Bronchoalveolar Lavage Fluid Of Rats Research Articles

Protective Effect of Colchicine on LPS-Induced Lung Injury in Rats via Inhibition of P-38, ERK1/2, and JNK Activation

Introduction: Acute lung injury (ALI), a commonly detected syndrome, is characterized by the accumulation of neutrophils and leucocytes, and inflammation of pulmonary tissues. Objective: The present study was designed to investigate the effect and underlying mechanism of colchicine on LPS-induced lung injury. Methods: The rats were divided randomly into 6 groups of 10 each: normal control, untreated, and 4 colchicine (5, 10, 15, and 20 mg/kg) treatment groups. ALI was induced in rats by the administration of 20 μg LPS intratracheally. Rats in the normal control and untreated groups were injected normal saline, while those in the treatment groups received 5, 10, 15, and 20 mg/kg doses of colchicine daily for 1 month. ELISA was used for determination of interleukin (IL)-1β, IL-6, tumour necrosis factor (TNF)-α, superoxide dismutase (SOD), and leucocytes in the rat bronchoalveolar lavage fluid (BALF). The expression of P-38, JNK, and Erk-1/2 was analysed by Western blotting. Results: In LPS-administered TC-1 cells, the levels of IL-1β, IL-6, and TNF-α were markedly higher. Treatment with colchicine reduced the levels of IL-1β, IL-6, and TNF-α in LPS-administered TC-1 cells. Colchicine treatment caused a marked reduction in LPS-induced accumulation of inflammatory cells in the rat lungs. The LPS-induced aggregation of leucocytes and neutrophils in the rat BALF was also suppressed markedly on treatment with colchicine. Treatment of the lung injury in rats with colchicine caused a marked decrease in the level of IL-1β, IL-6, and TNF-α in BALF. The LPS-mediated suppression of SOD in the rat BALF was prevented by treatment with colchicine. Treatment of the rats with colchicine attenuated the LPS-induced activation of P-38, Erk1/2, and JNK in pulmonary tissues. Conclusion: In summary, colchicine treatment prevents LPS-induced lung damage in rats through targeting activation of P-38, ERK1/2, and JNK. Therefore, colchicine may be used for the development of treatment for ALI.

Chlorobenzoxime inhibits respiratory syncytial virus infection in neonatal rats via up-regulation of IFN-γ in dendritic cells

Purpose: To investigate the effect of chlorobenzoxime on respiratory syncytial virus (RSV) infection in vitro in lung alveolar cells and in vivo in neonatal rats, as well as the mechanism of action involved.
 Methods: RSV infection in neonatal rats was induced via intranasal administration of 2 x 106PFU viral particles. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting were used for determination of changes in interleukin expression.
 Results: RSV infection in BEAS-2B cells caused significant reduction in viability and marked alteration in morphological appearance (p < 0.05). Exposure of RSV-infected BEAS-2B cells to chlorobenzoxime prevented viability reduction and changes in morphology, and led to reductions in RSV-mediated increases in levels of interleukin-6 and interleukin-8. Moreover, RSV infection significantly enhanced ROS levels in BEAS-2B cells, when compared to control cells (p < 0.05). Chlorobenzoxime at a concentration of 30 μM completely suppressed RSV-mediated generation of ROS in BEAS-2B cells. In neonatal rats, RSV-induced upregulation of interleukin-4, interleukin-13 and TNF-α, were suppressed in bronchoalveolar lavage fluid (BALF) and lung tissues by chlorobenzoxime. Moreover, the RSVmediated reduction in IFN-γ was maximally blocked by chlorobenzoxime at a dose of 10 mg/mL. Chlorobenzoxime enhanced the proportion of IFN-γ -producing cells in neonatal rat BALF.
 Conclusion: Chlorobenzoxime exhibits antiviral against RSV infection in neonatal rats via increase in dendritic cell population, leading to inhibition of cytokine production. Therefore, chlorobenzoxime is a potential therapeutic agent for RSV infection.
 Keywords: Respiratory syncytial virus, Cytokines, Dendritic cells, Lung aveolar cells, Morphology, Interleukins

Resveratrol protects against asthma-induced airway inflammation and remodeling by inhibiting the HMGB1/TLR4/NF-κB pathway.

