Adipose-derived mesenchymal stem cells (ADSCs) have demonstrated significant therapeutic effects on acute lung injury. Numerous studies have reported that preconditioning ADSCs can enhance their therapeutic efficacy. Currently, there is a lack of research on intermittent hypoxia preconditioning of ADSCs. In this study, we subjected ADSCs to intermittent hypoxic preconditioning, followed by the detection of their expressions of HIF-1α, HGF, VEGF, and IL-10. We also evaluated the therapeutic effects of ADSCs on oxidative stress and cell apoptosis in LPS-induced MLE-12 cells, and compared the outcomes resulting from different types of hypoxic preconditioning. Finally, we concluded that Intermittent hypoxia preconditioning leads to decrease in the secretion of VEGF by ADSCs compared to sustained hypoxia preconditioning; but their therapeutic effects in terms of anti-apoptosis and anti-oxidation are comparable.

Despite being a lifesaving intervention for many conditions, supplemental oxygen therapy carries the risk of severe hyperoxia-induced lung injury (HLI). The lack of effective countermeasures drives the urgent need for novel therapeutic strategies and mechanistic research to improve clinical outcomes. Eicosapentaenoic acid (EPA), a marine-derived polyunsaturated fatty acid with anti-inflammatory, antioxidant, and immunomodulatory properties, has exhibited lung-protective effects in experimental models of chronic obstructive pulmonary disease and bronchial asthma, suggesting its potential clinical value. However, its effects on HLI remain unexplored. This study investigated the therapeutic potential of EPA in alleviating HLI and elucidated its underlying mechanisms. We established both cellular and animal models of HLI; using these models, we integrated network pharmacology analysis to evaluate the efficacy of EPA and explore its potential mechanisms of action. Results demonstrated that EPA significantly ameliorated hyperoxia-induced oxidative stress, inflammatory responses, glycolytic reprogramming, and apoptosis in BEAS-2B and MLE-12 cells in vitro. Network pharmacology analysis revealed that EPA likely targets the MAPK/NF-κB signaling pathway. Western blotting and immunofluorescence confirmed that EPA attenuated hyperoxia-induced cellular damage by inhibiting p38 MAPK phosphorylation and reducing NF-κB nuclear translocation. In vivo, EPA suppressed the p38 MAPK/NF-κB signaling pathway, reduced inflammation and metabolic reprogramming in lung tissue, and thereby alleviated hyperoxia-induced pathological damage. This study substantiates that EPA ameliorates HLI by modulating glycolytic reprogramming via the p38 MAPK/NF-κB signaling pathway in vitro and in vivo, thereby providing a potential therapeutic strategy and laying the groundwork for its clinical application in HLI management.

BACKGROUND:Immunocompromise is common in the intensive care unit (ICU) and is strongly associated with adverse outcomes. However, robust quantitative tools for assessing the severity of immunocompromise are lacking. We aimed to develop and validate a machine learning-powered immunocompromise score and its risk stratification based on common immunocompromise conditions and biomarkers in the ICU. METHODS:A single-centre retrospective study was carried out in two ICUs of an academic tertiary care center in China. Adult patients who were admitted to the ICU for at least three days were enrolled. Feature selection was performed via the Boruta algorithm. The primary endpoint was 28-day all-cause mortality, whereas secondary endpoints included septic shock, the use of special antimicrobial agents, the peak levels of interleukin-6 (IL-6), etc. Seven machine learning models were developed via 10-fold cross-validation. The predicted probabilities from the optimal model were used to define the Immunocompromise and Severity (ICS) Score. To rapidly obtain patient immunocompromise information, a simplified ICS score (SICS score) was developed based on LASSO-selected features from day 1. Additionally, secondary endpoints were used to validate the rationale of ICS and SICS scores, and 216 patients were included for temporal validation. RESULTS:A total of 1863 patients were included for Algorithm derivation, with 679 deaths (36.9%). Among the seven machine learning models, the XGBoost model using data from the first 3 ICU days showed the highest performance, defined as ICS score (AUC 0.887; sensitivity 0.896; specificity 0.723; accuracy 0.787), and its performance was superior to that of the APACHE II and SOFA (P < 0.001). The LASSO algorithm selected 5 key variables (age, organ failure, immune impairing diseases and treatments, and IL-6 > 100 pg/mL with lymphopenia < 0.8 × 109/L) on ICU Day 1, for the SICS score (AUC 0.851; sensitivity 0.83; specificity 0.72; accuracy 0.765). The ICS and SICS scores effectively stratified patients into low, moderate, and high-risk groups with similar mortality rates (2.6% vs. 2.8%; 24.2% vs. 24.4%; 69.9% vs. 69.6%). Notably, the ICS score identified a greater proportion of patients in both the low (31.4% vs. 19.6%) and high risk (42.6% vs. 36.6%) groups. Secondary endpoints were significantly correlated with higher risk stratification, supporting the rationality of ICS and SICS scores. An additional 216 patients were used for temporal validation, yielding an AUC of 0.854 for the ICS score and 0.845 for the SICS score. CONCLUSIONS:Derived from common immunocompromising conditions and biomarkers, the ICS and SICS scores and their risk stratification system provide an accurate and timely solution for identifying and stratifying immunocompromise in critically ill patients. This framework may facilitate early clinical decision-making and resource allocation.

