg), DEX (20 μg/kg) and DEX+Atip were intraperitoneally infused into the mice before left pulmonary hilus was blocked in Atip group, DEX group and DEX+Atip group, and other operations were the same as I/R group.After experiment, the mice were killed, and the renal tissues were harvested to observe the morphological changes.The enzymatic activity of caspase-3, serum creatinine and blood urea nitrogen, and cell apoptotic index of the renal cells were also analyzed.The expression of c-Jun N-terminal kinase (JNK), caspase-12, CCAAT/enhancer-binding protein homdogous protein (CHOP) and glucose-regulated protein 78 (GRP78) at mRNA and protein levels in the renal tissues was determined by RT-PCR and Western blot.RESULTS: Compared with sham group, the enzymatic activity of caspase-3, serum creatinine and blood urea nitrogen, renal cell apoptotic index, and the mRNA and protein levels of JNK, caspase-12, CHOP and GRP78 in I/R group were significantly increased (P<0.01), and the renal tissues had obvious damage under light microscope.Compared with I/R group, Atip group and DEX+Atip group, the enzymatic activity of caspase-3, serum creatinine and blood urea nitrogen, renal cell apoptotic index, and the mRNA and protein levels of JNK, caspase-12 and CHOP in DEX group were significantly decreased, and the expression level of GRP78 significantly increased (P<0.01).Furthermore, the renal tissue damage was obvious reduced.CONCLUSION: DEX effectively relieves the renal injury induced by lung I/R in mice, which may be associated with exciting α2-adrenergic receptor and inhibiting endoplasmic reticulum stress response.
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AIM:To investigate the effects and c-Jun N-terminal kinase(JNK) expression of dexmedetomidine (DEX) on A549 cells with hypoxia/reoxygenation (H/R) injury.METHODS:A549 cells were cultivated and were randomly divided into four groups (n =10):control group (N),DEX group (D),hypoxia/reoxygenation injury group (H),hypoxia/reoxygenation injury + DEX interfere group (HD).After all models were completed,the morphological changes of A549 cells were observed under the inverted microscope.Cell activity was detected by CCK-8 and the apoptosis index (AI) was detected by in situ end labeling (TUNEL) method.The expression of GRP78,p-JNK,caspase-3 at protein levels and GRP78,JNK mRNA were detected by Western blot and RT-PCR.RESULTS:Compared with N group,the number of adherent cells in H group decreased significantly,and cell morphology changed.The expression of OD value in H group decreased obviously (P ＜ 0.01),the expression of AI value,GRP78,p-JNK,caspase-3 protein and GRP78,JNK mRNA were significantly increased (P＜ 0.01).HD group compared with H group,the cell damage alleviated,the expression of OD value was increased (P ＜ 0.01),the number of apoptosis cells and the AI value in HD group were significantly decreased (P ＜ 0.01).Dramatically decreased the expression of p-JNK,caspase-3 protein and JNK mRNA (P ＜ 0.01).CONCLUSION:DEX can effectively alleviate A549 cells damage induced by H/R injury,which may be related to inhibition of the JNK pathway.
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Objective To observe the effects of Yiqi Huoxue Tongluo Jiedu fang (YHTJF) on pneumocyte apoptosis after lung ischemia/reperfusion (I/R) injury (LIRI) in mice and to investigate whether c-Jun N-terminal protein kinase (JNK) is involved in the mechanism of apoptosis.Methods Seventy C57BL/6J male mice were randomly divided into seven groups:normal control group (C group),carboxyl methyl cellulose-Na+normal control group (CMC-Na+C group),CMC-Na+sham group (CMC-Na+S group),CMC-Na+I/R group (CMC-Na+I/R group) and CMC-Na+YHTJF-low,-middle,-high dose groups (CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups).C group did not undergo any processing;in CMC-Na+S group,only was chest opened without clipping the lung hilum;in the rest of the four groups,they all underwent opening of the chest and clipping the lung hilum for 30 minutes,then the clipping of artery was relieved and left lung reperfusion was carried out for 3 hours.After operation,the mice were sacrificed,the lung tissues were harvested.Under light and electron microscopes,the lung morphological and ultra-structural changes were observed,and the changes of index of quantitative evaluation for alveolar damage (IQA) were determined.The terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was applied to evaluate the apoptosis index (AI) of the lung tissues.The protein and mRNA expressions of JNK and glucose regulating protein 78 (GRP78) in lung tissues were detected by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR);the correlations between lung AI and the expressions of mRNA and protein of JNK and GRP78,IQA were analyzed.Results Compared with CMC-Na+S group,IQA,AI and mRNA and the protein expressions of JNK and GRP78 in CMC-Na+I/R group were obviously higher [IQA:(74.00 ± 7.31)％ vs.(7.00 ± 1.23)％,AI:(64.40 ± 11.97)％ vs.