The aim of the present study was to explore the protective role of resveratrol (RES) in asthma-induced airway inflammation and remodeling, as well as its underlying mechanism. An asthma rat model was induced by ovalbumin (OVA) treatment. Rats were randomly assigned into sham, asthma, 10 µmol/l RES and 50 µmol/l RES groups. The amount of inflammatory cells in rat bronchoalveolar lavage fluid (BALF) was detected. Pathological lesions in lung tissues were accessed by hematoxylin and eosin (H&E), and Masson's trichrome staining. Levels of inflammatory factors in lung homogenate were detected via ELISA. The blood serum of asthmatic and healthy children was also collected for analysis. Reverse transcription-quantitative polymerase chain reaction was performed to detect high mobility group box 1 (HMGB1), Τoll-like receptor 4 (TLR4), myeloid differentiation primary response gene 88 (MyD88) and NF-κB expression in asthmatic and healthy children, as well as rats of the different groups. H&E staining demonstrated that multiple inflammatory cell infiltration into the rat airway epithelium of the asthma group occurred whilst the 50 µmol/l RES group displayed alleviated pathological lesions. Masson staining indicated that there was an increased airway collagen deposition area in the asthma and 10 µmol/l RES groups compared with the 50 µmol/l RES group. The number of inflammatory cells in BALF extracted from rats of the asthma and 10 µmol/l RES groups was higher compared with the 50 µmol/l RES group. Treatment with 50 µmol/l RES significantly decreased the thicknesses of the airway wall and smooth muscle. ELISA results illustrated that interleukin (IL)-1, IL-10 and tumor necrosis factor-α (TNF-α) levels were elevated, whereas IL-12 level was reduced in lung tissues of the asthma and 10 µmol/l RES groups whilst the 50 µmol/l RES group demonstrated the opposite trend. HMGB1, TLR4, MyD88 and NF-κB mRNA levels were remarkably elevated in rats of the asthma and 10 µmol/l RES groups compared with the 50 µmol/l RES group. Serum levels of IL-1, IL-10 and TNF-α were elevated, whereas IL-12 was reduced in asthmatic children compared with healthy children. The present results demonstrated that a large dose of RES alleviated asthma-induced airway inflammation and airway remodeling by inhibiting the release of inflammatory cytokines via the HMGB1/TLR4/NF-κB pathway.

Changes in TNF-α, IL-6, IL-10 and VEGF in rats with ARDS and the effects of dexamethasone.

Changes in tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and vascular endothelial growth factor (VEGF) in serum and bronchoalveolar lavage fluid (BALF) in rats with acute respiratory distress syndrome (ARDS) and the intervention effect of dexamethasone were observed to explore the theoretical basis of dexamethasone in the treatment of ARDS. Seventy-two rats were randomly divided into normal control group (group N, n=24), ARDS model group (group L, n=24) and dexamethasone group (group D, n=24). The ARDS rat model was established by jointly injecting oleic acid and lipopolysaccharide via the caudal vein, while rats in group D received intervention with dexamethasone. The wet/dry weight ratios of lung tissues were measured, and the levels of TNF-α, IL-6, IL-10 and VEGF in serum and BALF were measured via enzyme-linked immunosorbent assay. The wet/dry weight ratio of lung tissues of rats in group D was significantly decreased compared with that in group L (P<0.05 or P<0.01). The levels of TNF-α, IL-6 and VEGF in serum and BALF of rats in group L and D were obviously increased compared with those in group N at each time point (P<0.01). The levels of TNF-α, IL-6 and VEGF in serum and BALF of rats in group D were significantly decreased compared with those in group L (P<0.01). In conclusion, there is a serious imbalance between anti-inflammatory response and inflammatory response in rats with ARDS induced by oleic acid combined with lipopolysaccharide of Escherichia coli, whereas dexamethasone can alleviate lung injury through inhibiting expression levels of inflammatory factors and promoting expression levels of anti-inflammatory factors.

Chitosan oligosaccharides alleviate PM2.5-induced lung inflammation in rats.

Air pollution of particulate matter (PM), especially PM2.5, has become a major public health problem in China. Exploration of therapeutic and preventive measures against PM2.5 toxicity is of practical significance. The aim of this study was to examine the inhibitory effects of chitosan oligosaccharides (COS) on PM2.5-induced lung inflammation in rats. Forty SPF (specific pathogen-free) male Wistar rats weighing 200-220g were randomly divided into four groups: control group, COS group, PM2.5 group, and PM2.5+COS group. COS was pre-administered to rats by gavage at a single dose of 500mg/kg 2h before intratracheal instillation of PM2.5 at a single dose of 1.2mg/kg daily for 3 consecutive days. Normal saline (NS) was used as negative control. Twenty-four hours after the last instillation of PM2.5, rats were sacrificed and subjected to bronchoalveolar lavage (BAL). The BAL fluids (BALF) were collected for measurement of levels of total proteins, lactate dehydrogenase (LDH), interleukin-1 (IL-1β), IL-8, and tumor necrosis factor-ɑ (TNF-ɑ) using colorimetric or ELISA kits. Levels of total proteins, LDH activities, and pro-inflammatory mediators including IL-1β, IL-8, and TNF-ɑ in BALF of rats in PM2.5 group significantly increased in comparison with those of the control group. Pre-treatment of rats with COS markedly blocked PM2.5-induced increase in LDH, IL-8, and TNF-ɑ levels in BALF. In conclusion, PM2.5 exposure induces rat lung inflammation, which could be ameliorated by the pre-treatment of COS.

