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Objective Optimizing the preparation method to improve the quality of mouse lung cryosections to help enhance the specificity of immunofluorescence staining and obtain more accurate and reliable experimental result.Methods C57BL/6 mouse lung tissue was used to make cryosections via the traditional post-freezing fixation method,pre-freezing fixation method,and a modified perfusion pre-freezing fixation method.A laser-scanning confocal microscope was used to observe lung tissue immunofluorescence staining.Whole areas of mouse lung slices were scanned by fluorescence microscope,and the numbers of intact airways per unit area of lung tissue were calculated.Results In the lung cryosections made via the traditional post-freezing fixation method,the alveoli structure was damaged,the airway wall was seriously disrupted,and there was non-specific staining.Lung cryosections made via the pre-freezing fixation method showed relatively intact alveolar and airway structures but collapsed alveoli and several destroyed airways.In the lung cryosections obtained via the modified perfusion pre-freezing fixation method,the structure and morphology of the alveoli and airways were intact and clear.Additionally,the locations of multiple proteins targeted with immunofluorescence staining were accurate.The number of intact airways(diameter ≥100 μm)per unit area in the lung cryosections obtained via the modified perfusion pre-freezing fixation method was higher than that from cryosections made using the pre-freezing fixation method((0.66±0.15)/mm2 vs(0.33±0.14)/mm2,P＜0.05)and was also significantly higher than that from sections made using the traditional post-freezing fixation method((0.66±0.15)/mm2 vs(0.02±0.04)/mm2,P＜0.01).Conclusions The modified perfusion pre-freezing fixation method for cryosections is conducive to maintaining the integrity of mouse lung tissue morphology and obtaining high-quality multiplex immunofluorescence staining result.
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Relevant research data shows that there is a certain degree of energy metabolism imbalance in highland residents. Protein phosphatase 4 (PP4) has been found as a new factor in the regulation of sugar and lipid metabolism. Here, we investigate the differential expression of PP4 at a simulated altitude of 4,500 m in the heart, lung, and brain tissues of rats. A hypoxic plateau rat model was established using an animal decompression chamber. A blood routine test was performed by an animal blood cell analyzer on rats cultured for different hypoxia periods at 4,500 m above sea level. Quantitative polymerase chain reaction (qPCR) and western blot were used to detect the changes of protein phosphatase 4 catalytic subunit (PP4C) gene and protein in heart, lung, and brain tissues. The PP4C gene with the highest expression level found in rats slowly entering the high altitude area (20 m-2200 m-7 d-4500 m-3 d) was about twice as high as the low elevation group (20 m above sea level). The simulated high-altitude hypoxia induced an increase of PP4C expression level in all tissues, and the expression in the lung tissue was twice as expressed as heart and brain tissue at high altitude (P < 0.05). These results suggest that the PP4 phosphatase complex is ubiquitously expressed in rat tissues and likely involved in adaptation to or disease associated with high-altitude hypoxia.
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Objective:To investigate whether salidroside (SAL) improves lung tissue injury in rats with severe pneumonia (SP) through mediating toll-like receptor 4/nuclear transcription factor-κB/NOD-like receptor protein 3 (TLR4/NF-κB/NLRP3) signaling pathway.Methods:Seventy-five Wistar rats were used in this study. Fifteen of them were randomly selected as the sham operation group, and the others were induced by endotracheal infusion of Klebsiella pneumoniae ( Kp) suspension to construct a rat model of SP. After modeling, the rats were randomly divided into four groups with 15 rats in each group: model group, low-dose SAL group (30 mg/kg), high-dose SAL group (60 mg/kg) and dexamethasone (DXMS, 15 mg/kg) group. The sham operation group and the model group were given the same amount of normal saline for seven consecutive days. The wet-dry weight ratio (W/D) of lung tissues in each group was detected. HE and TUNEL staining methods were used to observe the morphology of lung tissues and cell apoptosis. The levels of TNF-α, IL-1β, IL-6, IL-18 and IL-10 in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The expression of TLR4, myeloid differentiation factor (MyD88), NF-κBp65, phosphorylated NF-κBp65 (p-NF-κBp65) and NLRP3 at protein level in lung tissues was detected by Western blot. Results:The rat model of SP was successfully constructed by endotracheal infusion of Kp suspension. Compared with the sham operation group, the model group showed more severe edema of lung tissues, increased W/D value ( P<0.05), loose and incomplete alveolar structure, edema of alveolar wall and thickened alveolar wall, massive inflammatory cell infiltration, increased apoptosis rate as well as higher levels of TNF-α, IL-1β, IL-6 and IL-18 and lower lover of IL-10 in BALF. Moreover, the relative expression of TLR4, MyD88, NF-κBp65, p-NF-κBp65 and NLRP3 at protein level in lung tissues was increased in the model group ( P<0.05). Gradually improved pathological injury of lung tissues, decreased W/D value ( P<0.05), recovered alveolar structure, reduced alveolar wall edema and decreased cell apoptosis rate were observed in the low-dose and high-dose SAL groups as well as the DXMS group as compared with those of the model group. Besides, the three groups also showed decreased levels of TNF-α, IL-1β, IL-6 and IL-18 and increased level of IL-10 in BALF, and inhibited expression of TLR4, MyD88, NF-κBp65, p-NF-κBp65 and NLRP3 at protein level in lung tissues ( P<0.05). DXMS performed better in improving lung injury in rats with SP, followed by high and low doses of SAL ( P<0.05). Conclusions:SAL could reduce cell apoptosis and improve the Kp-induced lung injury in rats. The mechanism might be related to the blockage of TLR4/NF-κB/NLRP3 signaling pathway activation and inhibition of inflammatory factor expression.