(5.60 ± 1.14)％,JNK mRNA (gray value):1.143 ± 0.284 vs.0.152 ± 0.128,GRP78 mRNA (gray value):0.897 ± 0.129 vs.0.284 ± 0.044,JNK protein (A value):0.428 ± 0.074 vs.0.073 ± 0.052,GRP78 protein (A value):1.075 ± 0.145 vs.0.589 ± 0.060].Compared with CMC-Na+I/R group,the IQA,AI,protein and mRNA expressions of JNK and GRP78 in CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups were all lower,and the degree of reduction in group CMC-Na+YM was the most remarkable,greater than that in CMC-Na+YL or CMC-Na+YH group [IQA:(26.20 ± 3.35)％ vs.(34.00±5.34)％,(41.20±9.18)％,AI:(29.40±3.05)％ vs.(48.20±3.83)％,(39.20±6.14)％,JNK mRNA (gray value):0.681 ± 0.130 vs.0.804 ± 0.153,0.938 ± 0.11,GRP78 mRNA (gray value):0.450 ± 0.105 vs.0.747 ± 0.231,0.566 ± 0.115,JNK protein (A value):0.188 ± 0.049 vs.0.261 ± 0.065,0.209 ± 0.063,all P ＜ 0.01],compared with the CMC-Na+I/R group,the expression of GRP78 protein was obviously higher in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkably high was in CMC-Na+YH group (A value:1.429 ±0.226 vs.1.130±0.169,1.128 ±0.177,all P ＜ 0.01).The apoptosis of each group was mainly in the pulmonary vascular endothelial cells and alveolar epithelial cells,and brown particles were positive cells under light microscope.Under transmission electron microscope:nuclear pyknosis and margination under the nuclear membrane,cytoplasm condensed,lamellar bodies decreased and emptying increased,cell membrane microvilli decreased or disappeared,mitochondria swelling,inflammatory cells increased in alveolar septum and adhering onto the capillary walls could be seen in CMC-Na+I/R group.Compared with CMC-Na+I/R group,the lung tissue ultrastructural damage alleviated,ultrastructure of alveoli clearly seen,nuclear chromatin relatively uniform,cytoplasm increased,type Ⅱ alveolar epithelial cell surface microvilli relatively plenty,lamellar corpuscle number increased,mitochondria swelling ameliorated in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkable one was CMC-Na+YM group.AI was significantly positive correlated with the mRNA and protein expressions of JNK,GRP78 and IQA (r =0.907,0.928,0.880,0.712,0.911,all P ＜ 0.01).Conclusions YHTJF may effectively alleviate the cell apoptosis in mice LIRI,and its mechanism may be related to the inhibition of JNK pathway.
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Objective To evaluate dexmedetomidine-induced cardioprotection in a mouse model of lung ischemia-reperfusion (I/R) and the relationship with endoplasmic reticulum stress.Methods Forty healthy SPF male C57BL/6J mice,weighing 20-24 g,aged 8-10 weeks,were divided into 4 groups (n=10 each) using a random number table:sham operation group (Sham group),lung I/R group (I/R group),dexmedetomidine group (Dex group) and dexmedetomidine plus atipamezole (specific α2-adrenergic receptor antagonist) group (DA group).The model of lung I/R injury was established by clamping the left hilum of lung for 30 min followed by 180 min of reperfusion.In group Sham,only sternotomy was performed,the hilum of lung was not clamped,and the mice were mechanically ventilated for 210 min.In Dex and DA groups,dexmedetomidine 20 μg/kg and dexmedetomidine 20 μg/kg plus atipamezole 250 μg/kg were injected intraperitoneally,respectively,at 30 min before establishment of the model.At 180 min of reperfusion,blood samples were collected from the orbit for determination of creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) activities in serum.The animals were then sacrificed,and hearts were removed for determination of apoptosis in cardiomyocytes (by TUNEL) and expression of phosphorylated c-Jun N-terminal kinase (p-JNK),caspase-12,CCAAT/enhancer-binding protein homologous protein (CHOP) and glucose-regulated protein 78 (GRP78) in myocardial tissues (by Western blot),and expression of JNK,caspase-12,CHOP,GRP78 mRNA in myocardial tissues (by real-time polymerase chain reaction).Apoptosis index was calculated.Results Compared with Sham group,the serum CKMB and LDH activities and apoptosis index were significantly increased,the expression of p-JNK,JNK mRNA,and caspase-12,CHOP and GRP78 protein and mRNA was up-regulated in I/R,Dex and DA groups (P＜0.01).Compared with I/R group,the serum CK-MB and LDH activities and apoptosis index were significantly decreased,the expression of p-JNK,JNK mRNA,and caspase-12 and CHOP protein and mRNA was down-regulated,the expression of GRP78 protein and mRNA was up-regulated in group Dex,and the expression of GRP78 protein and mRNA was significantly up-regulated (P＜0.01),and no significant change was found in the other parameters in group DA (P＞0.05).Compared with DEX group,the serum CK-MB and LDH activities and apoptosis index were significantly increased,the expression of pJNK,JNK mRNA,and caspase-12 and CHOP protein and mRNA was up-regulated (P＜0.01),and no significant change was found in the expression of GRP78 protein and mRNA in DA group (P＞0.05).Conclusion Dexmedetomidine can reduce myocardial injury induced by lung I/R,and the mechanism may be related to activation of α2-adrenergic receptors and inhibition of endoplasmic reticulum stress in myocardial cells of mice.