Role of the Rho/ROCK signaling pathway in the protective effects of fasudil against acute lung injury in septic rats.

Fasudil, which is primarily prescribed to treat cerebral vasospasm, may also inhibit systemic inflammation and prevent sepsis-induced acute lung injury (ALI) in rats, although the mechanisms remain elusive. The purpose of the present study was to investigate the role of the rhodopsin (Rho)/Rho-associated protein kinase (ROCK) signaling pathway in the protective effects of fasudil on ALI in septic rats. A total of 60 Wistar rats were pretreated with fasudil (30 mg/kg) through intraperitoneal injections 1 h prior to cecal ligation and puncture. Administration of fasudil led to reductions in polymorphonuclear neutrophil counts, and the protein concentrations of tumor necrosis factor-α, interleukin (IL)-1β and IL-6 in the bronchoalveolar lavage fluid of rats with sepsis-induced ALI. The results demonstrated that fasudil decreased sepsis-induced bacteremia. In addition, fasudil effectively reduced the Evans blue content, wet/dry lung weight ratio, lung injury score, and expression levels of malondialdehyde and myeloperoxidase. However, the superoxide dismutase activity in the lung tissue of the rats was increased. Activated caspase-3 activity in lung tissue was reduced to 29% by fasudil. Furthermore, the expression of Rho and ROCK1 was significantly downregulated, and the phosphorylation of myosin phosphatase-targeting subunit 1 in lung tissues was markedly decreased, whereas the protein expression levels of zonula occludens 1 were increased in fasudil-treated rats (P<0.05). In the in vitro experiments, vascular endothelial growth factor, intracellular adhesion molecule 1 and vascular cell adhesion molecule 1 secreted from human pulmonary microvascular endothelial cells treated with lipopolysaccharide (LPS) were attenuated by fasudil. Fasudil also reduced the fluorescence intensity of filamentous actin induced by LPS. Taken together, the results of the present study demonstrated that fasudil was able to improve endothelial permeability and inhibit inflammation, oxidative stress and cellular apoptosis in order to alleviate ALI in septic rats through inhibition of the Rho/ROCK signaling pathway.

Expression profile of microRNAs in bronchoalveolar lavage fluid of rats as predictors for smoke inhalation injury

Smoke inhalation injury (SII) is an independent risk factor for morbidity and mortality in patients with severe burns, however, the underlying mechanisms of SII are still not fully understood. In our study, we established an advanced rat model of SII based on the previous work, and explored the dynamic changes of pathophysiology and inflammatory factors during 28days post SII. We also measured the different expressions of miRNAs in bronchoalveolar lavage fluid (BALF) between SII and normal control rats by miRNA microarray. At 1day after smoke inhalation, the histopathological results exhibited inflammatory exudates in the lung tissue with significant edema. As time went on, the lung injuries gradually appeared at alveolar septum thickening and alveolar collapse, which suggested that it further induced damage to lung parenchyma by smoke inhalation. Particularly, the collagen deposition indicating pulmonary fibrosis happened at 28days post-injury. Plasma IL-6 and TNF-a were significantly increased after 1day of smoke inhalation. Plasma IL-10, BALF TNF-α and IL-10 were significantly increased after 2days of smoke inhalation. By extending the observation time, the levels of plasma IL-6, BALF TNF-a and IL-10 appeared a second peak again after 14days of injury. Compared with the normal control group, there were 23 upregulated miRNAs and 2 downregulated miRNAs in BALF of SII group at 1day post-injury. RT-qPCR validation assay confirmed that the changes of miR-34c-5p, miR-92b-3p, miR-205, miR-34b-3p, miR-92a-3p, let-7b-5p, let-7c-5p in BALF were consistent with the conclusion of the miRNA microarray. In summary, we showed the dynamic changes of pathologic changes and inflammatory factors in rats with SII, and a subset of seven miRNAs changed in BALF after SII which may be used for diagnosis and potential therapeutic targets.

