Upregulation of PGC-1α expression by pioglitazone mediates prevention of sepsis-induced acute lung injury

Abstract The imbalance between pro-inflammatory M1 and anti-inflammatory M2 macrophages plays a critical role in the pathogenesis of sepsis-induced acute lung injury (ALI). Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) may modulate macrophage polarization toward the M2 phenotype by altering mitochondrial activity. This study aimed to investigate the role of the PGC-1α agonist pioglitazone (PGZ) in modulating sepsis-induced ALI. A mouse model of sepsis-induced ALI was established using cecal ligation and puncture (CLP). An in vitro model was created by stimulating MH-S cells with lipopolysaccharide (LPS). qRT-PCR was used to measure mRNA levels of M1 markers iNOS and MHC-II and M2 markers Arg1 and CD206 to evaluate macrophage polarization. Western blotting detected expression of peroxisome proliferator-activated receptor gamma (PPARγ) PGC-1α, and mitochondrial biogenesis proteins NRF1, NRF2, and mtTFA. To assess mitochondrial content and function, reactive oxygen species levels were detected by dihydroethidium staining, and mitochondrial DNA copy number was measured by qRT-PCR. In the CLP-induced ALI mouse model, lung tissues exhibited reduced PGC-1α expression. PGZ treatment rescued PGC-1α expression and alleviated lung injury, as evidenced by decreased lung wet-to-dry weight ratio, pro-inflammatory cytokine secretion (tumor necrosis factor-α, interleukin-1β, interleukin-6), and enhanced M2 macrophage polarization. Mechanistic investigations revealed that PGZ activated the PPARγ/PGC-1α/mitochondrial protection pathway to prevent sepsis-induced ALI by inhibiting M1 macrophage polarization. These results may provide new insights and evidence for developing PGZ as a potential ALI therapy.

Edema pulmonar por reexpansión. Reporte de Caso / Pulmonary edema due to re-expansion. Case Report

Introducción. El edema pulmonar por reexpansión es una complicación poco frecuente, secundaria a una rápida reexpansión pulmonar posterior al drenaje por toracentesis o toracostomía cerrada. Al día de hoy, se ha descrito una incidencia menor al 1 % tras toracostomía cerrada, con mayor prevalencia en la segunda y tercera década de la vida. Su mecanismo fisiopatológico exacto es desconocido; se ha planteado un proceso multifactorial de daño intersticial pulmonar asociado con un desequilibrio de las fuerzas hidrostáticas. Caso clínico. Presentamos el caso de un paciente que desarrolló edema pulmonar por reexpansión posterior a toracostomía cerrada. Se hizo una revisión de la literatura sobre esta complicación. Resultados. Aunque la clínica sugiere el diagnóstico, la secuencia de imágenes desempeña un papel fundamental. En la mayoría de los casos suele ser autolimitado, por lo que su manejo es principalmente de soporte; sin embargo, se han reportado tasas de mortalidad que alcanzan hasta el 20 %, por tanto, es importante conocer los factores de riesgo y las medidas preventivas. Conclusión. El edema pulmonar de reexpansión posterior a toracostomía es una complicación rara en los casos con neumotórax, aunque es una complicación que se puede presentar en la práctica diaria, por lo cual debe tenerse en mente para poder hacer el diagnóstico y un manejo adecuado.

Introduction. Re-expansion pulmonary edema is a rare complication secondary to rapid pulmonary re-expansion after drainage by thoracentesis and/or closed thoracostomy. As of today, an incidence of less than 1% has been described after closed thoracostomy, with a higher prevalence in the second and third decades of life. Its exact pathophysiological mechanism is unknown; a multifactorial process of lung interstitial damage associated with an imbalance of hydrostatic forces has been proposed. Clinical case. We present the case of a patient who developed pulmonary edema due to re-expansion after closed thoracostomy, conducting a review of the literature on this complication. Results. Although the clinic suggests the diagnosis, the sequence of images plays a fundamental role. In most cases, it tends to be a self-limited disease, so its management is mainly supportive. However, mortality rates of up to 20% have been recorded. Therefore, it is important to identify patients with major risk factors and initiate preventive measures in these patients. Conclusions. Re-expansion pulmonary edema after thoracostomy is a rare complication in cases with pneumothorax; however, it is a complication that can occur in daily practice. Therefore, it must be kept in mind to be able to make the diagnosis and an adequate management.