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Objective To investigate the effect of dexmedetomidine (DEX) on the reduction of brain injury induced by lung ischemia/reperfusion (I/R) in mices through inhibiting excessive endoplasmic reticulum stress response (ERS).Methods Fifty healthy SPF male C57BL/6J mices,weighing 20-24 g,aged 8-10 weeks,were divided into 5 groups (n =10 each) using a random number table:sham operation group (sham group),lung ischemia/reperfusion group (I/R group),atipamezole goup (Atip group),dexmedetomidine group (DEX group),dexmedetomidine plus atipamezole group (DA group).The model of lung I/R injury was established by clamping the left hilum of lung for 30 min followed by reperfusion for 180 min.In Atip,DEX and DA groups,atipamezole 250 μg/kg,dexmedetomidine 20 μg/kg and dexmedetomidine 20 μg/kg plus atipamezole 250 μg/kg were injected intraperitoneally,respectively,at 30 min before modeling,other procedures were as the same as the I/R group.At 180 min of reperfusion,the animals were sacrificed and the brain tissues were harvested for the observation of morphological changes.The Caspase-3 activity and the apoptosis index of the brain cells were also determined.The levels of protein and mRNA expression of p-JNK,Caspase-12,CHOP and GRP78 in brain tissues were detected by Western blot and RT-PCR.The datas were analyzed using SPSS 19.0 software and multiple-group comparisons were performed using one-way ANOVA,and P ＜ 0.05 for the difference was statistically significant.Results Compared with the sham group,the Caspase3 activity and brain cell apoptosis index,the protein levels and mRNA expressions of p-JNK,Caspase12,CHOP,GRP78 were significantly increased (P ＜ 0.01),brain tissues had obvious damage in I/R,Atip,DEX and DA groups;compared with I/R,Atip and DA group,brain tissues damage was obvious reduced in DEX group,and the Caspase3 activity,brain cell apoptosis index,the protein levels and mRNA expression of p-JNK,Caspase12,CHOP in DEX group were significantly lower,and GRP78 expression increased significantly (P ＜ 0.01).Comparisons among I/R,Atip and DA groups,there were no significant differences in degree of brain injury,Caspase3 activity,brain cell apoptosis index,the protein levels and mRNA expressions of p-JNK,Caspase12,CHOP (P ＞ 0.05),while the expression of GRP78 in DA group was significantly increased (P ＜ 0.01).Conclusion DEX can effectively relieve the brain injury induced by lung I/R in mice,which may be associated with stimulation of α2 adrenergic receptor and inhibition of excessive endoplasmic reticulum stress response and reducing brain cell apoptosis.
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Objective To evaluate the role of CCAAT/enhancer-binding protein homologous protein (CHOP) in ischemia-reperfusion induced lung injury, and clarify the potential molecular mechanism.Methods Fifty mice of C57BL/6J were randomly divided into five groups, 10 mice in each group, including Sham operation group(sham group),ischemia/reperfusion group (I/R group), I/R +PBS+Lipofectamine group(I/R+PBS+Lipo group) , I/R+scramble siRNA group( I/R+siRNASCR group) , I/R+CHOPsiRNA group( I/R+siRNACHOP group).The content of total lung water (TLW), wet-to-dry weight ratio (W/D) of lung and index of quantitative assessment of histologic lung injury (IQA) were detected, CHOP mRNA and protein expression were detected by RT-PCR and Western blot.Results Compared with Sham group, W/D, TLW and IQA were significantly elevated in I/R group,I/R+PBS+Lipo group and I/R+siRNASCR group (P<0.05).Moreover, the W/D,TLW and IQA reduced with CHOP-siRNA treatment, respectively(P<0.01).Compared with Sham group, CHOP mRNA and protein expressions were significantly elevated in I/R group,I/R+PBS+Lipo group and I/R+siRNASCR group, Moreover, the CHOP mRNA and protein expressions reduced with CHOP-siRNA treatment, respectively(P<0.01). Conclusion I/R could cause excessive unfolded protein response in lung tissue, and induce apoptosis by CHOP signal pathway, damage lung tissue. The inhibition of CHOP pathway could alleviate lung ischemia-reperfusion injury.