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Acute lung injury ( ALI) and its most extreme form a-cute respiratory distress syndrome ( ARDS) are lung diseases with high morbidity and mortality. There is no effective therapeutic intervention until now for its complicated pathophysiologi-cal processes and sophisticated regulatory mechanism. Histone deacetylases (HDACs) are a family of proteins with deacetylase activity. Studies have shown that HDACs are involved in the pathophysiological processes of ALI/ARDS, including inflammatory responses,endothelial permeability,oxidative stresses,alveolar fluid clearance and lung tissue repairment. Simultaneously, the use of HDACs inhibitors (HDACIs) can interfere with ALI/ ARDS progression. In this review we describe and summarize the pathophysiological processes and the underlying mechanisms in ALI/ARDS regulated by HDACs and HDACIs in detail, in order to provide the basis for the clinical application of HDACs-targe- ted agents and indicate directions for future study.
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@#Abstract: Objective To compare the efficiency of lung-microscopy and tissue homogenate in the detection of Angiostrongylus cantonensis larvae from Pomacea canaliculate, with the amin of finding a simple and rapid method suitable for different scenarios. Methods Pomacea canaliculata was caught and collected from ponds, ditches, rivers and other environments in the confirmed epidemic areas reported by predecessors. After each snail was weighed one by one, and dissected into two parts: lung sac and muscle. Firstly, each snail lung sac was check for nodules with lung-microscopy, and the nodules were picked out with anatomical needle and press them separately. Then, the worm was found and identified for worm species under microscope, and the lung sac and snail meat then was rechecked with tissue homogenate. Results A total of 330 snails were detected, with 19.1% (63/330) snails with Angiostrongylus cantonensis stage Ⅲ larvae were detected by tissue homogenate and 15.8% (52/330) snails with nodules were detected by lung-microscopy. Among them, 36 snails with nodules and larvae were detected by the lung-microscopy, and all of them were positive by the tissue homogenate, with a coincidence rate of 100% (36/36); 16 snails with nodules but no larvae, among which 6 snails were positive and 10 snails were negative by the tissue homogenate, The false detection rate was 19.2% (10/52). 278 snails with no nodules, but 21 of them were detected by the tissue homogenate, the missing rate was 7.6% (21/278). There was no significant difference between tissue homogenate and lung-microscopy (taking the positive determination of nodules as the standard) (χ2=1.27, P=0.26, P>0.05). There was significant difference between tissue homogenate and lung-microscopy (taking the detection of larvae as the standard)(χ2=8.66, P = 0.003, P<0.01). There was no significant difference between the two methods and tissue homogenate in the detection rate of large snails ( ≥25 g, χ2=0.08，P=0.777; χ2=2.58, P=0.108), but there was significant difference between the two methods and tissue homogenate in the detection rate of small snail (≤10 g, χ2=5.63, P=0.02). Conclusions Compared with the tissue homogenate, lung-microscopy is simple in the detection of large snails, requires less instruments and equipment, and its detection speed is faster. It is suitable for the field investigation of the natural focus of Angiostrongylus cantonensis. The tissue homogenate has high sensitivity, can directly display the insect state and activity, has strong insect vitality and high detection rate, it's more suitable for food safety risk monitoring.
