Objective To study the effects of edaravone on the lung injury of severe acute pancreatitis （SAP） in rats. Methods Thirty-six SD rats were randomly divided into three groups: normal control group, model group and edaravone group, and SAP was induced by intraductal administration of 5% sodium taurocholate. Edaravone was given in edaravone group, while normal saline was given in normal control group and model group. After operation 6 h rats were executed, and dry/wet weight (D/W) ratio of lung was counted, and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity in serum and lung were detected, respectively. In addition, the levels of tumor necrosis factor-α (TNF-α), interleukin-1, -6 (IL-1, -6) of serum were detected.Results The MDA contentof serum and lung and the levels of TNF-α, IL-1, IL-6 in model group were markedly higher than those in normal control group and edaravone group, but D/W ratio of lung, SOD activity of serum and lung were significantly lower (Plt;0.05). Conclusion Edaravone can alleviate lung injury of rats caused by SAP.
Objective To explore the role of chronic ethanol ingestion in pulmonary fibrosis. Methods Twenty SD rats were randomly divided into a control group (n=10) and an ethanol group ( n=10) , and fed with quantitative non-ethanol and ethanol Lieber-DeCarli liquid diet every day respectively. All rats were sacrificed after 8 weeks. The morphological changes and collagen deposition of lung tissue were observed under light microscope by HE and Masson staining. Levels of glutathione (GSH) and hydroxyproline (HYP) in lung tissues were measured by colorimetric method. The content of connective tissue growth factor (CTGF) in lung tissue was detected by ELISA. Results Compared with the control group, varied degrees of alveolar and alveolar septal infiltration of inflammatory cells can be shown in the ethanol group, and also some alveolar wall damage or collapse.Masson staining showed that the ethanol group has more significant deposition of collagen fibers in alveolar interstitumthan the control group. The content of GSH in rat lung tissue reduced, but the contents of HYP and CTGF increased in the ethanol group compared with the control group [ GSH( mg/g) :0.08±0.04 vs. 0.22±0.14, HYP(mg/g) : 0.57±0.15 vs. 0.40 ± 0.09, CTGF(ng/mL) :306.57±46.86 vs. 134.02±79.82, Plt;0.05] . Conclusions Lieber-DeCarli ethanol liquid diet can establish a rat model of chronic ethanol ingestion. Lung injury and pulmonary fibrosis in rats can be induced by chronic ethanol ingestion. Ethanol may be one of the causes of the pulmonary fibrosis.
Objective To determine if mesenchymal stem cells ( MSCs) could be reconstructed as a vehicle for angiopoietin-1 ( Ang1) gene therapy in lung injury. Methods MSCs were obtained from adult male inbred mice and cultured to passage four. The cells were identified by fluorescence-activated cell sorting ( FACS) analysis and cell differentiation detection. Lentiviral vectors contained GFP and Ang1 gene were conducted in 293T cells through three plasmids co-transfection method. Then MSCs were transduced with Ang1 gene efficiently through lentiviral vectors. The mRNA expression of Ang1 in MSCs was detected by RT-PCR before and after transfection. Also fluorescence from MSCs was detected by fluorescence microscope every day after transfection. Two hours after LPS inhalation, mice were infused via jugular veinwith normal saline ( NS group) , lentiviral vector carrying Ang1 ( Ang1 group) , lentiviral vector carrying GFP ( MSCs group) , and lentiviral vector carrying Ang1 /GFP ( MSCs-Ang1 group) , respectively. Kaplan-Meier survival analysis was performed to compare the effects of MSCs-Ang1 on survival. And ectogenic MSCs origined lung cells were investigated in receipt mice. Results After passaged and purification,MSCs were confirmed to have the potential of differentiation. The lentiviral vectors carrying Ang1 and GFP were also identified. After transfection, the mRNA expression of Ang1 in MSCs was enhanced. Through the fluorescence microscope,MSCs get the most green fluorescence expression five days after the transfection when MOI was 20. Kaplan-Meier survival analysis showed that MSCs-Ang1 infusion had improved survival rates of lung injury rats compared with the control, but it did not reach statistical significance ( P = 0. 066) . Cells expressing GFP in lung tissues can be observed after MSCs were transplanted in vivo. Conclusions MSCs expressing Ang1 high can be constructed through lentiviral vector transfer, and MSCs-origined cells can be detected in receipt lungs after transplantation. So MSCs may serve as a vehicle for gene therapy in lung injury.