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[ ABSTRACT] AIM:To explore the effect of dexmedetomidine ( DEX) on the CCAAT/enhancer-binding protein-homologous protein ( CHOP) pathway during lung ischemia-reperfusion ( I/R) in mice.METHODS:C57BL/6J male mice were randomly divided into sham operation group ( sham group) , lung ischemia/reperfusion group ( I/R group) , ischemia/reperfusion +normal saline group ( I/R+NS group ) and ischemia/reperfusion+dexmedetomidine group ( I/R+DEX group) .Dexmedetomidine was infused intraperitoneally with 25 μg/kg for 30 min prior to the ischemia period in I/R+DEX group, the normal saline was administrated with the same volume of dexmedetomidine in I/R+NS group.After fini-shed the 3 h-reperfusion period , the left lung tissues were harvested to determine lung wet/dry weight ( W/D) , the total lung water content ( TLW) , and index of quantitative evaluation for alveolar damage ( IQA) .Morphological observation and terminal-deoxynucleotidyl transferase mediated nick end labeling ( TUNEL) were applied to evaluate the structure changes and the apoptosis index (AI) of the lung tissues.The expression of CHOP and glucose-regulated protein 78 (GRP78) at mRNA and protein levels in the lung tissues was detected by Western blot and RT-PCR.RESULTS:Compared with sham group, the W/D, TLW, IQA, AI, the mRNA and protein expression of CHOP and GRP78 obviously increased, and the left lung tissues structure were damaged more obviously both in I/R group and I/R+NS group.Compared with I/R group, the W/D, TLW, IQA, AI and the protein and mRNA expression of CHOP in I/R+DEX group decreased, the injury of the left lung tissue structures induced by I/R in I/R+DEX group were also alleviated .CONCLUSION:DEX alleviates the lung I/R injury, which may be related to inhibition of apoptosis mediated by CHOP pathway.
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Objective To observe the clinical effect of therapy of budesonide/formoterol powder for inhalation in treatment of adult with bronchial asthma.Methods 90 cases with bronchial asthma and were divided into observation group and control group randomly from February 2014 to February 2015.45 cases in each group.Control group was treated with budesonide inhalants +formoterol inhalants, observation group was given budesonide/formoterol powder inhalation.Changes of related indicators were followed up and recorded.Results After treatment, IL-5, IL-12 and IFN-γwere (60.2 ±9.7)pg/mL,(31.4 ±3.1)pg/mL,(1.6 ±0.2) ng/mL of observation group were better than control group (72.8 ±10.7)pg/mL,(38.5 ± 5.6)pg/mL,(2.3 ±0.3) ng/mL (P<0.05).After treatment,total effective rate in observation group was 68.9%, which was better than control group (48.9%)(P<0.05).Conclusion Clinical effect of budesonide/formoterol powder for inhalation in treatment of patients with bronchial asthma is accurate, and there is no obvious adverse reaction.
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Objective To evaluate the effect of small interfering RNA targeting caspase-12 (caspase-12-siRNA) pretreatment on lung ischemia/reperfusion (I/R) injury in mice.Methods Forty male C57BL/6J mice,aged 6-8 weeks,weighing 16-24 g,were randomly allocated into 4 groups (n =10 each) using a random number table:sham operation group (group S),group I/R,negative control group (group NC) and caspase-12-siRNA pretreatment group (group siRNA).Lung I/R was induced by clamping the left pulmonary hilum for 30 min followed by 3 h reperfusion in anesthetized mice in IR,NC and siRNA groups.At 48 h before ischemia,negative control siRNA 20 μg and caspase-12-siRNA 20 μg were instilled intranasally in NC and siRNA groups,respectively,and the total volume was 50 μl.At 3 h of reperfusion,the animals were sacrificed and the left lung was removed for determination of wet/dry lung weight (W/D) ratio and lung water content in lung tissues and for microscopic examination.Pulmonary ultrastructure was examined with electron microscope.The quantitative evaluation index (QEI) for alveolar damage and apoptosis rate were calculated.Results Compared with group S,W/D ratio,lung water content,QEI for alveolar damage and apoptosis index were significantly increased in IR and NC groups,QEI for alveolar damage and apoptosis index were increased in group siRNA (P ＜ 0.05).Compared with IR and NC groups,W/D ratio,lung water content,QEI for alveolar damage and apoptosis index were significantly decreased (P ＜ 0.05),and the pathological changes of lungs were alleviated in group siRNA.There was no significant difference in the indices mentioned above between groups IR and NC (P ＞ 0.05).Conclusion Caspase-12-siRNA pretreatment can attenuate lung I/R injury in mice.