NLRP3/ASC-mediated alveolar macrophage pyroptosis enhances HMGB1 secretion in acute lung injury induced by cardiopulmonary bypass

Our previous study showed that high levels of HMGB1 existed in rats following cardiopulmonary bypass (CPB)-induced acute lung injury (ALI) and neutralization of high-mobility group box 1(HMGB1) reduced CPB-induced ALI. However, the mechanism by which CPB increases HMGB1 secretion is unclear. Recent studies have shown that inflammasome-mediated cell pyroptosis promotes HMGB1 secretion. This study aimed to investigate the relationship between inflammasome-mediated pyroptosis and HMGB1 in CPB-related ALI. We employed oxygen-glucose deprivation (OGD)-induced alveolar macrophage (AM) NR8383 pyroptosis to measure HMGB1 secretion. We found that OGD significantly increased the levels of caspase-1 cleaved p10, IL-1β and ASC expression, caspase-1 activity and the frequency of pyroptotic AM, and promoted the cytoplasm transportation and secretion of HMGB1, which were significantly mitigated by ASC silencing or pre-treatment with glyburide (a Nlrp3 inhibitor) in AM. CPB also increased the expression levels of Nlrp3, ASC, caspase-1 P10, and IL-1β, and the percentages of AM pyroptosis in the lungs of experimental rats accompanied by increased levels of serum and bronchoalveolar lavage fluid (BALF) HMGB1. Treatment with glyburide significantly mitigated the CPB-increased ASC, caspase-1 p10 and IL-1β expression, and the percentages of AM pyroptosis in the lungs, as well as the levels of HMGB1 in serum and BALF in rats. Therefore, our data indicated that the Nlrp3/ASC-mediated AM pyroptosis increased HMGB1 secretion in ALI induced by CPB. These findings may provide a therapeutic strategy to reduce lung injury and inflammatory responses during CPB.Acute lung injury is a serious side effect of cardiopulmonary bypass. This study reveals that Nlrp3 inflammasome/ASC adaptor-mediated alveolar macrophage pyroptosis increases high-mobility group box 1 secretion in acute lung injury induced by cardiopulmonary bypass. These findings may provide a therapeutic strategy to reduce lung injury and inflammatory responses resulting from bypass surgery.

Alleviation of Acute Lung Injury in Rats with Sepsis by Resveratrol via the Phosphatidylinositol 3-Kinase/Nuclear Factor-Erythroid 2 Related Factor 2/Heme Oxygenase-1 (PI3K/Nrf2/HO-1) Pathway.

BackgroundResveratrol (Res) is a type of polyphenol found in many plants, which can protect important organs from the damage induced by sepsis. However, the exact mechanism of its protective effect has not been established. This study investigated the effect of Res on the PI3K/Nrf2/HO-1 signaling pathway in rats with sepsis-induced acute lung injury (ALI).Material/MethodsMale Wistar rats were treated with 30 mg/kg Res by intraperitoneal administration for 1 hour immediately after cecal ligation and puncture. Levels of MIP-2, IL-18, and IL-10 in bronchoalveolar lavage fluid (BALF) were determined. Lung tissues were collected to measure the wet-to-dry (W/D) ratios, oxidative stress index, and lung injury scores. Expression levels of Akt, p-Akt, HO-1, Nrf-2, and active caspase-3 proteins were determined by western blotting; expression of HO-1 mRNA was determined by RT-PCR.ResultsTreatment with Res significantly decreased the levels of MIP-2 and IL-18 and increased IL-10 in the BALF of rats with sepsis-induced ALI. In addition, Res also effectively reduced the W/D lung weight ratio, lung injury score, and the levels of MDA (malondialdehyde) and 8-OHdG. Conversely, Res increased SOD (superoxide dismutase) activity in the lung tissue. Moreover, Res significantly induced higher HO-1 mRNA expression, upregulated HO-1 and Nrf-2 protein expression, and the phosphorylation of Akt in the lung tissue. In contrast, the levels of activated caspase-3 protein were decreased in Res-treated rats (P<0.05).ConclusionsRes could inhibit inflammation, oxidative stress, and cell apoptosis to alleviate ALI in septic rats through the inhibition of the PI3K/Nrf2/HO-1 signaling pathway.