Background Phenotypic switching of vascular smooth muscle cells (VSMCs) serves as a critical pathological basis for various cardiovascular diseases. This phenotypic transformation enables cells to regain proliferative, secretory, and migratory capabilities, triggering pathological vascular remodeling and subsequent disease development, including pulmonary arterial hypertension (PAH). PAH represents a common complication of congenital heart disease (CHD). Beyond early surgical correction of anatomical defects, effective therapeutic options remain limited. Consequently, there is an urgent need to elucidate the mechanisms underlying PAH pathogenesis and identify novel therapeutic strategies. In CHD, pulmonary arteries are chronically exposed to a high-flow, hyperoxic environment. This study hypothesizes that chronic hyperoxia—a previously overlooked factor—may significantly contribute to phenotypic switching and functional alterations in pulmonary arterial smooth muscle cells (PASMCs). Methods PASMCs were isolated from Sprague-Dawley rats through enzymatic digestion and cultured under hyperoxic conditions to establish an in vitro hyperoxia model(95% O₂/5% CO₂). Cells were divided into two primary groups: Normoxia group (N-group) and Hyperoxia group (H-group). The effects of hyperoxia on phenotypic switching were assessed by examining the expression of contractile markers α-SMA and SM22α, and synthetic marker OPN using qRT-PCR and Western blot analysis. To evaluate changes in cellular secretory capacity, MMP-2 expression was analyzed at both mRNA and protein levels. Cell proliferation and migration capacities were evaluated using CCK-8 assays and scratch wound healing assays, respectively. To investigate the regulatory role of the PI3K/AKT pathway, cells were treated with the PI3K inhibitor LY294002 and divided into four experimental groups: Normoxia (N-group), Hyperoxia (H-group), Hyperoxia + LY294002 (HI-group), Normoxia + LY294002 (NI-group). The phenotypic and functional modifications of PASMCs were assessed using qRT-PCR, Western blot analysis, CCK-8 proliferation assays, and scratch wound healing migration assays to investigate the regulatory involvement of the PI3K/AKT signaling pathway. Results In PASMCs cultured under hyperoxic conditions, a phenotypic transition from contractile to synthetic state was observed. The expression of contractile proteins α-SMA and SM22α was downregulated, while the synthetic protein OPN was upregulated. Concurrently, hyperoxia induced elevated secretion of MMP-2, enhanced cellular viability as demonstrated by CCK-8 assay, and significantly augmented migratory capacity observed in wound healing assays. These alterations exhibited a time-dependent progression. Hyperoxia induced increased phosphorylation of PI3K and AKT, leading to activation of the PI3K/AKT signaling pathway. This activation was associated with enhanced cellular anti-apoptotic capacity and concomitant phenotypic and functional alterations Notably, treatment with the PI3K inhibitor LY294002 effectively abolished the hyperoxia-induced effects on both molecular profiles and cellular behavior. Conclusion The PI3K/AKT signaling pathway mediates hyperoxia-induced phenotypic transition in PASMCs, conferring enhanced proliferative, secretory, and migratory capacities. These findings establish a mechanistic link between chronic hyperoxia and pathological vascular remodeling, providing a novel pathological basis for CHD-associated PAH (CHD-PAH) development.