Objective To study the protective effects and mechanism of intermittent ventilation on lung injury during cardiopulmonary bypass(CPB). Methods Twenty-four patients with rheumatic heart disease (RHD) were divided into two groups with random number table: treatment group (n=13),given intermittent ventilation once every 5 minutes during CPB; control group (n=11),no ventilation during CPB. Blood samples were obtained preoperatively. A bronchoalveolar lavage was performed at 2 hours after CPB. The numbers of granulocytes, total protein (TP) and tumor necrosis factor-alpha(TNF-α) content in the bronchoalveolar lavage fluids(BALF) were measured, and lung oxygenate index (OI) were measured preoperatively and 1 hour, 4 hours after CPB termination,respectively. Results The numbers of granulocytes, TP and TNF-α content of treatment group in the BALF were significantly lower than those of the control group (Plt;0.01, P=0.02,0.02),and the lung OI of treatment group at 1 hour and 4 hours after CPB termination was also significantly lower than that of the control group(Plt;0.05); a significant increase of lung OI occurred in both groups at 1 hour and 4 hours after CPB when compared with the same group at baseline before CPB(Plt;0.05). Conclusion Intermittent ventilation has the protective effects on lung injury during CPB by decreasing granulocytes adhesion and alleviating lung inflammatory reaction and endothelial cells injury.
ObjectiveThe role of ferroptosis-related genes in the occurrence and development of lung injury caused by sepsis was investigated by bioinformatics methods, and the closely related genes were predicted. MethodsThe Dataset GSE154653 was downloaded from the gene expression database (GEO), and a total of 8 cases of microarray gene set were included in normal group and lipopolysaccharide (LPS)-induced sepsis lung tissue. The differential expression genes (DEGs) were screened out under conditions of |log2 FC|>1 and P.adj<0.05. Meanwhile, the selected DEGs were combined with the driver and suppressor genes of ferroptosis downloaded from the ferroptosis database (FerrDb) to obtain the differential genes associated with ferroptosis in sepsis (Fe-DEGs). These Fe-DEGs were further analyzed using R language, DAVID, and STRING online tools to identify GO-KEGG functions and pathways, and the construction of PPI network. Results The Bioinformatics approach screened out 3533 DEGs and intersected 53 key genes related to ferroptosis. The further biological process (BP) of GO enrichment analysis mainly involves the positive regulation of transcription, the positive regulation of RNA polymerase II promoter transcription, the cytokine mediated signaling pathway, and the positive regulation of angiogenesis. The molecular function (MF) mainly involves the same protein binding, transcriptional activation activity and REDOX enzyme activity. The pathways are enriched in iron death, HIF-1 signaling pathway and AGE-RAGE signaling pathway. Five key Fe-DEGs genes were screened by constructing PPI network, including CYBB, LCN2, HMOX1, TIMP1 and CDKN1A. Conclusion CYBB、LCN2、HMOX1、TIMP1 and CDKNIA genes may be key genes involved in ferroptosis of lung tissue caused by sepsis.
Objective To investigate the protective effects of endotoxin pretreatment on lung injury of rats with endotoxemia. Methods The rat model of acute endotoxemia was established by injecting lipopolysaccharide (LPS) intraperitoneally. Seventy-two male Wistar rats were randomly divided into three groups, ie. a saline control group (N, n=24) , a LPS-treated group (L, n=24) , and a LPS pretreated group ( P, n=24) . Each group was divided into 2 h, 4 h, 6 h, and 12 h subgroups. The rats in group P were firstly administered with introperitoneal injection of 0.25 mg/kg LPS. After 24 hours, they were subjected to the injection of 0.5 mg/kg LPS. The rats in group N and L received injection of equivalent amount of saline. After 72 hours, the rats in group L and P were challenged with intravenous injection of 10 mg/kg LPS, otherwise saline in group N. Six rats were killed at 2, 4, 6 and 12 hours respectively after injection of LPS in group L and P. The lungs were removed for detecting intercellular adhesion molecule-1 ( ICAM-1) , superoxide dismutase ( SOD) , and malondialdehyde (MDA) . Meanwhile the level of tumor necrosis factoralpha ( TNF-α) in serum was measured, and the pathological changes of lung were also examined. Results The contents of ICAM-1, MDA and TNF-α in the LPS-treated 4 h group were 75.07 ±0. 53, ( 3.93 ± 0.42) μmol/g, and (478.62 ±45.58) pg/mL respectively, significantly higher than those in the saline control group. The endotoxin pretreatment reduced the above indexes to 42.40 ±0.44, ( 2.89 ±0.49) μmol / g and ( 376.76 ±43.67) pg/mL respectively (Plt;0.05) . The content of SOD in the LPS-treated 4 h group was ( 6.26 ±0.31) U/mg, significantly lower than that in the saline control group. The endotoxin pretreatment increased SOD to ( 8.79 ±0.35) U/mg. Conclusion Endotoxin pretreatment can suppress the progress of lung injury in rats with endotoxemia and protect the lung tissue by down-regulating the inflammatory response and oxygen free radical production.