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OBJECTIVE: To explore the therapeutic effect of tetrandrine(TET) on silicosis model rats and its toxic effect on liver and kidney function. METHODS: The specific pathogen free healthy male Wistar rats were randomly divided into the control group, the model group and the TET group, with 14 rats in each group. By un-exposure tracheal injection method, the rats in the model and TET groups were given one-time tracheal infusion of free silicon dioxide suspension with a mass concentration of 50 g/L to establish the rat model of silicosis. Rats in the control group were infused with 1 mL of 0.9% sodium chloride solution with the same method. On the second day after the model was established, the TET group was given 30 mg/kg body mass of TET solution by gavage. The other two groups were given the same amount of 0.9% sodium chloride solution. The treatment was once per day, six times per week. Seven rats in each group were sacrificed on the 28 th and 56 th days after modeling. The morphological change of the lung, liver and kidney tissues of each group was observed. The enzyme-linked immunosorbent assay was used to detect the level of tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1), interleukin(IL)-1β and IL-6, in the lung tissues of rats in each group. The activities of aminotransferase(ALT), aspartate aminotransferase(AST) and the levels blood urea nitrogen(BUN), creatinine(CRE) were detected by automatic biochemical analyzer. RESULTS: The lung organ coefficients of rats in the TET group were lower than those of the model group on the 28 th and 56 th days(all P<0.05). The lung organ coefficient of the rats in the TET group on the 56 th day was higher than that in the same group on the 28 th day(P<0.05). The lung tissue structure of the control group was normal. After modeling, the lung tissues of rats in model group showed different degrees of pathological changes such as alveolar structure destruction, inflammatory cell infiltration, and fibrosis on the 28 th and 56 th days. The degree of pathological changes in TET group was less than that of the model group. In the lung tissues of rats in the model group, the levels of TNF-α, TGF-β1, IL-1β and IL-6 were higher than those of the control group(all P<0.01). The levels of TNF-α, TGF-β1, IL-1β and IL-6 in the lung tissues of rats in the TET group were lower than that of the model group(all P<0.01), but there was no statistically significant difference when compared with the control group(all P>0.05). The activities of ALT and AST in the TET group were higher than those in the model group and the control group(all P<0.01). The level of serum BUN in TET group was higher than that in control group(P<0.01), but it showed no statistical difference when compared with the control group(P>0.05). The level of serum CRE in each group showed no significant difference(P>0.05). There were no abnormal pathological changes found in the liver and kidney tissues of rats in each group at different times. CONCLUSION: TET can reduce the inflammatory response in silicosis rats and improve lung tissue fibrosis; however, the therapeutic dose may have certain toxicity to the liver and kidney of the silicosis rats.
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To study the effect of mineral Chloriti Lapis on pulmonary metabolites and metabolic pathways in lung tissues of rats with acute exacerbation of chronic obstructive pulmonary disease(AECOPD). The AECOPD rat model of phlegm heat syndrome was replicated by the method of smoking combined with Klebsiella pneumoniae infection. Except for using UPLC-Q-TOF-MS analysis, SPSS 18.0, SIMCA 13.0 and other software were also used for statistical analysis. Through literature search and online database comparison, the differential metabolites were identified, and the possible metabolic pathways were analyzed. After 15 days of administration, PLS-DA analysis was carried out on lung tissue samples of rats in each group. The results showed that the metabolic profiles of lung tissues of rats in each group could be well separated, which indicated that Chloriti Lapis and aminophylline had significant intervention effect on the lung metabolic profile of rats with AECOPD. Moreover, the metabolic profile of Chloriti Lapis group was closer to that of control group, and the intervention effect was better than that of aminophylline group. As a result, 15 potential differential metabolites were identified: phytosphingosine, sphinganine, tetradecanoylcarnitine, L-palmitoylcarnitine, elaidic carnitine, lysoPC[18∶2(9Z,12Z)], lysoPC(16∶0), lysoPC[18∶1(9Z)], lysoPC(18∶0), stearic acid, lysoPC(15∶0), arachidonic acid, docosapentaenoic acid, linoleic acid and palmitic acid. Among them, Chloriti Lapis could significantly improve the levels of 10 differential metabolites of phytosphingosine, tetradecanoylcarnitine, L-palmitoylcarnitine, elaidic carnitine, lysoPC[18∶2(9Z,12Z)], lysoPC(16∶0), lysoPC[18∶1(9Z)], stearic acid, lysoPC(15∶0), and palmitic acid(P<0.05). The intervention effect of Chloriti Lapis group was better than that of aminophylline group. Analysis of metabolic pathways showed that there were 8 possible metabolic pathways that could be affected, and three of the most important metabolic pathways(pathway impact>0.1) were involved: linoleic acid metabolism, arachidonic acid metabolism, and sphingolipid metabolism. Chloriti Lapis had obvious intervention effects on lung tissue-related metabolites and metabolic pathways in rats with AECOPD, and the effect was better than that of aminophyllinne.