Background:Oxygen supplementation is essential for patients with a multitude of diseases but can cause severe hyperoxia-induced lung injury (HLI), necessitating the identification of therapeutic targets to improve clinical outcomes. Cuproptosis, a novel copper-dependent form of cell death characterized by proteotoxic stress resulting from lipoylated protein aggregation and loss of iron-sulfur cluster proteins, is distinct from other forms of cell death. However, the role of cuproptosis in HLI remains unclear. Methods:We established an HLI model in MLE-12 cells and C57BL/6 mice to investigate the involvement of cuproptosis in hyperoxia-induced toxicity. Results:We observed a time-dependent increase in the cuproptosis-related gene Fdx1 under hyperoxia. Moreover, hyperoxia activated the membrane-associated copper transporter SLC31A1 and significantly elevated copper levels in MLE-12 cells, as well as in the serum and lung tissue of C57BL/6 mice. Further analysis revealed that hyperoxia significantly altered the expression of cuproptosis-related genes without affecting DLAT levels, but significantly increased lipoylated-DLAT levels. ELISA, CCK-8 assays, HE staining, lung wet-to-dry weight ratio, and bronchoalveolar lavage fluid analysis demonstrated that treatment with the cuproptosis inhibitor TTM reduced pro-inflammatory cytokines (TNF-α and IL-1β) and alleviated hyperoxia-induced injury in both MLE-12 cells and C57BL/6 mice. Conclusion:Our study identifies the involvement of cuproptosis in HLI, providing new insights into the pathogenesis of hyperoxic lung injury and potential therapeutic strategies.

OBJECTIVES:To investigate the effect of overwork on myocardial energy metabolism in mice. METHODS:Thirty-two C57BL/6J mice were randomized equally into a control group and 3 overwork groups with overwork for 2, 4, and 6 weeks (W2, W4, and W6 groups, respectively). The mice in overwork groups were subjected to daily forced water standing and restraint. The changes in body weight and general condition of the mice were observed weekly. After successful modeling, the mice were examined for changes in echocardiography, blood glucose/lipid profiles, myocardial pathologies, myocardial TG and ATP levels, and expressions in CD36, GLUT1, CPT1B, PPARα, PFKM, and PKM2 using immunohistochemistry, RT-qPCR or Western blotting. RESULTS:The mice with prolonged overwork exhibited reduced activity with hair loss, dull fur, and slowed body weight gain without significant changes in cardiac index or function. Blood glucose levels increased significantly in W2 and W4 groups but decreased in W6 group. Serum TG level increased significantly while TC, HDL, and LDL decreased in W4 and W6 groups. HE staining revealed myocardial swelling, disorganization, and vacuolation in the mouse models. Myocardial TG was elevated in W4 and W6 groups and ATP level decreased in W6 group. The mRNA and protein expressions of CPT1B and PPARα were downregulated in W4 and W6 group, and CD36 expression increased significantly in W4 group. GLUT1 and PFKM/PKM2 expressions decreased obviously in W2 group but increased in W4 and W6 group compared with that in W2 group. CONCLUSIONS:Short-term overwork causes elevation of blood glucose and suppresses glycolysis in mice, while prolonged overwork reduces glucose, increases TG, impairs fatty acid oxidation, and limits glycolytic compensation to eventually result in myocardial damage, lipid accumulation, and ATP deficiency.

Oxygen therapy is widely used in pulmonary disease and critical care resuscitation. Unfortunately, prolonged exposure to high concentrations of oxygen may cause oxygen toxicity, leading to hyperoxia-induced acute lung injury (HALI). The injurious effects of lung following hyperoxia exposure are well established, and the effects include cell apoptosis and epithelial-to-mesenchymal transition (EMT). HALI responds to the inflammatory response induced by M1 macrophage polarization. Suppressing inflammation in macrophages protects against HALI. The PI3K/AKT pathway promotes cell survival in oxidative stress injury. Activation of Akt is a beneficial response protects against hyperoxic stress. FAM134B activates ER-phagy through binding to LC3B, ER fragments are degraded by ER-phagy, and ER homeostasis is maintained. We hypothesized that FAM134B may regulate macrophage polarization and inflammatory responses in hyperoxia-exposed rats via PI3K/AKT signaling pathway, thereby attenuating HALI. In this study, we clarified the role of FAM134B in lung tissues of hyperoxia-exposed rats and the effect of FAM134B on macrophage polarization. The expression of FAM134B and PI3K/AKT pathway were inhibited in rat lung tissues after hyperoxia exposure. We found that overexpression of FAM134B activated the PI3K/AKT pathway and reduced apoptosis and EMT in rat lung tissues after hyperoxia exposure. The PI3K/AKT pathway inhibitor, LY294002, reversed the protective effect of FAM134B in hyperoxia-exposed rats. Overexpression of FAM134B reduced the release of inflammatory factors (IL-1β, TNF-ɑ, IL-6) and polarization of M1 macrophages after hyperoxia exposure, and LY294002 reversed this effect. In conclusion, our study showed that FAM134B inhibits M1 macrophage polarization and inflammatory factor release via PI3K/AKT pathway and attenuates apoptosis and EMT in rat lung of hyperoxia-exposed FAM134B is a key target for HALI, which provides new ideas for the treatment of HALI.