Bibliometric and visual analysis of SCI indexed papers on e-cigarette or vaping product use associated lung injury from 2013 to 2022 / 环境与职业医学

Background In recent years, the rising number of e-cigarette users among adolescents and the surging cases of lung injury related to e-cigarette use have attracted the attention of researchers in various fields. Objective To identify the research hotspots and trends of e-cigarette or vaping product use associated lung injury (EVALI) worldwide from 2013 to 2022 by bibliometric and visual analysis. Methods Web of Science Core Collection was selected to obtain literature related to EVALI from 2013 to 2022 across the world, statistics were calculated by country/region, institution, author, journal, cited literature, keyword, etc. CiteSpace 6.2.R1 was used for visual analysis to draw diagrams of publication trend, author cooperation network, co-citation clustering time distribution, and keyword cluster. Results A total of 888 EVALI-related papers published between 2013 and 2022 were retrieved. The number of publications was gradually increased, with a significant increase in 2020 and a decrease from 2021, but the number of citations was increased year by year. The most active country was the United States (631 articles). European and American countries cooperated closely and the centrality was prominent. Among the publishing institutions, the University of California system topped the list with 103 articles. Rahman I (27 articles) published the most articles and had a high degree of centrality; Goniewicz M L was the most cited author; and the network analysis diagram showed relatively weak collaboration between authors. The American Journal of Respiratory and Critical Care Medicine was the journal with the highest number of publications (94 articles). The top 5 cited articles were all cited more than 300 times. The leading high-frequency keywords of EVALI-related studies were nicotine (149 times), exposure (118 times), and oxidative stress (80 times). The cluster of key nodes in the co-citation network and the clustering time distribution diagram indicated youth e-cigarette addiction received widespread attention from society. From the top 25 keywords with the strongest bursts, the focus of research on the pathogenesis of EVALI gradually shifted from the oxidative stress damage associated with e-cigarette vapor to the oxidative effect of flavoring chemicals in the process of lung injury. The current research interests in this field were mainly the mechanisms of various chemicals in e-cigarettes and the heating elements that led to damage to the lungs. Conclusion EVALI is receiving continuous attention from researchers in government, medical institutions, and other organizations. A variety of e-cigarette ingredients such as flavoring chemicals may lead to varying degrees of cytotoxicity, inflammation, and lung damage. However, the pathophysiology of EVALI remains unclear. In the future, more Chinese scholars should be encouraged to participate in this field.

Effect and mechanism of cucurbitacin B preventing sepsis-induced acute lung injury in mice / 中国药房