Endothelial Colony-forming Cells Attenuate Ventilator-induced Lung Injury in Rats with Acute Respiratory Distress Syndrome

Mechanical ventilation (MV) can cause ventilator-induced lung injury (VILI). This study investigated whether endothelial colony-forming cells (ECFC) could inhibit VILI in a rat model of acute respiratory distress syndrome (ARDS). Male Wistar rats received the femoral artery and venous cannulation (sham group) or were injected intravenously with 500μg/kg lipopolysaccharide to induce ARDS. The ARDS rats were subjected to MV. Immediately after the MV, the rats were randomized and injected intravenously with vehicle (ARDS group) or ECFC (ECFC group, n=8 per group). The oxygen index, lung wet-to-dry weight (W/D) ratios, cytokine protein levels in serum or bronchoalveolar lavage fluid (BALF), neutrophil counts, neutrophil elastase and total protein levels in BALF, histology and cell apoptosis in the lung were detected. The protein levels of endothelin-1, inducible nitric oxide synthase (iNOS), endothelial NOS, matrix metalloproteinase (MMP)-9, Bax, Bcl-2, gelsolin, cleaved caspase-3, phosphorylated NF-κBp65 and myosin light chain (MLC) in the lung were analyzed. Compared with the ARDS group, treatment with ECFC significantly increased the oxygen index, and decreased the lung W/D ratios and injury, and the numbers of apoptotic cells in the lungs, neutrophils counts, total protein and elastase concentrations in BALF of rats. ECFC treatment significantly minimized the protein levels of pro-inflammatory cytokines in BALF and serum, but increased interleukin 10 in rats. Furthermore, ECFC treatment significantly reduced the protein levels of endothelin-1, iNOS, Bax, Gelsolin, MMP-9, cleaved caspase-3, phosphorylated NF-κBp65 and MLC, but enhanced eNOS and Bcl-2 in the lungs of rats. Therefore, ECFC attenuated inflammation, cell apoptosis and VILI in ARDS rats.

Protective effects of tiotropium alone or combined with budesonide against cadmium inhalation induced acute neutrophilic pulmonary inflammation in rats.

As a potent bronchodilator, the anti-inflammatory effects of tiotropium and its interaction with budesonide against cadmium-induced acute pulmonary inflammation were investigated. Compared to values obtained in rats exposed to cadmium, cytological analysis indicated a significant decrease of total cell and neutrophil counts and protein concentration in bronchoalveolar lavage fluid (BALF) in rats pretreated with tiotropium (70μg/15ml or 350μg/15ml). Zymographic tests showed a decrease of MMP-2 activity in BALF in rats pretreated only with high concentration of tiotropium. Histological examination revealed a significant decrease of the severity and extent of inflammatory lung injuries in rats pretreated with both tested concentrations of tiotropium. Though tiotropium (70μg/15ml) or budesonide (250μg/15ml) could not reduce cadmium-induced bronchial hyper-responsiveness, their combination significantly decreased bronchial contractile response to methacholine. These two drugs separately decreased the neutrophil number and protein concentration in BALF but no significant interaction was observed when both drugs were combined. Although no inhibitory effects on MMP-2 and MMP-9 was observed in rats pretreated with budesonide alone, the combination with the ineffective dose of tiotropium induced a significant reduction on these parameters. The inhibitory effect of tiotropium on lung injuries was not influenced by budesonide which alone induced a limited action on the severity and extent of inflammatory sites. Our findings show that tiotropium exerts anti-inflammatory effects on cadmium-induced acute neutrophilic pulmonary inflammation. The combination of tiotropium with budesonide inhibits cadmium-induced inflammatory injuries with a synergistic interaction on MMP-2 and MMP-9 activity and airway hyper-responsiveness.

Interventional effects of BAY11-7082 on lung inflammatory response at the early stage and acute lung injury of rats with severe burns

Interventional effects of BAY11-7082 on lung inflammatory response at the early stage and acute lung injury of rats with severe burns

Effects of non-muscle myosin ⅡA silenced bone marrow mesenchymal stem cells on lung damage of rats at early stage of smoke inhalation injury