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AIM:To investigate the effects of voltage-dependent K +channel 1.5 (Kv1.5) on the proliferation and apoptosis of rat pulmonary artery smooth muscle cells (PASMCs) under hypoxia+hypercapnia condition and the relation-ship with mitogen-activated protein kinase(MAPK) signal pathway.METHODS:The PASMCs isolated from the male SD rat were cultured under hypoxia +hypercapnia condition, and randomly divided into normal group (N group), hypoxia+hyper-capnia group (HH group), hypoxia+hypercapnia+DMSO incubation group (HD group), hypoxia+hypercapnia +U0126 (an extracellular signal-regulated kinase 1/2 inhibitor) incubation group (HU group), hypoxia+hypercapnia+SB203580 (a p38 mitogen-activated protein kinase inhibitor ) incubation group (HS group), and hypoxia+hypercapnia+anisomycin (an agonist of MAPK) incubation group (HA group).Cell Counting Kit-8 was used to detect the cell viability.The protein expression of Kv1.5, PCNA and Bax was detected by Western blotting .RESULTS:Compared with N group , the cell via-bility and PCNA protein expression in HH group and HD group were significantly raised (P0.05 ) .Compared with HD group , the cell viability and PCNA protein expression in HU group , HS group and HA group were decreased (P<0.05 or P<0.01), but Kv1.5 protein and Bax protein were raised (P<0.01), with the most significant changes in HA group .CONCLUSION:The regulation of Kv1.5 to the proliferation and apoptosis of PASMCs under hy-poxia+hypercapnia condition might have a relationship with the activation of MAPK signal pathway .
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Objective To observe the changes of cell apoptosis and levels of Bcl-2 and Bax protein in myocardium after cardiopulmonary resuscitation (CPR) in rats and to study the protective effects of different doses of exogenous phosphocreatine (creatine phosphate, CP) on cell apoptosis. Methods A total of 32 male adult SD rats were randomly divided into 4 groups, namely control group ( group A), CPR group (group B), low dose CP group (group C, phosphocreatine 0. 5 g/kg given at beginning of CPR and 1.0 g/ kg 2 hour after CPR) and high dose CP group ( group D, phosphocreatine 1.0 g/kg at beginning of CPR and 2. 0 g/kg 2 hours after CPR) . Cardiac arrest was induced by asphyxiation and CPR was started 7 min after asphyxiation it groups B, C and D. Myocardium samples were taken 24 hours after CPR for detecting myocardium cell apoptosis by TUNEL method. The levels of Bcl-2 and Bax protein were measured by using immunohistochemistry. Experimental data were processed with variance analysis in SPSS package. Results Compared with group A, myocardium cell apoptosis index (AI), and the levels of Bcl-2 and Bax proteinincreased significantly in groups B, C and D (P ＜0. 01 ), and Bcl-2/Bax ratio decreased significantly (P ＜0. 01 ) . Compared with group B, myocardium cell AI and levels of Bcl-2 and Bax protein decreased significantly in groups C and D ( P ＜ 0. 01 ), and Bcl-2/Bax ratio increased significantly ( P ＜ 0. 01 ) .Compared with group C, myocardium cell AI and levels of Bcl-2 and Bax decreased significantly in group D (P ＜ 0. 05 ), and Bcl-2/Bax ratio increased significantly ( P ＜ 0. 05 ) . Conclusion Exogenous phosphocreatine, especially inlarge dose, could inhibit apoptosis of myocardium cells and alleviate myocardium injury after CPR in rats.