الموضوعات
Animals , Rats , Lung , Medicine, Chinese Traditional , Metabolomics , Minerals , Pulmonary Disease, Chronic Obstructiveالملخص
Objective:To investigate the expression changes at the transcriptional level in normal lung tissues of mice after exposure to heavy ion radiation for different durations at different doses, aiming to provide evidence for exploring sensitive genes of heavy ion radiation, heavy ion radiation effect and the damage mechanism.Methods:Experiments on the temporal kinetics: the whole thorax of mice was irradiated with 14.5Gy carbon-ions and the total RNA of lung tissue was extracted at 3days, 7days, 3 weeks and 24 weeks. In dose-dependent experiment, the total RNA of lung tissue was extracted at 1 week after irradiated with a growing thoracic dose of 0, 7.5, 10.5, 12.5, 14.5, 17.5 and 20Gy. Protein-to-protein interaction (PPI) analysis and gene-ontology biological process enrichment analysis were performed on significant differentially expressed genes (DEGs).Results:A clearly differential expression patterns were observed at 3-day (acute stage), 1-week (subacute stage), 3-week (inflammatory stage) and 24-week (fibrosis stage) following 14.5Gy carbon-ions irradiation. Among those, the 3-day time point was found to be the mostly different from the other time points, whereas the 7-day time point had the highest uniformity with the other time points. Cellular apoptosis was the main type of cell death in normal lung tissues following carbon-ions exposure. The interactive genes of Phlda3, GDF15, Mgmt and Bax were identified as the radiosensitive genes, and Phlda3 was the center ( R=0.76, P<0.001). Conclusion:The findings in this study provide transcriptional insights into the biological mechanism underlying normal lung tissue toxicity induced by carbon-ions.
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OBJECTIVE: To establish an LC-MS/MS method for the determination of arformoterol tartrate in rat lungs and study the pharmacokinetics of arformoterol tartrate in rat lungs. METHODS: Lung samples were collected at different time points after SD rats were given nebulized infusion solution of arformoterol tartrate. An LC-MS/MS method was established and validated to analyze the drug concentrations,and pharmacokinetic parameters were calculated by professional software named DAS3.0 pharmacokinetic program. RESULTS: The main pharmacokinetic parameters of arformoterol tartrate in the lungs of rats were as follows: t1/2 was 15.55 h, cmax was 16.48 ng•g-1, AUC0-t was 48.41 ng•h•g-1, and MRT0-t was 5.45 h. CONCLUSION: After aerosolization of arformoterol tartrate inhalation solution, the drugis rapidly distributed in the lung, has a long half-life and can be maintained for a certain period of time, suggesting that atomized administration of arformoterol tartrate can be used for the treatment of chronic obstructive pulmonary disease.
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RESUMO Objetivo O objetivo deste estudo foi investigar os efeitos agudos e crônicos da vareniclina no tecido pulmonar em um estudo experimental. Métodos Um total de 34 ratos foi alocado aleatoriamente em grupos de estudo (vareniclina) e controle. Assim, os ratos foram divididos em dois grupos: (i) grupo controle e (ii) grupo vareniclina. A seguir, os ratos de cada grupo foram, por sua vez, subdivididos igualmente em agudos (C1; V1) e crônicos (C2; V2), e todos os ratos dos grupos agudos e crônicos foram sacrificados sob anestesia: no 45.º dia, para o grupo agudo [C1 (n=5) e V1 (n=12)], e no 90.º dia, para o grupo crônico [C2 (n=5) e V2 (n=12)], respectivamente. Em seguida, foram realizadas análises bioquímicas e histopatológicas. Resultados Trinta e quatro ratos completaram o estudo. Destes ratos, 24 estavam no grupo vareniclina e 10 no grupo controle. Na exposição crônica à vareniclina, os níveis de oxidante composto por malondialdeído (MDA) e mieloperoxidase (MPO) aumentaram, e os níveis de superóxido dismutase (SOD), catalase (CAT), glutationa (GSH) e glutationa peroxidase (GPx), nomeados como antioxidantes, diminuiram significativamente quando comparados com o grupo controle. Os níveis de MDA e MPO também foram significativamente mais elevados e os níveis de SOD, CAT, GPx e GSH foram significativamente mais baixos no grupo vareniclina crônico, quando comparado ao grupo vareniclina agudo. Estes achados também foram confirmados por observações histopatológicas. Conclusões Este é o primeiro estudo que avaliou os efeitos pulmonares da vareniclina experimentalmente em um modelo animal. Observamos que o tratamento crônico da vareniclina causa inflamação e lesão pulmonar.
ABSTRACT Objective This study aimed to investigate acute and chronic effects of varenicline on lung tissue in an experimental study. Methods A total of 34 rats were randomly allocated into study (varenicline) and control groups. The rats were divided into two groups (i) control group, (ii) varenicline group. Then, the rats in the each group were sub-divided equally in turn as acute (C1; V1) and chronic (C2; V2) ; all rats of acute and chronic groups were sacrificed under the anesthesia on the 45th day for acute group [C1 (n=5) and V1 (n=12)] and the 90th day for chronic group [C2 (n=5) and V2 (n=12)], respectively. Thus, biochemical and histopathological analysis were carried out. Results Thirty four rats completed the study, 24 were in varenicline group and 10 were in control group. In chronic exposure to varenicline, oxidant levels comprising of malondialdehyde (MDA), and myeloperoxidase (MPO) increased and superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and glutathione peroxidase (GPx) levels, named as antioxidants, decreased significantly when compared to the control group. MDA and MPO levels were also significantly higher and SOD, CAT, GPx, GSH levels were also significantly lower in chronic varenicline group when compared to acute varenicline group. These findings were also supported by histopathological observations. Conclusion This is the first study, which evaluated pulmonary effects of varenicline experimentally on an animal model. It was observed that chronic varenicline treatments cause inflammation and lung cell injury.