Objective: To investigate the effects of non-muscle myosin ⅡA (NMⅡA) silenced bone marrow mesenchymal stem cells (BMSCs) on the lung damage of rats at early stage of smoke inhalation injury. Methods: Forty Sprague-Dawley rats were divided into control, simple injury, NMⅡA-BMSCs, and BMSCs groups according to the completely random method, with 10 rats in each group. Rats in control group inhaled air normally, while rats in the latter 3 groups inhaled smoke to reproduce model of smoke inhalation injury. At 30 min post injury, rats in simple injury group were injected with 1 mL normal saline via caudal vein, and rats in group BMSCs were injected with 1 mL the fifth passage of BMSCs (1×10(7)/mL), and rats in group NMⅡA-BMSCs were injected with 1 mL NMⅡA silenced BMSCs (1×10(7)/mL). At post injury hour (PIH) 24, abdominal aorta blood and right lung of rats in each group were harvested, and then arterial partial pressure of oxygen (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)), and pH value were detected by blood gas analyzer. Ratio of wet to dry weight of lung was determined by dry-wet weight method. Pathological changes of lung were observed with HE staining. Bronchoalveolar lavage fluid (BALF) were collected, and then tumor necrotic factor-α (TNF-α) and interleukin-10 (IL-10) content of BALF was determined by enzyme-linked immunosorbent assay. Data were processed with one-way analysis of variance, Kruskal-Wallis H test, and least-significant difference test. Results: (1) At PIH 24, compared with those in control group, PaO(2) values of rats in simple injury, BMSCs, and NMⅡA-BMSCs groups were obviously decreased (with P values below 0.05), and PaCO(2) values were obviously increased (with P values below 0.05). Compared with those in simple injury group, PaO(2) values of rats in groups NMⅡA-BMSCs and BMSCs were obviously increased (with P values below 0.05), while PaCO(2) values were obviously decreased (with P values below 0.05). PaO(2) value of rats in group NMⅡA-BMSCs was obviously increased as compared with that in group BMSCs (P<0.05). The pH value of arterial blood of rats in simple injury group was obviously lower than that in control group (P<0.05). (2) At PIH 24, ratios of wet to dry weight of lung of rats in control, simple injury, BMSCs, and NMⅡA-BMSCs groups were 4.36±0.15, 7.79±0.42, 5.77±0.18, and 5.11±0.20, respectively. Compared with that in control group, ratio of wet to dry weight of lung of rats was obviously increased in the other 3 groups (with P values below 0.05). Compared with that in simple injury group, ratio of wet to dry weight of lung of rats was obviously decreased in groups BMSCs and NMⅡA-BMSCs (with P values below 0.05). Compared with that in group BMSCs, ratio of wet to dry weight of lung of rats in group NMⅡA-BMSCs was obviously decreased (P<0.05). (3) At PIH 24, alveolar structure of rats in control group was complete without abnormality. Compared with those in simple injury group, lung injury and infiltration of inflammatory cells of rats in groups BMSCs and NMⅡA-BMSCs were obviously alleviated, and alveolar structure was relatively complete with no thickening of alveolar wall. (4) At PIH 24, compared with that in control group, TNF-α content of BALF of rats in simple injury and BMSCs groups was obviously increased (with P values below 0.05). Compared with that in simple injury group, TNF-α content of BALF in groups BMSCs and NMⅡA-BMSCs was obviously decreased (with P values below 0.05). Compared with that in control group, IL-10 content of BALF in simple injury, NMⅡA-BMSCs and BMSCs groups were obviously increased (with P values below 0.05). Compared with that in simple injury group, IL-10 content of BALF in groups BMSCs and NMⅡA-BMSCs was obviously increased (with P values below 0.05). Compared with that in group BMSCs, IL-10 content of BALF in group NMⅡA-BMSCs was obviously increased (P<0.05). Conclusions: NMⅡA silenced BMSCs can alleviate lung damage of rats at early stage of smoke inhalation injury, showing better effectiveness than using BMSCs only.

The expression and activation of the AIM2 inflammasome correlates with inflammation and disease severity in patients with acute pancreatitis

This study explored the role and mechanism of salidroside in sepsis-associated acute lung injury (ALI).Sepsis-associated ALI in rats was induced by cecal ligation and perforation method, while lipopolysaccharide (LPS) was used to stimulate cell model. After the treatment of salidroside and dexamethasone (DEX), hematoxylin and eosin staining was applied to evaluate the rat lung injury. Next, the number of total cells and neutrophils in rat bronchoalveolar lavage fluid (BALF) was counted, and the lung wet/dry (W/D) ratio and water content were measured. The neutrophil elastase activity and inflammatory factor levels in BALF were detected by Elastase Assay kit and ELISA. The expressions of PPP1R15A and endoplasmic reticulum (ER) stress-related genes were detected by quantitative reverse transcription polymerase chain reaction and western blot. And the effects of silenced PPP1R15A and tauroursodeoxycholic acid (TUDCA) on cell viability, apoptosis and endoplasmic reticulum (ER) stress were determined by cell counting kit-8 assay, flow cytometry and western blot.The lung injury, inflammation and edema in sepsis model rats were alleviated by salidroside and DEX, meanwhile salidroside increased the viability and inhibited apoptosis in LPS-treated cells. The expression of PPP1R15A was decreased in sepsis models and increased by salidroside, and salidroside down-regulated the ER stress-related protein expressions in vitro and in vivo. Silenced PPP1R15A reversed the effect of salidroside on cell viability, apoptosis and ER stress, whereas TUDCA could counteract the above effect of silenced PPP1R15A.Salidroside targeted PPP1R15A to ameliorate lung injury in sepsis through inhibiting ER stress.