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Objective To study the effect of Hemin on the level of neuroglobin (NGB) in cerebral cortex,neurodeficit score (NDS) and pathological changes in cerebral cortex after cardiopulmonary resuscitation (CPR) in rats. Method A total of 120 male Sprague Dawley(SD) rats were divided randomly into control group(A), CPR group(B) and Hemin group(C). The animal model of cardiac arrest (CA) induced by asphyxia was established and CPR was performed. The NGB level in cerebral cortex, NDS and pathological changes in cerebral cortex were examined 0.5 h,3 h,6 h, 12 h, 24 h after restoration of spontaneous circulation (ROSC) in each group. Experimental data were analyzed by using one-factor analysis of variance and Tukey test. Results In comparison with group A, the levels of NGB were significantly higher 12 h,and 24 h after ROSC (P ＜0.05 and P ＜0.01), the values of NDS were significantly lower at each interval after ROSC ( P ＜ 0.01 ) ,and the pathological changes were more severe at each interval after ROSC in group B. In comparison with group A, the levels of NGB were significantly higher 6 h, 12 h and 24 h after ROSC ( P ＜ 0.05 and P ＜ 0.01 ), and the values of NDS were significantly lower 3 h, 6 h, 12 h after ROSC ( P ＜ 0.01) in group C. In comparison with group B, the levels of NGB were significantly higher 12 h and 24 h after ROSC, the values of NDS were significantly higher 12 h and 24 h after ROSC, and the pathological changes were less in group C. Conclusions The NGB level increased in cerebral cortex, the NDS level decreased and severe pathological changes occurred in rats after CPR. The hemin treatment up-regulated the level of NGB, improved the NDS, mitigated pathological changes, alleviating the cerebral injury after CPR.
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AIM: To investigate the effects of non-invasive ischemic preconditioning on nitric oxide (NO)/endothelin-1 (ET-1) imbalance and gas exchange impairment following limb ischemia reperfusion in patients undergoing unilateral lower extremity surgery with tourniquet. METHODS: Twenty-seven patients aged 25-65 years, whose tourniquets duration varied from 1 h to 1.5 h and matched American Society of Anesthesiologists Physical Status Ⅰ-Ⅱ, were randomized into two groups: a control group (n=14) and a ischemic preconditioning group (IPC group, n=13) in which patients received three cycles of 5 min of ischemia/5 min of reperfusion before tourniquet inflation. Radial arterial blood gas, plasma malondialdehyde (MDA) and NO, serum ET-1 and interleukin-6 (IL-6) were measured just before tourniquet inflation(T_0), 1 h after inflation(T_1), and 0.5 h(T_2), 2 h(T_3), 6 h(T_4), 24 h(T_5) after tourniquet deflation. Meanwhile NO/ET-1 ratio, alveolar-arterial oxygen gradient (P_(A-a)DO_2) and intrapulmonary shunt (Qs/Qt) were calculated. RESULTS: In control group, arterial partial pressure of oxygen (PaO_2) were decreased, while P_(A-a)DO_2 and Qs/Qt were increased significantly at T_4 compared to the baselines at T_0 (P<0.01). Plasma NO levels and NO/ET-1 ratios decreased gradually after tourniquets deflation and statistical significances were observed at T_3 (P<0.01) with a valley at T_4 (P<0.01) and recovered to baselines at T_5. Serum ET-1, IL-6 and plasma MDA began to increase remarkably after T_3 (P<0.05 or P<0.01), peaked at T_4 and dropped slightly at T_5. The changes above-mentioned could be well attenuated by the application of IPC (P<0.05 or P<0.01) except PaO_2 (P>0.05). CONCLUSION: Clinical application of unilateral tourniquet within safe time limit (1.5 h) may lead to limb ischemia reperfusion and further pulmonary gas exchange impairment, which could be partially attenuated by the application of IPC via alleviating NO/ET-1 imbalance.
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AIM: To investigate the effect of ligustrazine on hydrogen sulfide (H_2S) system in pulmonary hypertension induced by hypoxic hypercapnia in rats. METHODS: Thirty Sprague-Dawley rats were randomly divided into 3 groups: control group (C), hypoxic hypercapnia group (HH), and hypoxic hypercapnia+ligustrazine group (HH+L). The change of hemodynamics was measured. The ratio of vessel wall area and total area of arteriae pulmonalis were observed under light microscope. The apoptosis of arteriae pulmonalis was tested with TdT-mediated dUTP nick end labeling (TUNEL), and the apoptosis index was calculated. Plasma level of hydrogen sulfide and activity of hydrogen sulfide generating enzymes in homogenates of rat lung tissue were evaluated by sensitive modified sulfide electrode method. Cystathionine-γ-lyase (CSE) mRNA in lung tissues was determined by RT-PCR. RESULTS: The level of mean pulmonary arterial pressure, the ratio of vessel wall area/total area and the right ventricle/left ventricle+septum were significantly higher in HH group than those in C group, and the value was obviously lower in HH+LTZ group than that in HH group (all P<0.01). The mean carotid arterial pressure of 3 groups had no significant difference (P>0.05). The apoptotic index of arteriae pulmonalis in HH group and HH+LTZ group was significantly lower than that in C group, and that in HH+LTZ group was significantly higher than that in HH group (P<0.05 or P<0.01). Plasma level of H_2S, the activity of H_2S generating enzymes in homogenates of rat lung tissue, cystathionine-γ-lyase (CSE) mRNA in lung tissues in HH group were significantly lower than those in C group (all P<0.01), and those in HH+LTZ group were significantly lower than those in HH group (all P<0.01). CONCLUSION: Ligustrazine up-regulates the expression of cystanthionine-γ-splitting enzyme (CSE), enhances the activity of CSE and increases the level of H_2S to prevent pulmonary hypertension induced by hypoxic hypercapnia.