الموضوعات
Animals , Rats , Superoxide Dismutase/blood , Varenicline/pharmacology , Lung/drug effects , Catalase/blood , Oxidative Stress , Glutathione , Glutathione Peroxidase , Malondialdehyde/bloodالملخص
This study investigates the effect of the overexpression of the placental growth factor (PGF) and hyperoxia on lung development and determines whether anti-PGF antibody ameliorates hyperoxia-mediated impairment of lung development in newborn rats. After exposure to normoxic conditions for seven days, newborn rats subjected to normoxia were intraperitoneally or intratracheally injected with physiological saline, adenovirus-negative control (Ad-NC), or adenovirus-PGF (Ad-PGF) to create the Normoxia, Normoxia+Ad-NC, and Normoxia+Ad-PGF groups, respectively. Newborn rats subjected to hyperoxia were intraperitoneally injected with physiological saline or anti-PGF antibodies to create the Hyperoxia and Hyperoxia+anti-PGF groups, respectively. Our results revealed significant augmentation in the levels of PGF and its receptor Flt-1 in the lung tissues of newborn rats belonging to the Normoxia+Ad-PGF or Hyperoxia groups. PGF overexpression in these groups caused lung injury in newborn rats, while anti-PGF antibody treatment significantly cured the hyperoxia-induced lung injury. Moreover, PGF overexpression significantly increased TNF-α and Il-6 levels in the bronchoalveolar lavage (BAL) fluid of the Normoxia+Ad-PGF and Hyperoxia groups. However, their levels were significantly reduced in the BAL fluid of the Hyperoxia+anti-PGF group. Immunohistochemical analysis revealed that PGF overexpression and hyperoxia treatment significantly increased the expression of the angiogenesis marker, CD34. However, its expression was significantly decreased upon administration of anti-PGF antibodies (compared to the control group under hyperoxia). In conclusion, PGF overexpression impairs lung development in newborn rats while its inhibition using an anti-PGF antibody ameliorates the same. These results provided new insights for the clinical management of bronchopulmonary dysplasia in premature infants.
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Animals , Female , Pregnancy , Rats , Autoantibodies/metabolism , Hyperoxia/metabolism , Lung Injury/metabolism , Placenta Growth Factor/metabolism , Antibodies, Monoclonal/metabolism , Autoantibodies/immunology , Microscopy, Electron, Scanning , Hyperoxia/complications , Hyperoxia/diagnostic imaging , Disease Models, Animal , Lung Injury/pathology , Lung Injury/diagnostic imaging , Placenta Growth Factor/immunology , Animals, Newborn , Antibodies, Monoclonal/immunologyالملخص
SUMMARY OBJECTIVES This study was conducted to reveal the possible protective effects of ticagrelor and enoxaparin pretreatment against ischemia-reperfusion (IR)-induced injury on the lung tissue of a rat model. METHODS Wistar albino rats were randomly divided into 4 groups as follows: group-1 (control-sham), group-2 (control-saline+IR), group-3 (ticagrelor+IR), group-4 (enoxaparin+IR). Before the ischemic period, saline, ticagrelor, and enoxaparin were administered to the 2nd-4th groups, respectively. In these groups, IR injury was induced by clamping the aorta infrarenally for 2 h, followed by 4 h of reperfusion except group-1. After the rats were euthanized, the lungs were processed for histological examinations. Paraffin sections were stained with Haematoxylin&Eosin (H&E) for light microscopic observation. Apoptosis was evaluated by caspase-3 immunoreactivity. Data were statistically analyzed using the SPSS software. RESULTS In the lung sections stained with H&E, a normal histological structure was observed in group-1, whereas disorganized epithelial cells, hemorrhage, and inflammatory cell infiltration were seen in the alveolar wall in group-2. The histologic structure of the treatment groups was better than that of group-2. Caspase-3(+) apoptotic cells were noticeable in sections of group-2 and were lower in the treatment groups. In group-4, caspase-3 immunostaining was lower than in group-3. In group-2, apoptotic cells were significantly higher than in the other groups (p<0.001). CONCLUSION Based on the histological results, we suggested that both therapies ameliorated the detrimental effects of IR. Caspase-3 immunohistochemistry results also revealed that pre-treatment with enoxaparin gave better results in an IR-induced rat injury model. In further studies, other parameters such as ROS and inflammatory gene expressions should be evaluated for accurate results.