Effect of “three-acupoint and five-needling method” on the expression level of p-ERK 1/2 protein in lung tissue of rats with allergic bronchial asthma

ObjectiveTo explore the effect of “three-acupoint and five-needling method” on the expression level of p-ERK 1/2 protein in lung tissue of rats with allergic bronchial asthma. MethodsForty SPF-grade male SD rats were selected and randomly divided into four groups, including control group (group C), model group (group M), acupuncture group (group A) and blocker group (group PD), with 10 rats in each group. After corresponding intervention was given, cell counting was conducted, and the expression level of p-ERK 1/2 protein in the lung tissue of rats was detected via immunohistochemical method and Western blot method. ResultsThe number of leukocyte and eosinophil in the bronchoalveolar lavage fluid of rats in group A and group PD was smaller than that in group M, but larger than that in group C. It was shown based on the immunohistochemical test and Western blot test that the expression level of p-ERK1/2 protein in the lung tissue of rats in group M was higher than that in group C (P<0.05), while the expression level of p-ERK1/2 protein in group A and group PD was lower than that in group M (P<0.05). Conclusion“Three-acupoint and five-needling method” can effectively relieve the inflammatory response of rats with allergic bronchial asthma, and inhibit the expression level of p-ERK 1/2 protein in the lung tissue of rats with asthma. Acupuncture may alleviate the symptom of allergic bronchial asthma through inhibiting the expression level of p-ERK 1/2 protein.

Xuebijing injection attenuates pulmonary injury by reducing oxidative stress and proinflammatory damage in rats with heat stroke.

The present study aimed to investigate the protective effect of Xuebijing injection (XBJ) on lung injury in heat-stroke rats and the underlying mechanisms. In total, 54 rats were randomly assigned to non-thermal, saline vehicle and XBJ groups. The rectal temperature (Tc), mean arterial pressure (MAP) and respiratory rate (RR) of the rats were recorded. The time-point of heat stroke and the time of survival were assessed, and indicators of arterial blood gas were regularly measured from 0 to 60 min. The concentration of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-10 was also determined. At the end of the experiment, lung tissue was harvested for histopathological analysis. Inducible nitric oxide synthase (iNOS) and superoxide dismutase (SOD) expression was measured by immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to measure apoptosis. XBJ pretreatment prolonged the decline of clinical characteristics, as demonstrated by increases in Tc, MAP, RR and indicators in arterial blood gas in rats under heat stress. The time until heat stroke and the survival time in the Saline group were shorter than in rats treated with XBJ. The expression of iNOS in lung tissue and the concentration of TNF-α, IL-1β and IL-10 in the bronchoalveolar lavage fluid of rats treated with saline was higher than in rats with XBJ pre-treatment. Contrarily, SOD expression in rats treated with saline was decreased compared with that in rats treated with XBJ. Moreover, the apoptotic rate in the lung tissues of rats with saline treatment was higher than that in rats treated with XBJ. In conclusion, XBJ delayed the development of heat stroke and increased the survival time in rats under heat-stress by ameliorating pulmonary failure and acute lung injury. The underlying mechanisms of this effect may be the reduction of inflammatory cytokines as well as attenuation of oxidative stress and apoptosis by XBJ.

Analysis of Trace Elements in Rat Bronchoalveolar Lavage Fluid by Inductively Coupled Plasma Mass Spectrometry.

The main objective was to determine the elemental profile of the lung lining fluid of rats which are used as model animals in various experiments. Lung lining fluid elemental constitution obtained after bronchoalveolar lavage fluid (BALF) was analyzed by inductively coupled plasma mass spectrometry (ICP-MS) to determine the biological trace elements along with calcium and magnesium. BALF was collected from healthy rats using a tracheal cannula. However, cells in BALF were counted to monitor any underlying inflammatory lung condition. Cell free BALF samples were processed and analyzed for the elements including magnesium (Mg), calcium (Ca), chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), selenium (Se), bromine (Br), and iodine (I). In view of this, calcium concentration was the highest (6318.08±3094.3μg/L) and copper concentration was the lowest (0.89±0.21μg/L). The detected elements, from high to low concentration, include Ca>Mg>Fe>Br>I>Cr>Ni>Zn>Mn>Se>Cu. Pearson's correlation analysis revealed no significant correlation between cell count and concentration of any of the element detected in BALF. Correlation analysis also revealed significant positive correlation among Fe, I, Cr, Ni, and Mn. Ca was found to be correlated negatively with Cu and positively with Se. Br and Mg found to be positively correlated with each other. Zn remained the only element that was not found to be correlated with any of the elements in the rat BALF.