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Objective To investigate the serum concentration and expressions of S100β protein in hippocampus CA1 region and the changes of water content in rats with asphyxia following ulinastatin injection after cardiopul-monary resuscitation (CPR). Method One hundred twenty male adult SD rots were randomly divided into 3 groups:sham-operation group, CPR group and ulinastafin group. And each group was further divided into 5 sub-groups (n=8) based on various intervals, 0.5 h,3 h,6 h,12 h and 24 h after tracheotomy in sham-operation group or after ROSC in CPR group and ulinastatin group. Asphyxial cardiac arrest and CPR model of rat was used in CPR group and ulinastatin group in which bolus dose of 100 000 U/kg ulinastatin was injection into arteria carotis. Anaesthesia, tracheotomy and vascular canratlafion without asphyxia and CPR in sham-operation group. Samples from subgroups were taken at different intervals. Brain water content was measured by using wet-dry weight method. Serum S100β protein was measured with enzyme-linked immunosorbent assay ( ELISA). The expres-sion of S100β protein in hippocampus CA1 region was measured by using immunohistochemistry. Data were ana-lyzed by SPSS version 10.0 software. Results Compared with sham-operation group, the brain water content of rats elevated significantly in all CPR subgroups after ROSC (P<0.05 or P<0.01). The brain water content of rats decreased significantly 12 h and 24 h after ROSC in ulinastatin group in comparison with CPR group (P<0.05). The serum S100β protein started to elevated significantly 0.5 h after ROSC in CPR group, and reached the peak 12 h after ROSC (P<0.01).serum S100β decreased 6 h,12 h and 24 h after ROSC in ulinastatin group compared with CPR group (P<0.01).The expression of S100β protein in hippocampus CA1 region remained at a low level in sham-operation group. The expression of S100β protein elevated significantly in all CPR subgroups after ROSC compared with sham-operation group (P<0.05 or P<0.01). Compared with CPR group, the ex-pression of S100β protein decreased after ROSC in ulinastatin group(P<0.05) .However,the expression of S100β protein in hippocampus CA1 region was significantly correlative with brain edema in all subgroups of CPR (r=0.862, P<0.05). Conclusions Ulinastatin can decrease serum S100β protein and the expression of S100β pro-tein in hippocampus CA1 region and lessen the severity of cerebral edema, alleviate the brain isehemic injury in rats after cardiopulmonary resuscitation.
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Objective To investigate the influence of injection carthamus tinctorius D. (1C) on the expression of intercellular adhesion molecule-1(ICAM-1) during the ischemia-reperfusion injury of lung (LIRI) in rabbits and its potential mechanism. Method Single lung ischemia-reperfusion animal model was induced in rabbits. A total of 30 Japanese white rabbits were randomly divided into sham-operation group (S group, n =10), ischemia-reperfusion group (I/R group, re = 10) and ischemia-reperfusion plus 1C group (1C group, n = 10) .The rabbits of 1C group received 1C 2.0 ml/kg injected intravenously just at 20 min before ligation of artery involved and the same dose of 1C instantly at the initiation of reperfusion. Malondialdehyde (MDA) , superoxide dismutase (SOD) and xanthine oxidase(XO) in serum were measured. The lung tissue was sampled and assayed wet/dry weight ratio (W/D), contents of myeloperoxidase (MPO) at the end of the experiment, and ultrastructure changes were observed under electron microscope. The expression of ICAM-1 was measured by using immunohistochemistry(IHC) . snd one-way ANOVA was used for statistical analysis. Results In I/R group, serum XO and MDA increased and SOD decreased, whereas the same pattern of changes but less magnitude happened in 1C group ( P < 0.01). The values of W/D and MPO were much higher in I/R group, but lower in 1C group. Under electron microscope, the ultrastructure of lung tissue showed pathological changes in the rabbits of I/R group,and these changes were greatly attenuated in the rabbits of 1C group . The IHC showed that ICAM - 1 in lung tissue of I / R group was (2.94±0.48) which was significantly higher than that of 1C group(1.75 (P < 0.01). Conclusions Injection Carthamus tinctorius D. may meliorate the ischemia-reperfusion injury of lung by way of suppressing the expression of ICAM-1, inhibiting neutrophil aggregation, lowering oxygen free radical level and decreasing lipid peroxidation.