Vital organ injury is one of the leading causes of global mortality and socio-economic burdens. Current treatments have limited efficacy, and new strategies are needed. Dexmedetomidine (DEX) is a highly selective α2-adrenergic receptor that protects multiple organs by reducing inflammation and preventing cell death. However, its exact mechanism is not yet fully understood. Understanding the underlying molecular mechanisms of its protective effects is crucial as it could provide a basis for designing highly targeted and more effective drugs. Ferroptosis is the primary mode of cell death during organ injury, and recent studies have shown that DEX can protect vital organs from this process. This review provides a detailed analysis of preclinical in vitro and in vivo studies and gains a better understanding of how DEX protects against vital organ injuries by inhibiting ferroptosis. Our findings suggest that DEX can potentially protect vital organs mainly by regulating iron metabolism and the antioxidant defense system. This is the first review that summarizes all evidence of ferroptosis's role in DEX's protective effects against vital organ injuries. Our work aims to provide new insights into organ therapy with DEX and accelerate its translation from the laboratory to clinical settings.

Objective To evaluate the effects of total intravenous anesthesia on the circadian rhythms in the patients undergoing cardiac transcatheter closure. Methods Thirty patients undergoing cardiac transcatheter closure under elective intravenous anesthesia were included in this study.Paired t-tests were performed to compare the mRNA levels of the genes encoding circadian locomotor output cycles kaput(CLOCK),brain and muscle ARNT-1 like protein-1(BMAL1),cryptochrome 1(CRY1),and period circadian clock 2(PER2),the Munich Chronotype Questionnaire(MCTQ)score,and the Pittsburgh Sleep Quality Index(PSQI)score before and after anesthesia.Multiple stepwise regression analysis was performed to screen the factors influencing sleep chronotype and PSQI total score one week after surgery. Results The postoperative mRNA level of CLOCK was higher [1.38±1.23 vs.1.90±1.47;MD(95%CI):0.52(0.20-0.84),t=3.327,P=0.002] and the postoperative mRNA levels of CRY1 [1.56±1.50 vs.1.13±0.98;MD(95%CI):-0.43(-0.81--0.05),t=-2.319,P=0.028] and PER2 [0.82±0.63 vs.0.50±0.31;MD(95%CI):-0.33(-0.53--0.12),t=-3.202,P=0.003] were lower than the preoperative levels.One week after surgery,the patients presented advanced sleep chronotype [3:03±0:59 vs.2:42±0:37;MD(95%CI):-21(-40--1),t=-2.172,P=0.038],shortened sleep latency [(67±64)min vs.(37±21)min;MD(95%CI):-30.33(-55.28--5.39),t=-2.487,P=0.019],lengthened sleep duration [(436±83)min vs.(499±83)min;MD(95%CI):62.80(26.93-98.67),t=3.581,P=0.001],increased sleep efficiency [(87.59±10.35)% vs.(92.98±4.27)%;MD(95%CI):5.39(1.21-9.58),t=2.636,P=0.013],decreased sleep quality score [1.13±0.78 vs.0.80±0.71;MD(95%CI):-0.33(-0.62--0.05),t=-2.408,P=0.023],and declined PSQI total score [6.60±3.17 vs.4.03±2.58;MD(95%CI):-2.57(-3.87--1.27),t=-4.039,P<0.001].Body mass index(BMI)(B=-227.460,SE=95.475,t=-2.382,P=0.025),anesthesia duration(B=-47.079,SE=18.506,t=-2.544,P=0.017),and mRNA level of PER2(B=2815.804,SE=1080.183,t=2.607,P=0.015)collectively influenced the sleep chronotype,and the amount of anesthesia medicine(B=0.067,SE=0.028,t=2.385,P=0.024)independently influenced the PSQI one week after surgery. Conclusion Total intravenous anesthesia can improve sleep habits by advancing sleep chronotype.BMI,anesthesia duration,and mRNA level of PER2 collectively influence sleep chronotype one week after surgery.The amount of anesthesia medicine independently influences the PSQI total score one week after surgery.

