AIM:To investigate the regulatory role of retinoid X receptor(RXR)in oxidative stress response of rat type Ⅱ alveolar epithelial cells(AECII)induced by hypoxia/reoxygenation(HR).METHODS:The AECII were di-vided into control(C)group,HR group,HR+solvent dimethyl sulfoxide(DMSO)group(HD group),HR+RXR agonist 9-cis-retinoic acid(9-RA)group(RA group),and HR+RXR antagonist HX531 group(HX group).Cell Counting Kit-8(CCK-8)method was used to measure the cell viability.Immunofluorescence staining was used to detect the expression of surfactant protein A(SP-A)and RXRα in AECII.Kits were detected to the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in cells.Transmission electron microscopy was used to observe the ultrastructural changes of the cells.Western blot was used to detect the protein level of nuclear factor E2-related factor 2(Nrf2).RT-PCR was used to detect the expression level of Nrf2 mRNA.RESULTS:Compared with C group,the cell viability and SOD activity in HR,HD,RA and HX groups were decreased significantly(P<0.05),the MDA content were increased significantly(P<0.05),the Nrf2 mRNA and protein expression levels were decreased significantly(P<0.05 or P<0.01),and the immuno-fluorescence expression of RXRα was significantly increased(P<0.01).Compared with HR and HX groups,the cells in RA group showed significantly increased cell viability(P<0.05),increased SOD activity(P<0.05),decreased MDA con-tent(P<0.05),increased Nrf2 mRNA and protein expression levels(P<0.01),and significantly increased immunofluo-rescence expression of RXRα(P<0.01).CONCLUSION:Hypoxia/reoxygenation can aggravate the oxidative stress re-sponse of rat AECII,and RXR agonist intervention can alleviate HR-induced rat AECII injury by inhibiting oxidative stress.

AIM:To investigate the effects of sodium hydrosulfide(NaHS)on hexokinase 2(HK2)-nucleo-tide-binding oligomerization domain-like receptor protein 3(NLRP3)-gasdermin D(GSDMD)pathway and pyroptosis in-duced by lung ischemia/reperfusion(I/R)in rats.METHODS:Male Sprague-Dawley rats were divided into 6 groups:control group,control+NaHS group,I/R group,low-dose NaHS+I/R(L+I/R)group,medium-dose NaHS+I/R(M+I/R)group,and high-dose NaHS+I/R(H+I/R)group,with 6 rats in each group.The NaHS was administered via intraperi-toneal injection at 1.5 mL,30 min before modeling.The left lung tissues were collected 30 min after ischemia and 1 h af-ter reperfusion,and the wet/dry weight ratio and total lung water content were recorded.Hematoxylin-eosin(HE)staining was used to examine lung tissue morphological changes.The levels of malondialdehyde(MDA),myeloperoxidase(MPO)and lactate in lung tissues were measured with test kits.ELISA was employed to determine the levels of interleukin-1β(IL-1β)and IL-18.The expression of glycolysis-and pyroptosis-related indicators was analyzed by Western blot,qRT-PCR and immunofluorescence staining.RESULTS:Compared with control group,the rats in NaHS group showed no signifi-cant differences in all laboratory tests(P>0.05).The rats in I/R group exhibited significant lung injury,oxidative stress,increased lactate level,and up-regulated glycolysis and pyroptosis(P<0.05 or P<0.01).Compared with I/R group,the indicators in L+I/R group showed a downward trend(P<0.01)or no difference(P>0.05),while those in M+I/R group dis-played a significant reduction(P<0.05 or P<0.01).However,the indexes in H+I/R group exhibited no significant dif-ferences in these tests(all P>0.05).CONCLUSION:A moderate dose(56 μmol·L-1·kg-1)of NaHS mitigated the oc-currence of pyroptosis by inhibiting the HK2-NLRP3-GSDMD pathway,thus contributing to the attenuation of lung I/R in-jury in rats.

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.

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.

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.

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.

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.

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.

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.

[ 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.