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Objective To study the effect of ligustrazini (LGT) and propofol (PRO) on hepatocellular energy metabolism in the reperfusion injury after hepatic ischemia in rabbits and its mechanism. Methods The rabbits were randomly divided into four groups (n=10 in each), hepatic ischemia-reperfusion (HIR) group, HIR+LGT group (B), HIR+PRO group (C) and HIR+ LGT+PRO group (D). The contents of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), total adenylic acid number (TAN), energy charge (EC), malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide products (NO2-/NO3-) in the liver tissue were measured at 45 minutes after reperfusion. Results The contents of ATP, NO and SOD activity of the liver tissue in B, C, D groups were higher than those in HIR group (P
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Objective To investigate the role of HO-1 in the protection of rat heart from anoxia/reoxygenation induced injury and its underlying mechanism.Methods LVEDP,LVDP and dp/dtmax were analyzed by the Langendorff method in isolated rat heart.Lactate dehydrogenase(LDH), infarct area,COHb and 6-keto-PGF1? were further determined in the experiment.Results After intraperitoneal injection of HO-1 inducer hemin,CO concentration in rat blood enhanced(P
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Objective To investigate the change of caspase-3 in rabbits after lung ischemia-reperfusion injury(LIRI) and the effect of puerarin(葛根素).Methods Thirty healthy rabbits used for unilateral lung ischemia-reperfusion model were randomly divided into 3 groups(each n=10): control group(C group),lung ischemia-reperfusion group(I/R group) and puerarin group.The activity of serum superoxide dismutase(SOD),the contents of serum malondialdehyde(MDA) and nitric oxide(NO),the wet to dry weight(W/D) ratio of lung tissue and the index of quantitative assessment of histological lung injury(IQA) were measured respectively in different groups;the pneumocyte apoptosis index(AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL);caspase-3 protein and mRNA expression were studied by using in situ hybridization(ISH) and immunocytochemistry(IHC) techniques in the groups mentioned above.Results The activity of SOD and content of NO were significantly lower in I/R group than those in C group(both P
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The changes of tumor necrosis factor-alpha (TNF-alpha) and brain ultrastructure during cardiopulmonary resuscitation and the effects of ulinastation injection were observed, and the mechanism was investigated. Twenty-four adult healthy Sprague-Dawley rats were randomly divided into control group (8 rats), resuscitation group (8 rats) and ulinastatin (UTI) group (8 rats). Rats in control group underwent tracheotomy without clipping the trachea to induce circulatory and respiratory standstill. Rats in resuscitation and ulinastatin group were subjected to the procedure of establishing the model of cardiopulmonary cerebral resuscitation (CPCR). Rats in ulinastatin group were given with UTI 104 U/kg once after CPCR. In the control group, the plasma was collected immediate, 30 min, 2 h, 4 h, and 6 h after tracheotomy. In resuscitation group and UTI group, plasma was collected immediate after tracheotomy, 30 min, 2 h, 4 h and 6 h after successful resuscitation. The plasma levels of TNF-alpha were determined by radioimmunoassay (RIA). At the end of the experiment, 2 rats were randomly selected from each group and were decapitated. The cortex of the brain was taken out immediately to observe the ultrastructure changes. In control group, there were no significant differences in the level of TNF-alpha among different time points (P>0.05). In resuscitation group, the level of TNF-alpha was increased obviously after resuscitation (P<0.01) and reached its peak 2 h later after resuscitation. An increasing trend of TNF-alpha showed in UTI group. There were no differences in TNF-alpha among each sample taken after successful resuscitation and that after tracheotomy. The utrastructure of brains showed the injury in UTI group was ameliorated as compared with that in resuscitation group. In early period of CPCR, TNF-alpha was expressed rapidly and kept increasing. It indicated that TNF-alpha might take part in the tissue injury after CPCR. The administration of UTI during CACR could depress TNF-alpha and ameliorate brain injury. By regulating the expression of damaging mediator, UTI might provide a protective effect on the tissue injury after CPCR.