Objective To investigate the effect of overwork on the hemopoietic function of spleen in mice. Methods A total of 18 C57BL/6J mice were randomly divided into 3 groups, i.e., control group (C), 15-day-overworked group (W1) and 30-day-overworked group (W2). The mice from Groups W1 and W2 were forced to keep standing in water for 8 h and restrained for 3 h for consecutive 15 and 30 d, respectively, while those of Group C had no special treatment. The general condition, food intake and body weight of the mice were observed and measured. After modelling, the mice were euthanatized, and the spleen and heart were isolated to calculate spleen index and observe spleen pathological changes after HE staining, and then blood routine test was performed. The mRNA expression levels of CXCL12 and SCF in the spleen and that of connexin 45 (Cx45) in the heart were detected by RT-qPCR, and the protein levels of CXCL12 and SCF in spleen were detected by ELISA. The cardiac protein level of Cx45 was quantified with immunohistochemical assay. Results ① General condition showed that Groups W1 and W2 showed coarse hairs, increased at first and then decreased in activities, poor mental state, and larger food intake when compared with Group C. The body weight of Group C was increased with the feeding time, but that of Groups W1 and W2 had no such change. The spleen index was decreased significantly in Group W2 (P < 0.05). ② The results of blood routine test showed that WBC and RBC counts and HGB level were significantly increased in Group W2(P < 0.05), and PLT count in Groups W1 and W2 was significantly increased compared with Group C (P < 0.05). ③ Spleen HE staining displayed that the boundary between red pulp and white pulp was not clear, and the region of red pulp was enlarged in Group W2 when compared with Group C. ④ RT-qPCR showed that the mRNA level of Cx45 was significantly higher in heart tissue of Group W2 than that of Group C (P < 0.05), and that of SCF in spleen tissue of Group W2 was also higher than that in Groups C and W1 (P < 0.05). ⑤ Immunohistochemical assay indicated that the protein expression of Cx45 in myocardium of Groups W1 and W2 was significantly higher than that in Group C (P < 0.05). ⑥ ELISA revealed that the content of SCF in the spleen of Group W2 was notably higher than that of Groups C and W1 (P < 0.05), but there was no statistical difference between Groups W1 and C. Conclusion Overwork destroys the tight connection between cardiac myocytes, enhances the hemopoietic function of spleen, increases WBC count, and then forms a hypercoagulable state, which may be one of the precipitating factors for cardiovascular diseases related to overwork.

Oxygen therapy is widely used in clinical practice; however, prolonged hyperoxia exposure may result in hyperoxic acute lung injury (HALI). In this study, we investigated the role of FAM134B in hyperoxia-induced apoptosis, cell proliferation, and epithelial-to-mesenchymal transition (EMT) using RLE-6TN cells and rat lungs. We also studied the effect of CeO2-NPs on RLE-6TN cells and lungs following hyperoxia exposure. FAM134B was inhibited in RLE-6TN cells and rat lungs following hyperoxia exposure. Overexpressing FAM134B promoted cell proliferation, and reduced EMT and apoptosis following hyperoxia exposure. FAM134B activation increased ER-phagy, decreased apoptosis, improved lung structure damage, and decreased collagen fiber deposition to limit lung injury. These effects could be reversed by PI3K/AKT pathway inhibitor LY294002. Additionally, CeO2-NPs protected RLE-6TN cells and lung damage following hyperoxia exposure by ameliorating impaired ER-phagy. Therefore, FAM134B restoration is a potential therapeutic target for the HALI. Moreover, CeO2-NPs can be used for the treatment of HALI.

Interstitial lung disease and pulmonary sarcoidosis remain serious medical problems worldwide. This study aims to assess the global burden and health inequalities of interstitial lung disease and pulmonary sarcoidosis between 1990 and 2021. Data on disability-adjusted life years (DALYs) due to interstitial lung disease and pulmonary sarcoidosis were obtained from the Global Burden of Diseases, Injuries and Risk Factors Study 2021. The slope index of inequality (SII) and concentration index were used to assess cross-national health inequality. There were 2,237,269 (95