RESUMO OBJETIVOS Este estudo foi realizado para revelar os possíveis efeitos protetores do ticagrelor e do pré-tratamento da enoxaparina no tecido pulmonar contra o modelo de lesão induzida por isquemia-reperfusão (IR). MÉTODOS Ratos albinos Wistar foram randomizados e divididos em quatro grupos: grupo 1 (controle-sham), grupo 2 (controle-salina + IR), grupo 3 (ticagrelor + IR), grupo 4 (enoxaparina + IR). Antes do período isquêmico, salina, ticagrelor e enoxaparina foram administrados nos grupos 2-4, respectivamente. Nesses grupos, a lesão de IR foi induzida pelo clampeamento da aorta na região da infrarrenal por duas horas, seguida por quatro horas de reperfusão, exceto no grupo 1. Após a sacrificação, os pulmões foram processados para exames histológicos. Secções de parafina foram coradas com hematoxilina e eosina (H&E) para observação microscópica de luz. A apoptose foi avaliada pela imunorreatividade da caspase-3. Os dados foram analisados estatisticamente pelo programa SPSS. RESULTADOS Nas secções pulmonares coradas com H&E, estrutura histológica normal foi observada no grupo 1, enquanto células epiteliais desorganizadas, hemorragia e infiltração de células inflamatórias foram observadas na parede alveolar no grupo 2. A estrutura histológica dos grupos de tratamento foi melhor que o grupo 2. Células apoptóticas caspase-3 (+) foram notadas em secções do grupo 2, e essas células foram mais baixas nos grupos de tratamento. No grupo 4, a imunocoloração com caspase-3 foi menor que no grupo 3. No grupo 2, as células apoptóticas foram significativamente maiores que nos outros grupos (p<0,001). CONCLUSÃO Com base nos resultados histológicos, sugerimos que ambas as terapias atenuaram os efeitos prejudiciais da RI. Resultados de imuno-histoquímica com caspase-3 também revelaram que o pré-tratamento com enoxaparina proporcionou melhores resultados no modelo de lesão induzida por IR. Em estudos posteriores, outros parâmetros, como ROS e expressões gênicas inflamatórias, devem ser avaliados quanto a resultados precisos.
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Animals , Male , Aorta, Abdominal/surgery , Reperfusion Injury/prevention & control , Enoxaparin/pharmacology , Protective Agents/pharmacology , Ticagrelor/pharmacology , Lung/drug effects , Reperfusion Injury/pathology , Random Allocation , Rats, Wistar , Apoptosis/drug effects , Disease Models, Animal , Caspase 3/metabolism , Lung Injury/prevention & control , Lung/pathologyالملخص
OBJECTIVE@#To investigate the effect of ulinastatin on the inflammatory mediators and their signaling pathways miR-146a/TLR4/NF-κB in rats with hemorrhagic shock.@*METHODS@#Seventy-two SD rats were randomly assigned into shock without resuscitation group (SR group, =24), acetated Ringer's solution resuscitation group (AR group, =24) and ulinastatin treatment group (=24). In all the 3 groups hemorrhagic shock models were established by femoral artery bleeding (with the mean arterial pressure maintained at 30-40 mmHg) without resuscitation (in SR group) or with resuscitation (in AR and ulinastatin groups) using acetated Ringer's solution for 30 min at 60 min after the onset of shock. At 1, 4, and 6 h after the shock onset or immediately after shock if the rats died, the lung tissues were taken for measurement of mRNA expressions of miR-146a, tumor necrosis factor- (TNF-), interleukin-1 (IL-1), IL-4, IL-6 and IL-10 using real-time quantitative PCR and the protein expressions of TLR4, MyD88, IκB-, p-IκB-, NF-κB p65, IRAK4, p-IRAK4 (Thr345, Ser346), p-IRAK4 (Thr342) and TRAF6 using Western blotting. The lung histopathology of the rats was examined under optical microscope with HE staining.@*RESULTS@#Compared with the SR group, the rats in the AR group showed slightly alleviated inflammatory infiltration in the lung tissues with significantly increased mRNA levels of miR-146a, IL-4 and IL-10 ( < 0.05) and protein expressions of IκB-, p-IRAK4 (Thr342) and p-IRAK4 (Thr345, ser346) ( < 0.05), and decreased mRNA levels of TNF-, IL-1 and IL-6 ( < 0.05) and protein expressions of TLR4, MyD88, NF-κB p65, p-IκB-, IRAK-4 and TRAF6 ( < 0.05). Compared with those in AR group, the rats in ulinastatin group showed further alleviation of inflammatory lung tissue injury, with increased mRNA levels of miR-146a, IL-4 and IL-10 ( < 0.01) and protein expressions of IκB-, p-IRAK4 and p-IRAK4 ( < 0.01) and decreased mRNA levels of TNF-, IL-1 and IL-6 ( < 0.01) and protein expressions of TLR4, MyD88, NF-κB p65, p-IκB-, IRAK-4 and TRAF6 ( < 0.01).@*CONCLUSIONS@#Ulinastatin combined with acetated Ringer's solution resuscitation alleviates lung inflammations in rats with hemorrhagic shock possibly by enhancing miR-146a expression to regulate TLR4/NF-κB signaling pathway through a negative feedback mechanism and thus modulate the balance of pro-inflammatory and anti-inflammatory factors.