Comparative Proteomic Analysis of Rat Bronchoalveolar Lavage Fluid after Exposure to Zinc Oxide Nanoparticles.

Zinc oxide nanoparticles (ZnO NPs) are one of the most widely used nanomaterials in consumer products and industrial applications. As a result of all these uses, this has raised concerns regarding their potential toxicity. We previously found that candidate markers of idiopathic pulmonary fibrosis and lung cancer were significantly up-regulated in rat bronchoalveolar lavage fluid (BALF) following exposure to ZnO NPs by using a liquid chromatography (LC)-based proteomic approach. To achieve comprehensive protein identification analysis, we conducted the two-dimensional gel electrophosis (2-DE)-based proteomic workflow to analyze the differences in BALF proteins from rats that had been exposed to a high dose of 35 nm ZnO NPs. A total of 31 differentially expressed protein spots were excised from the gels and analyzed by nanoLC-tandem mass spectrometry (MS/MS). Gene ontology (GO) annotation of these proteins showed that most of the differentially expressed proteins were involved in response to stimulus and inflammatory response processes. Moreover, pulmonary surfactant-associated protein D and gelsolin, biomarkers of idiopathic pulmonary fibrosis, were significantly up-regulated in rat BALF after ZnO NPs exposure (2.42- and 2.84-fold, respectively). The results obtained from this present study could provide a complementary consequence with our previous study and contribute to a better understanding of the molecular mechanisms involved in ZnO NP-induced lung disorders.

Surfactant protein-A nanobody-conjugated liposomes loaded with methylprednisolone increase lung-targeting specificity and therapeutic effect for acute lung injury

The advent of nanomedicine requires novel delivery vehicles to actively target their site of action. Here, we demonstrate the development of lung-targeting drug-loaded liposomes and their efficacy, specificity and safety. Our study focuses on glucocorticoids methylprednisolone (MPS), a commonly used drug to treat lung injuries. The steroidal molecule was loaded into functionalized nano-sterically stabilized unilamellar liposomes (NSSLs). Targeting functionality was performed through conjugation of surfactant protein A (SPANb) nanobodies to form MPS–NSSLs–SPANb. MPS–NSSLs–SPANb exhibited good size distribution, morphology, and encapsulation efficiency. Animal experiments demonstrated the high specificity of MPS–NSSLs–SPANb to the lung. Treatment with MPS–NSSLs–SPANb reduced the levels of TNF-α, IL-8, and TGF-β1 in rat bronchoalveolar lavage fluid and the expression of NK-κB in the lung tissues, thereby alleviating lung injuries and increasing rat survival. The nanobody functionalized nanoparticles demonstrate superior performance to treat lung injury when compared to that of antibody functionalized systems.

Repair effect of Qinbai Qingfei concentrated pellets to AEC-Ⅱ of rats infected by Mycoplasmal pneumonia

To discuss the repair mechanism of Qinbai Qingfei concentrated pellets to AEC-Ⅱ of rats infected by mycoplasma through the observation of the changes and distribution of TGF-β and SP-A in lungs, totally 60 Wistar rats that weighing 80-100 g were collected, with male and female in half. The rats were divided into six groups randomly, with 10 rats each group, namely blank group, model group, positive group and Qinbai Qingfei concentrated pellets high, middle and low dose groups. Rats were infected through nasal intubation drip of MP. After 10 days of administration, serum and bronchoalveolar lavage fluid (BALF) were collected to detect the concentration of surface activity related protein A (SP-A) by ELISA, left pulmonary tissues of rats were collected to observe the expression and distribution of transforming growth factor-beta (TGF-β) and SP-A by immunohistochemistry, and right pulmonary tissues were taken to detect TGF-β and SP-A mRNA expression level by real-time quantitative PCR (RT-PCR). Qinbai Qingfei concentrated pellets can reduce the expression of TGF-β and increase the expression of SP-A in the lung tissues of rats infected by mycoplasma. Specifically, TGF-β was mainly distributed among the lung interstitium, while SPAs were mainly distributed in AEC-II and parts of alveolar macrophage. The level of SP-A was reduced in serum and increased in BALF in rats in Qinbai Qingfei concentrated pellets groups. It was proved that Qinbai Qingfei concentrated pellets can restore the normal morphology and function of the lung by reducing the content of TGF-β to inhibit epithelial-mesenchymal transition (EMT) of alveolar type II epithelial cells and increasing the expression of SP-A. Qinbai Qingfei concentrated pellets have the repairing ability to capillary vessel damage caused by MP in lung tissues of rats.