Objective To observe the effect of excess oxygen supply for different time periods on the mitochondrial energy metabolism in alveolar epithelial type Ⅱ cells. Methods Rat RLE-6TN cells were assigned into a control group (21% O2 for 4 h) and excess oxygen supply groups (95% O2 for 1,2,3,and 4 h,res-pectively).The content of adenosine triphosphate (ATP),the activity of mitochondrial respiratory chain complex V,and the mitochondrial membrane potential were determined by luciferase assay,micro-assay,and fluorescent probe JC-1,respectively.Real-time fluorescence quantitative PCR was employed to determine the mRNA levels of NADH dehydrogenase subunit 1 (ND1),cytochrome b (Cytb),cytochrome C oxidase subunit I (COXI),and adenosine triphosphatase 6 (ATPase6) in the core subunits of mitochondrial respiratory chain complexes Ⅰ,Ⅲ,Ⅳ,and Ⅴ,respectively. Results Compared with the control group,excess oxygen supply for 1,2,3,and 4 h down-regulated the mRNA levels of ND1 (q=24.800,P<0.001;q=13.650,P<0.001;q=9.869,P<0.001;q=20.700,P<0.001),COXI (q=16.750,P<0.001;q=10.120,P<0.001;q=8.476,P<0.001;q=14.060,P<0.001),and ATPase6 (q=22.770,P<0.001;q=15.540,P<0.001;q=12.870,P<0.001;q=18.160,P<0.001).Moreover,excess oxygen supply for 1 h and 4 h decreased the ATPase activity (q=9.435,P<0.001;q=11.230,P<0.001) and ATP content (q=5.615,P=0.007;q=5.029,P=0.005).The excess oxygen supply for 2 h and 3 h did not cause significant changes in ATPase activity (q=0.156,P=0.914;q=3.197,P=0.116) and ATP content (q=0.859,P=0.557;q=1.273,P=0.652).There was no significant difference in mitochondrial membrane potential among the groups (F=0.303,P=0.869). Conclusion Short-term excess oxygen supply down-regulates the expression of the core subunits of mitochondrial respiratory chain complexes and reduces the activity of ATPase,leading to the energy metabolism disorder of alveolar epithelial type Ⅱ cells.

沉默信息调节因子1(SIRT1)作为一种去乙酰化酶,可以脱去多种靶蛋白赖氨酸残基的乙酰基.近年来,越来越多的研究证实SIRT1在肠道疾病的发病机制中占有重要的地位.因此,SIRT1可能成为治疗肠道疾病的潜在靶点.本文综述了SIRT1对氧化应激,炎症与细胞凋亡等生物效应的调控作用;归纳总结了SIRT1在脓毒症肠损伤,肠缺血再灌注损伤,放射性肠损伤,肠道炎症性疾病等疾病中的变化和作用,以及靶向SIRT1治疗肠道疾病的药物研究进展,旨在探讨肠道疾病新的防治策略.

Objective To explore the effect of overwork (OW) on extracellular matrix of arterial vessel wall in rats. Methods Random number grouping method was employed to assign 18 Sprague-Dawley rats into three groups(n=6):the control group(no special treatment),group OW(forced swimming twice a day for 15 days),and sleep deficiency(SD)+OW group(in addition to forced swimming twice a day,the rats were put on the platforms in water to limit sleep for 15 days).On the 16th day,the abdominal aorta and common carotid artery were collected after blood sampling from heart under deep anesthesia.A part of the abdominal aorta sample was taken for Masson staining of collagen fiber,and Verhoeff-Van Gieson staining was carried out for the elastic fiber of common carotid artery.Image J was employed for the quantitative analysis of collagen fiber and elastic fiber content.The expression of collagen 1(Col-1) protein was quantified by immunohistochemistry and the ultrastructure of vascular matrix was examined by transmission electron microscopy.The other part of the abdominal aorta sample was used to determine the mRNA levels of matrix metalloproteinase(MMP)-1,MMP-2,MMP-9,tissue inhibitor of metalloproteinases-1(TIMP-1),and Col-1 by quantitative real-time polymerase chain reaction. Results Compared with that in control group,the content of collagen fiber in groups OW and SD+OW had no significant change(all P>0.05);the content of elastic fiber in groups OW and SD+OW decreased(all P<0.001) and had no significant difference between each other(P>0.05).The vascular vessel wall of group OW showed slight fiber breakage,while that of group SD+OW presented wormhole-like or spongy fiber fragmentation.The mRNA levels of MMP-1 and MMP-2 in groups OW and SD+OW had no significant difference between each other(P>0.05) but were higher than that in control group(all P<0.001).The mRNA levels of MMP-9 and TIMP-1 had no significant difference among the three groups(all P>0.05).Groups OW and SD+OW had lower mRNA level(all P<0.001) and protein level(all P<0.001) of Col-1 than control group,while the mRNA and protein levels of Col-1 had no significant difference between groups OW and SD+OW(P>0.05). Conclusion OW can reduce the content of Col-1 and elastic fibers in the extracellular matrix of arterial vessels,destroy the elastic lamina of vascular wall,up-regulate the expression of MMP-1 and MMP-2,thereby injuring arterial vessels.