OBJECTIVE To investigate the preventive effect of cucurbitacin B （CB） on sepsis-induced acute lung injury （ALI） and its mechanism. METHODS The mice were divided into control group， model group， dexamethasone group （positive control， 2 mg/kg）， CB low-dose and high-dose groups （25， 50 mg/kg）. Each group was given relevant medicine intraperitoneally， once a day， for 3 consecutive days. Twenty-four hours after the last administration， those groups were given lipopolysaccharide （10 mg/kg） intraperitoneally to establish sepsis-induced ALI model （finally， 8 mice per group were included in the experiment）， except for control group. Twenty-four hours after medication， blood routine indicators （total white blood cell count， neutrophils count， lymphocytes count）， lung function indicators （total lung resistance， pulmonary outflow resistance， lung dynamic compliance， peak expiratory flow rate， and maximum ventilation volume）， dry wet ratio of lung tissue were measured in each group. The lung tissue level of myeloperoxidase （MPO）， and the serum levels of tumor necrosis factor-α （TNF-α）， interleukin 1β （IL-1β）， IL-6， superoxide dismutase （SOD） and malondialdehyde （MDA） were all detected. The pathological changes of lung tissue were observed； immunohistochemistry was used to detect the positive expression of phosphorylation signal transducer and activator of transcription 3 （p-STAT3） in the lung tissue. Western blot assay was used to detect the expressions of proteins related to IL-6/JAK2/ STAT3 signaling pathway in the lung tissue. RESULTS Compared with control group， total pulmonary resistance， pulmonary flow resistance， dry wet ratio of lung tissue， the total white blood cell count， neutrophils count， lymphocytes count of whole blood， the lung tissue level of MPO and serum levels of MDA， IL-6， IL-1β and TNF-α， the p-STAT3 protein optical density value， the protein expressions of IL-6 and IL-6 receptor， and the phosphorylation levels of JAK2 and STAT3 protein were increased significantly in the model group （P＜0.01）， while lung dynamic compliance， peak expiratory flow rate， maximum ventilation volume and serum level of SOD were decreased significantly （P＜0.05 or P＜0.01）. Pulmonary tissue showed alveolar collapse and infiltration of inflammatory cells. Compared with the model group， the above indexes of mice were reversed significantly in dexamethasone group and CB groups （P＜0.05 or P＜0.01）， the pathological damage of lung tissue was reduced. CONCLUSIONS CB can prevent sepsis-induced ALI by inhibiting the activity of Δ 基金项目遵义市科技计划项目（No.202252） IL-6/JAK2/STAT3 signaling pathway and relieving ＊第一作者主治医师。研究方向：重症医学。E-mail：fjuanxui@ 163.com inflammatory reactions. # 通信作者 主任医师。研究方向：儿童呼吸系统疾病诊断与治

HMGB1 gene knockout alleviates acute lung injury of sepsis mice via inhibiting TLR4/NF-κB pathway / 安徽医科大学学报

Objective To study the effect of high mobility group box B1(HMGB1)gene knockout on alleviating a-cute lung injury and inhibiting toll-like receptor 4(TLR4)/nuclear factor-KB(NF-κB)pathway of sepsis mice.Methods Wild-type(WT)mice were divided into WT-Sham group and WT-model group,and HMGB1 knockout(KO)mice were divided into KO-sham group and KO-model group.Sepsis ALI model was established by cecal ligation and perforation in WT-model group and KO-model group.Sham operation was performed in WT-Sham group and KO-Sham group.24 h after modeling,the partial pressure of arterial oxygen(PaO2)was detected,oxy-genation index(OI)was calculated,pathological changes of lung tissue were detected and lung injury score was calculated,the concentrations of tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1 β),interleukin-6(IL-6),reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD),in serum and lung tissues and the expression of HMGB1,TLR4 and nuclear NF-κB in lung tissues were detected.Results The PaO2,OI and the concentration of SOD in serum and lung tissue of WT-model group were lower than those of WT-Sham group,the lung injury scores,the concentrations of TNF-α,IL-1 β,IL-6,ROS and MDA in serum and lung tissue,and the expression levels of HMGB1,TLR4 and nuclear NF-κB in lung tissue were higher than those in WT-Sham group(P＜0.05).HMGB1 was not expressed in lung tissue of KO-model group,and the concentrations of PaO2,OI and the concentration of SOD in serum and lung tissue of KO-model group were higher than those of WT-model group,the lung injury scores,the concentrations of TNF-α,IL-1β,IL-6,ROS and MDA in serum and lung tissue,and the expression levels of TLR4 and nuclear NF-κB in lung tissue were lower than those of the WT-model group(P＜0.05).Conclusion HMGB1 gene knockout alleviates acute lung injury of sepsis mice,the re-lated molecular mechanism may be the inhibition of TLR4/NF-κB pathway mediated inflammation and oxidative stress.

Effects and mechanism of methionine restriction on macrophage for lipopolysaccharide-induced acute lung injury in mice / 陆军军医大学学报