Objective To explore the migration and differentiation of bone marrow mesenchymal stem cells（MSCs） in lung . Methods MSCs were harvested from a male Wister rat. Sixty female Wister rats were randomly divided into four groups. The pulmonary fibrosis model was established by intratracheal instillation of bleomycin in group A-D. Immediately and 7 days after bleomycin administration respectively，the rats in group B and C received infusion with 5-bromodeoxynridine （BrdU） labeled MSCs via tail vein. And the rats in group D were infused MSCs without BrdU labeling serving as a negative control. The sry gene of Y chromosome was detected by polymerase chain reaction （PCR）. Double immunofluorescence staining was used to detected BrdU and surfactant associated protein-C （SP-C） expression in lung tissue，fresh bone marrow，and the 5th generation MSCs. Reverse transcriptipon-PCR was used to detect the expressions of SP-C mRNA and AQP-5 mRNA. Results The sry gene was detected in bleomycin induced lung injury tissues of the rats after MSCs infusion immediately and on the 7th day The MSCs in lung tissue could transformed into cells with ACEⅡ morphological features and molecular phenotype. The transformation rate was higher in the rats received MSCs infusion immediately than the rats received on 7th day. The 5th generation MSCs and fresh bone marrow expressed SP-C mRNA，without AQP-5 mRNA and SP-C expression. Conclusions Exogenous MSCs can be transplanted into injured lung tissues and transform into AECⅡ，especially in early stage of lung injury. The differentiation potential of MSCs can be activated in injury micro-environment.
Objective To investgate the expression of p38 mitogen-activated protein kinase （p38MAPK） in lung tissue of rats with severe acute pancreatitis （SAP）， and to explore the relationship between p38MAPK and pulmonary capillary barrier injury. Methods Forty male and healthy Sprague-Dawley （SD） rats were randomly （random number method） divided into sham operation （SO） group and SAP group， then rats of SAP group were sub-divided into 3， 6， 12， and 24 h group， each group enrolled 8 rats， respectively. SAP model rats were established by injecting 5% sodium taurocholate solution retrograde into the biliopancreatic duct. ELISA method was used to test the serum tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）， and pathological changes in lung and pancreas tissues were observed by HE staining. Immunohischemistry method was used to detect phosphorylated p38 （p-p38） protein and aquaporin 1 （AQP1） protein of lung tissues. The expression level of AQP1 mRNA was measured by quantitative real-time PCR. Results Hyperemia， edema， and inflammatory cell infiltration were observed in lung tissues， abundance of necrosis， part gland structure fuzzy or even disappear were observed in pancreas tissues of all 4 time point groups. Compared with SO group， levels of serum TNF-α and IL-1β were significantly higher in 4 time point groups （P＜0.05）. Lower expression level of p-p38 protein was detected in lung tissues of SO group， while in the early stage of SAP （SAP 3 h group）， the expression level of p-p38 protein significantly increased， which peaked in 6 h group and was still higher than SO group in 24 h group （P＜0.05）. Compared with SO group， the expression levels of AQP1 mRNA and protein were significantly lower in all 4 time point groups （P＜0.05）， which had negative correlation with the levels of serum TNF-α，IL-1β， and the expression level of p-p38 protein （r=-0.87， P＜0.05；r=-0.88， P＜0.05；r=-0.78， P＜0.05）. Conclusion The decrease of AQP1 protein in lung tissue is one of the vital causes for pulmonary capillary barrier injury in SAP， which probably works by the activation of p38MAPK and the excessive release of inflammatory cytokines.
Objective To explore the effects of exogenous ubiquitin on lung injury and serum nitric oxide ( NO) level of mice in the early stage of sepsis induced by cecal ligation and perforation ( CLP) . Methods Seventy-seven mice were devided randomly into three groups, ie. CLP Group ( n =28) , SHAM Group ( n =28) and UB Group( n = 21) . Six hours after operation, seven mice of both Group CLP and SHAM were sacrificed for lung and blood sampling. Meanwhile all mice of Group UB were injected intravenously with exogenous ubiquitin of 10 mg/kg body weight. Seven mice of each group were sacrificed at seven, eight and nine hours after operation. Lung water content and serum level of nitrate /nitrite ( NO2 /NO3 , the stable end-products of NO) , were determined. Pathological changes of lung were compared among the groups. Results Lung water content of Group UB was lower than that of Group CLP at each observational time point. Futhermore, there was a significant differentce in lung water content between Group UB and CLP at 9 hours. Pathological examination revealed that inflammatory hyperemia and infiltration of pulmonary parenchyma reduced in Group UB, compared with Group CLP. Serum NO levels of Group CLP and UB were similar at each time point. Conclusions In the early stage of murine sepsis induced by CLP, a single introvenous bolus of exogenous ubiquitin significantly decreases lung capillary vascular permeability, and probably makes a temporal reduction of acute lung injury while it has no obvious effects on serumNO level.