目的 探讨富氢水(hydrogen-rich water)对高氧环境下小鼠肠道屏障和菌群的影响.方法 选取SPF级C57BL/6雄性小鼠24只,随机分为4组(n=6):对照组(C组)、对照+富氢水组(CH组)、高氧组(H组)和高氧+富氢水组(HH组).C组、CH组小鼠饲养于常氧环境,H组、HH组小鼠饲养于85％高氧环境.CH组和HH组小鼠每天给予富氢水0.1mL/10g灌胃2次,持续7d;C组和H组小鼠给予等体积生理盐水.第7天采集小鼠粪便,心脏采血,处死动物并切取末端回肠组织5 cm.HE染色观察肠组织病理学变化,行肠黏膜损伤评分;透射电镜观察肠上皮细胞超微结构;测定肠组织丙二醛(MDA)水平,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力,肠分泌型免疫球蛋白A(sIgA)水平;测定血清二胺氧化酶(DAO)水平;提取粪便DNA行肠道菌群检测和分类.结果 与C组、CH组小鼠相比,H组肠黏膜损伤评分升高,肠道SOD、CAT活力下降,sIgA、MDA水平升高,血清DAO水平升高(均P＜0.05);与H组小鼠相比,HH组肠黏膜损伤评分下降,MDA水平下降,肠道SOD、CAT活力升高,血清DAO水平下降(均P＜0.05).基因测序分析发现4组小鼠间肠道菌群多样性差异无统计学意义(P＞0.05),但菌群的组成有明显差异:H组小鼠肠道菌群F16科、梭状芽胞杆菌属、葡萄球菌属和理研菌属富集;HH组小鼠肠道菌群蓝细菌门、双歧杆菌属和阿克曼菌属富集.结论 高氧7d后小鼠肠屏障功能受到氧化应激损伤,并影响了肠道菌群组成.富氢水可减轻高氧肠损伤,其机制与上调抗氧化酶SOD、CAT活力,降低氧化应激产物MDA水平,保持肠道机械屏障完整和肠道有益菌富集有关.

Objective To determine the effect of overworking on vascular smooth muscle cells (VSMC) in rats. Methods Clean SD rats were randomly divided into control group (CON), overworking group (OW) and overworking and sleep deficiency group (OW+SD). The rat model of overworking was established by exhaustive swimming in the latter 2 groups, and the rats of the last group was subjected to sleep deprivation with an underwater platform. In 15 d later, the abdominal aorta of rats was collected for pathological observation by HE staining, and the thickness of media was measured with aid of Image J software. The intracellular ultrastructure was observed by transmission electron microscopy. The apoptotic rate of VSMC was detected by TUNEL, the mRNA expression of Caspsae-3, Bax, Bcl-2 and Ki67 in the VSMC were detected by RT-qPCR, and the content of apoptotic protein Caspase-3 was measured by immunohistochemical assay. Results ① The results of HE staining showed that VSMC were in disordered arrangment and formed vacuole-like cells in the OW and OW+SD groups, and the vascular intima-media thickness was significantly thicker in the OW and OW+SD groups than the CON group (P < 0.05). ② Electron microscopy displayed the apoptotic bodies in the VSMC of the OW and OW+SD groups. ③TUNEL results showed that the apoptotic rate of VSMC was significantly higher in the OW and OW+SD groups than the CON group (P < 0.05). ④RT-qPCR showed that the expression levels of Bax and Ki67 were significantly increased (P < 0.05), while those of Caspase-3 and Bcl-2 were not obviously changed in OW and OW+SD groups (P>0.05) when compared with the CON group. ⑤ Immunohistochemical results showed that the protein content of Caspase-3 in vascular middle membrane of the OW and OW+SD groups was increased significantly than that in the CON group (P < 0.05). Conclusion Overworking aggravates the apoptosis of VSMC and induces the proliferation and repair of VSMC, which form the pathological basis of early arterial lesions. During the experimental time, sleep could not alleviate the injury of VSMC caused by overworking.