Objective To investigate the effects of methionine restriction(MR)on macrophages in lipopolysaccharide(LPS)-induced acute lung injury(ALI)and to explore the underlying mechanism.Methods According to the random number table method,36 male C57BL/6J mice(6～8 weeks old,23±2 g)were divided into 3 groups with 12 mice in each group:the sham group,the LPS group and the LPS+MR group.HE staining and pathological scoring of lung injury were performed in lung tissues.The expression of LPS-binding protein(LBP)and Toll-like receptor-4(TLR4)was detected by RT-qPCR and Western blotting.Macrophage-colony stimulating factor(M-CSF),granulocyte-macrophage-colony stimulating factor(GM-CSF)and chemokine C-C motif ligand 3(CCL3)which are all macrophage-associated chemokines were analyzed by immunohistochemistry.Results Compared with the sham group,the pathological score of lung injury in the LPS group was significantly increased(P＜0.01);The mRNA and protein expression levels of LBP and TLR4 were significantly increased;The number of positive cells of CD11b,F4/80,M-CSF,GM-CSF and CCL3 were significantly increased(P＜0.01).MR significantly improved LPS-induced ALI,and decreased the pathological score of lung injury(P＜0.01);The mRNA and protein expression levels of LBP and TLR4 were decreased;Compared with the LPS group,the number of positive cells of CD11 b,F4/80,M-CSF,GM-CSF and CCL3 were reduced in the LPS+MR group(P＜0.01).Conclusion MR could attenuate LPS-induced ALI by inhibiting the expression of macrophage chemokines and preventing infiltration and activation of macrophage to lungs.

Evaluation value of serum AGEs level combined with LIS score on prognosis of elderly patients with sepsis-related ALI/ARDS / 国际检验医学杂志

Objective To study the evaluation value of lung injury score(LIS)and advanced glycation end products(AGEs)expression levels on the prognosis of elderly patients with sepsis-related acute lung injury/acute respiratory distress syndrome(ALI/ARDS).Methods A total of 98 elderly patients with sepsis-related ALI/ARDS admitted to First Branch of the First Affiliated Hospital of Chongqing Medical University from March 2019 to April 2021 were selected as the research group,and the patients were divided into two sub-groups according to their survival within 30 d after admission:the survival group(55 cases)and the death group(43 cases).Another 51 elderly patients with non-ALI/ARDS sepsis admitted to First Branch of the First Affiliated Hospital of Chongqing Medical University in the same period were selected as the control group.After admission,the clinical data of patients were recorded,and the levels of serum creatinine,troponin I,B-type brain natriuretic peptide(BNP),serum C-reactive protein(CRP)and procalcitonin(PCT)were de-tected.Enzyme-linked immunosorbent assay was used to determine the levels of AGEs in patients'serum.The LIS score was evaluated by LIS scale.With clinical factors as independent variables and prognosis as dependent variables,Logistic regression curve was used to analyze the death factors of elderly sepsis-related ALI/ARDS patients.Results AGEs levels,LIS scores,acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)scores decreased sequentially in the death group,survival group,and control group(all P<0.05).The levels of lactic acid,blood glucose,troponin I,PCT,BNP and CRP in arterial blood of patients in the death group were significantly higher than those in the survival group and the control group(P<0.05).The results showed that arterial lactate,blood glucose,troponin I,PCT,BNP,CRP,AGEs,APACHE Ⅱ score,and LIS score were all independent risk factors for mortality in elderly sepsis-related ALI/ARDS patients(P<0.05).The area under the curve(AUC)of LIS score predicting prognosis in elderly sepsis-related ALI/ARDS pa-tients was 0.857(95%CI:0.821-0.911),and AUC of serum AGEs was 0.861(95%CI:0.809-0.908).LIS score and AGEs level had certain predictive value for the prognosis of elderly sepsis-related ALI/ARDS pa-tients.Conclusion The LIS score and AGEs level of the elderly patients with sepsis-related ALI/ARDS are independent risk factors of death,which have important predictive value for prognosis.

Improvement of sepsis-related acute lung injury through Naringin by regulating TGF-β1/Smad2 signaling pathway / 国际中医中药杂志

