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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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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 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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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 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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AIM: To investigate the effect of puerarin (Pur) on expression of Fas/FasL mRNA in lung tissue during pulmonary ischemia and reperfusion injury(PIRI) in rabbits.METHODS: Single lung ischemia and reperfusion animal model was used.The rabbits were randomly divided into three groups,sham operated group(sham,n=10),PIR group(I-R,n=30) and PIR+ Pur group(Pur,n=30).Changes of several parameters included apoptotic index(AI),wet to dry ratio of lung tissue weight(W/D) and index of quantitative assessment of histologic lung injury(IQA) were measured at 60,180 and 300 minutes after reperfusion in lung tissue.Meanwhile,the location and expression of Fas/FasL mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60,180,300 minutes after reperfusion.RESULTS: As compared with group I-R,Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia in group Pur.The values of AI,W/D and IQA showed significantly lower than that in group I-R at 60,180,300 minutes after reperfusion in lung tissue(P
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AIM:To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury,and to observe the effects of human thioredoxin (hTrx) on apoptosis in lung ischemia/reperfusion injury. METHODS:The single lung in situ ischemia/reperfusion animal model was used. Eighty four Wistar rats were randomly divided into control group (control),groups of ischemia for 1 h and reperfusion for different times (IR1h,IR3h,IR5h),and groups of IR + human thioredoxin treatment (IR1h + hTrx,IR3h + hTrx and IR5h + hTrx). Transmission electron microscope (TEM),terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and immunocytochemistry techniques were used to observe apoptosis,apoptosis signal-regulating kinase 1 (ASK1) and expression of Bcl-2 and Bax in various phases of lung ischemia/reperfusion. RESULTS:Cell apoptosis in lung tissues was significantly high,ASK1,Bcl -2 and Bax protein were upregulated in lung tissues of lung ischemia/reperfusion injury as compared to control (all P
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AIM:To investigate the effect of L-arginine(L-Arg) on expression of bcl-2,bax mRNA during pulmonary ischemia and reperfusion injury(PIRI) in rabbits.METHODS: Single lung ischemia and reperfusion animal model was used in vivo.The rabbits were randomly divided into three groups: sham operated group(sham,n=12),ischemia-reperfusion group(I/R,n=12) and I/R+ L-arginine group(L-Arg,n=12).Changes of several parameters,which included apoptotic index(AI),wet to dry ratio of lung tissue weight(W/D) and index of quantitative assessment of histologic lung injury(IQA),were measured at 300 min after reperfusion in lung tissue.Meanwhile the location and expression of bcl-2,bax mRNA as well as the ratio of bcl-2 mRNA/bax mRNA were observed.The lung tissue was prepared for light microscopic and electron microscopic observation at 60,180 and 300 min after reperfusion.RESULTS: As compared with I/R group,in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia,the expression of bcl-2 mRNA and the ratio of bcl-2 mRNA/bax mRNA were increased,and the expression of bax mRNA was decreased in L-Arg treatment group.The values of AI,W/D and IQA showed significantly lower than that in I/R group at 180 minutes after reperfusion in lung tissue(P