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Bronchopulmonary dysplasia (BPD) is the most common chronic respiratory disease in premature and low birth weight infants,characterized by alveolar and pulmonary vascular dysplasia.This article reviews the factors related to the pathogenesis of BPD,with the aim of providing new ideas for the research of pathogenesis of BPD and its prevention and treatment.Among them,immature lung development,acute lung injury,and abnormal repair after injury are the key links of BPD.Other influencing factors include oxygen poisoning,barotrauma,infection,lack of nutritional support,patent ductus arteriosus,blood transfusion,gastroesophageal reflux,pulmonary interstitial edema,abnormal coagulation function,cholestasis and so on.
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Bronchopulmonary dysplasia(BPD)is the most common chronic respiratory disease in premature and low birth weight infants, characterized by alveolar and pulmonary vascular dysplasia.This article reviews the factors related to the pathogenesis of BPD, with the aim of providing new ideas for the research of pathogenesis of BPD and its prevention and treatment.Among them, immature lung development, acute lung injury, and abnormal repair after injury are the key links of BPD.Other influencing factors include oxygen poisoning, barotrauma, infection, lack of nutritional support, patent ductus arteriosus, blood transfusion, gastroesophageal reflux, pulmonary interstitial edema, abnormal coagulation function, cholestasis and so on.
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Objective: To qualitative analze the chemical components in rat lung tissues after oral administration Qinbai Qingfei Concentrated Pellets (QQCP). Methods: The rat lung tissues were collected and analyzed by UPLC-Q-TOF-MS after ig administration of QQCP. The chemical components in control and dose groups were identified and speculated by Peakview and Metabolite Pilot data processing software using retention time, exact relative molecular mass, and cleavage fragments of MS/MS as indexes. Results: A total of 25 compounds were identified, including 17 prototypes and eight metabolites. Conclusion: The chemical constituents in the rat lung tissues were identified after oral administration of QQCP based on UPLC-Q-TOF-MS, which will contribute to elucidate the effects of potential pharmacodynamic material basis of QQCP on mycoplasma pneumoniae.
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AIM:To observe the effects of ginsenoside Rh 1 on the levels of inflammatory factors in serum and bronchoalveolar lavage fluid(BALF),and the pathological changes of the lung tissues in an experimentally induced mouse asthma model.METHODS:Male BALB/c mice(n=40)were divided into 4 groups:normal control group,asthma mo-del group,and low-dose(40 mg· kg-1 · d-1 )and high-dose(80 mg· kg-1 · d-1 )ginsenoside Rh1 groups.The bron-chial asthma mouse model was established by the method of ovalbumin induction and excitation ,and during the excitation period,the mice were daily treated with ginsenoside Rh 1 for 2 weeks.At 24 h after the final dose of ginsenoside Rh 1,the mice were sacrificed.The number of eosinophils(EOS)and the concentrations of interleukin(IL)-4,IL-5 and interferon(IFN)-γin BALF were determined.The levels of IgG and IgE in serum were measured ,and the expression of transforming growth factor(TGF)-β1 and the pathological changes in lung tissues were evaluated.RESULTS:Ginsenoside Rh1 inhibi-ted the increases in the number of EOS and the concentrations of IL-4,IL-5,IFN-γand IgE,reversed the increased ex-pression of TGF-β1,and improved the pathological changes of the lung tissues in asthmatic mice.CONCLUSION:Gin-senoside Rh1 improves the immuno-inflammatory profile and pathological changes in the experimentally induced mouse asth -ma model,implying its potential therapeutic effect on asthma.