目的 研究富氢水(HRW)通过调节沉默信息调节因子1/核因子E2相关因子2/血红素加氧酶-1(SIRT1/Nrf2/HO-1)通路对小鼠高氧肠损伤发挥保护作用的机制.方法 C57BL/6小鼠24只,随机分为常氧组(N组)、高氧组(O组)和富氢水组(H组)和富氢水联合SIRT1抑制剂组(HE组),每组6只.N组小鼠置于室内环境(FiO2=21％)中,O组、H组和HE组小鼠置于高氧环境(FiO2=85％)中,持续7 d.H组给予0.1 mL/10 g富氢水灌胃,2次/d,连续7 d;HE组在H组基础上每天腹腔注射SIRT1抑制剂EX52710 mg/kg 1次,连续7 d;N组和O组给予等体积生理盐水.每天称质量并记录小鼠体质量,7 d后安乐死小鼠并留取标本.取回肠组织制备切片并行Chiu病理评分,透射电镜观察回肠组织超微结构,检测回肠组织超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,Western blot检测回肠组织SIRT1、Nrf2、HO-1表达水平.结果 与N组比较,O组、H组、HE组实验后小鼠体质量减轻,Chiu病理评分、MDA含量升高,SOD活性降低;与O组比较,H组实验后小鼠体质量增加,H组、HE组Chiu病理评分、MDA含量降低SOD活性升高;与H组比较,HE组实验后小鼠体质量减轻,Chiu病理评分、MDA含量升高,SOD活性降低[体质量(g):N组23.17±1.70,O组17.60±1.58,H组20.87±1.24,HE组17.20±1.27;Chiu病理评分(分):N组0.33±0.27,O组3.44±0.37,H组2.11±0.25,HE组3.06±0.23;MDA含量(nmol/mg):N组59.15±3.77,O组86.18±3.88,H组68.12±2.27,HE组76.94±5.26;SOD活性(U/mg):N组105.51±7.67,O组64.58±4.62,H组86.45±5.51,HE组72.43±4.27;均P<0.05].与N组比较,O组SIRT1表达下调,Nrf2、HO-1表达上调,H组、HE组SIRT1、Nrf2、HO-1表达上调;与O组比较,H组SIRT1、Nrf2、HO-1表达上调,HE组SIRT1、Nrf2表达上调;与H组比较,HE组SIRT1、Nrf2、HO-1表达下调(SIRT1:N组0.17±0.01,O组0.13±0.01,H组0.66±0.04,HE组0.36±0.01;Nrf2:N组0.07±0.01,O组0.23±0.02,H组0.54±0.02,HE组0.38±0.02;HO-1:N组0.37±0.01,O组0.42±0.02,H组0.56±0.02,HE组0.44±0.02;均P<0.05).结论 富氢水预处理可通过激活SIRT1/Nrf2/HO-1信息通路,对小鼠高氧肠损伤发挥保护作用.

Background Long-term mechanical ventilation with hyperoxia can induce lung injury. General anesthesia is associated with a very high incidence of hyperoxaemia, despite it usually lasts for a relatively short period of time. It remains unclear whether short-term mechanical ventilation with hyperoxia has an adverse impact on or cause injury to the lungs. The present study aimed to assess whether short-term mechanical ventilation with hyperoxia may cause lung injury in rats and whether deferoxamine (DFO), a ferrous ion chelator, could mitigate such injury to the lungs and explore the possible mechanism. Methods Twenty-four SD rats were randomly divided into 3 groups ( n = 8/group): mechanical ventilated with normoxia group (MV group, FiO 2 = 21%), with hyperoxia group (HMV group, FiO 2 = 90%), or with hyperoxia + DFO group (HMV + DFO group, FiO 2 = 90%). Mechanical ventilation under different oxygen concentrations was given for 4 h, and ECG was monitored. The HMV + DFO group received continuous intravenous infusion of DFO at 50 mg•kg − 1 •h − 1 , while the MV and HMV groups received an equal volume of normal saline. Carotid artery cannulation was carried out to monitor the blood gas parameters under mechanical ventilation for 2 and 4 h, respectively, and the PaO 2 /FiO 2 ratio was calculated. After 4 h ventilation, the right anterior lobe of the lung and bronchoalveolar lavage fluid from the right lung was sampled for pathological and biochemical assays. Results PaO 2 in the HMV and HMV + DFO groups were significantly higher, but the PaO 2 /FiO 2 ratio were significantly lower than those of the MV group (all p < 0.01), while PaO 2 and PaO 2 /FiO 2 ratio between HMV + DFO and HMV groups did not differ significantly. The lung pathological scores and the wet-to-dry weight ratio (W/D) in the HMV and HMV + DFO groups were significantly higher than those of the MV group, but the lung pathological score and the W/D ratio were reduced by DFO ( p < 0.05, HMV + DFO vs. HMV). Biochemically, HMV resulted in significant reductions in Surfactant protein C (SP-C), Surfactant protein D (SP-D), and Glutathion reductase (GR) levels and elevation of xanthine oxidase (XOD) in both the Bronchoalveolar lavage fluid and the lung tissue homogenate, and all these changes were prevented or significantly reverted by DFO. Conclusions Mechanical ventilation with hyperoxia for 4 h induced oxidative injury of the lungs, accompanied by a dramatic reduction in the concentrations of SP-C and SP-D. DFO could mitigate such injury by lowering XOD activity and elevating GR activity.