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Objective To study the effects of acetated ringer's solution resuscitation in hemorrhagic shock rats on inflammatory mediators on lung tissue and their JNK (c-Jun N-terminal kinase) signaling pathways. Methods Thirty-two SD rats were randomly(random number) divided into four groups: shock without resuscitation group (CR, n=8), saline group (NR, n=8), lactated ringer's solution group (LR, n=8) and acetated ringer's solution group (AR,n=8). The rats of NR group, LR group and AR group were prepared into shock models (mean arterial blood pressure maintained at 40-45 mmHg),The rats of NR group, LR group and AR group were in the shock for 60 min and then the corresponding kinds of liquid were administered for 30 min and observation was carried out for 4 hours. The rats of CR group without liquid resuscitation were observed for 4 hours after shock. After that, the lung tissues of rats were taken from NR group, LR group and AR group as well as from CR group 4 hours after shock (if the rats died, the lung tissues were immediately taken). The levels of TNF-α, IL-4 and IL-10 mRNA in lung were measured by real-time polymerase chain reaction (RT-PCR),and Western blot was used to measure the levels of JNK phosphorylation and MKP-1 acetylation. The one-way ANOVA was used for comparison among groups. Between the two groups, the comparison was analyzed by using LSD-t test. Results The IL-4 mRNA expression of lung tissue in AR group was higher than that in CR group, NR group and LR group (CR group:0.42±0.34; NR group:2.60±0.66; LR group:6.24±2.95; AR group: 11.08±4.24; P＜0.05).The IL-10 mRNA expression of lung tissue in AR group was significantly higher than that in CR group, NR group and LR group (CR group:0.25±0.25; NR group:2.79±1.62; LR group:3.51±1.66; AR group:9.35±2.86;P＜0.01).The TNF-a mRNA expression in AR group was significantly lower than that in CR group, NR group and LR group (CR group:4.98±1.26; NR group:2.50±0.76; LR group:3.87±3.00; AR group:0.19±0.09; P＜0.01). The level of JNK phosphorylation in lung tissue of rats in AR group was significantly lower than that in CR group, NR group and LR group (CR group:0.52±0.12; NR group:0.42±0.08; LR group:0.30±0.08; AR group:0.17±0.06;P＜0.01). The level of MKP-1 acetylation in lung tissue of rats in AR group was significantly higher than that in CR group, NR group and LR group (CR group:0.14±0.07; NR group:0.30±0.07; LR group:0.37±0.02; AR group:0.48±0.06;P＜0.01). Compared with normal saline and lactated ringer's solution, acetated ringer's solution used in hemorrhagic shock rats could promote MKP-1 acetylation, inhibit the phosphorylation of JNK, significantly inhibit the lung tissue TNF-a released, promote the release of anti-inflammatory factors, IL-4 and IL-10. Conclusions The acetated ringer's solution for resuscitation of hemorrhagic shock in rats could reduce inflammation of lung tissue in a certain extent, probably by enhanced the acetylation of MKP-1 to inhibited JNK signaling pathway and reduced lung tissue inflammation.
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Objective To investigate the effect of isosorbide mononitrate on the levels of NO,iNOS,IL-1 and IL-6 in lung tissue of spontaneously hypertensive rats (SHR).Methods Fourteen-week-old Wistar and SHR male rats were randomly divided into the W0,W1,S0 and S1 group,with 10 rats ineach group.Rats in the W0 and S0 group were fed with the normal saline and the ordinary food,rats in the W1 and S1 group were fed with isosorbide mononitrate and the ordinary food.Twelve weeks later,levels of NO,iNOS,IL-1 and IL6 in rat lung tissue were detected.Results Compared with the W0 group,levels of NO,iNOS,IL-1 and IL6 were significantly increased in the W1 groups (P ＜ 0.05,respectively).Compared with the SO group,levels of NO,iNOS,IL-1 and IL6 were significantly increased in the S1 group (P ＜ 0.05,respectively).In the W1 and S1 group,levels of iNOS and NO were positively correlated with IL-1 and IL-6.Conclusion 1.Isosorbide mononitrate may lead to increases of NO,iNOS,IL-1 and IL6 in lung tissue of Wistar rats,which indicates the presence of chronic inflammation.2.Longterm feeding of isosorbide mononitrate may lead to increases of inflammatory factors in SHR rats,contributing to the inflammatory state in rats.
الملخص
Objective To investigate the effects of chronic hypobaric hypoxia on COX1 protein and oxidative stress in lung tissue. Methods Rats were randomly divided into normoxia group (1 500 m)and hypoxia group (4 300 m). The rats in hypoxia group were sampled after exposure to hypoxia for 30 days. The COX1 protein in lung tissue of rats was determined by Western blot method; HIF-1αlevel in lung tissue and serum, ROS in plas-ma and SOD, CAT enzyme in lung tissue were determined by ELISA method; PAP of rats were determined by physiological signal acquisition system. Results HIF-1 protein express in serum and lung tissue of rats in hypoxia group was significantly higher than that of the normoxia group (P<0.01), the content of ROS was significantly lower than that of the normoxia group (P<0.01), the expression of COX1 protein in lung tissues of hypoxia group was significantly decreased(P<0.01),serum total antioxidant capacity was elevated in hypoxia group(P<0.01).Conclusions The effects of chronic hypobaric hypoxia on lung tissue may be caused by direct injury, not only by oxidative stress.