Objective:To investigate the protective effect of naringenin on acute lung injury related with sepsis; To discuss its possible mechanism.Methods:Totally 30 male SD rats were randomly divided into sham-operation group, model group, naringin low-, medium- and high-dosage groups, with 6 rats in each group. The sepsis-related acute lung injury model was established by cecal ligation and puncture in all groups except the sham-operation group. After modeling, naringin low-, medium- and high-dosage groups were given naringin 20 mg/kg, 40 mg/kg and 80 mg/kg, respectively for gavage, while the sham-operation group and the model group were given the same volume of distilled water by gavage, once a day, for 2 days. Pathological changes in lung tissue were observed using HE staining. The levels of 1L-1, IL-6 and IL-18 in bronchoalveolar lavage fluid (BALF) were measured by ELISA; the expression of TNF-α in lung tissue was detected by immunofluorescence histopathology; the expressions of TGF-β1, TGF-βR1 and Smad2 were detected by Western Blot. An agonist group and a naringin plus agonist group were set up, with 6 mice in each group, and the expressions of TGF-β1 and Smad2 protein in the lung tissue of each group were detected by immunohistochemical staining to verify the effect of naringin on the expressions of TGF-β1 and Smad2 protein.Results:Compared with the model group, the pathological injury of lung tissue in naringin groups were obviously alleviated, and the levels of IL-1β, IL-6 and IL-18 in BALF decreased ( P<0.01), the protein expressions of TNF-α, TGF-β1, TGF-βR1 and Smad2 in lung tissue decreased ( P<0.01 or P<0.05). Further verification found that the expressions of TGF-β1 and Smad2 in the agonist group increased ( P<0.01), while the expressions of TGF-β1 and Smad2 in the naringin agonist group decreased ( P<0.01). Conclusion:Naringin can reduce the inflammatory response in the lung of the rats to protect against sepsis-related acute lung injury, and its protective effect could be related to the inhibition of the TGF-β1/Smad2 signaling pathway.

Macrophage-to-myofibroblast transition promotes pulmonary fibrosis occurred in LPS-induced acute lung injury of mouse models / 基础医学与临床

Objective To explore the impact of macrophage-to-myofibroblast transition(MMT)on pulmonary fibro-sis induced by acute lung injury by LPS.Methods Totally 21 male mice were randomly classified into 7 groups:control group,model group(LPS-PF)at different time points and intervention group of clodronate-liposomes(CL-LIP)treatement at different time points(n=3).Pulmonary fibrosis was identified by HE and Masson staining microscopy.The immuno-fluorescence technology was used for the evaluation of numbers of macrophage-to-myofi-broblast transition cells(MMT cell which co-expressed CD68 and α-SMA).Bone marrow-derived macrophages(BMDMs)were randomly classified into two group:control(Ctrl)group and TGF-β1-treated group induced by transforming growthfactor-β1.α-SMA,FN and Col1 were detected by RT-qPCR.The expression of α-SMA,Smad3 and p-Smad3 protein was evaluated by Western blot.Results At day 7,the Ashcroft score of lung tissue in LPS-PF mouse model was significantly increased when compared with the Ctrl group(P＜0.01);While the score signifi-cantly declined when the model was pretreated with CL-LIP(P＜0.05).As detected by immuno-fluorescence stai-ning,in CL-LIP group the number of CD68-positive cells co-labeled with α-SMA was obviously less then that of LPS-PF group of the corresponding time point(P＜0.01).When the BMDMs were stimulated by TGF-β1 at 24 h,48 h and 96 h respectively,a higher expression of α-SMA,FN,Col1,were found in TGF-β1-treated group than that in Ctrl group at the corresponding time point(P＜0.01).The expression of Smad3,p-Smad3 significantly higher in LPS-PF group(at both day 7 and day 10)and TGF-β1-treated group(at both 48 h and 96 h)as compared to cor-responding control group(P＜0.01).Conclusions MMT promotes pulmonary fibrosis induced by ALI via LPS.Smad3 is proved to be involved in the MMT process.

Dexmedetomidine alleviates lung tissue injury of rat models with ventilator-associated lung injury / 基础医学与临床

Objective To investigate the effect of dexmedetom idine(DEX)on lung tissue and Ras homolog gene family member A(RhoA)/Rho kinase 1(ROCK1)signaling pathway in lung tissue of rats with ventilator-induced lung injury(VILI).Methods A VILI rat model was established and separated into control group,model group(VILI group),dexmedetomidine low and high dose groups(DEX-L,DEX-H group),and high dose dexmedetomi-dine+lysophosphatidic acid(LPA)group(DEX-H+LPA group).Determination of wet/dry mass ratio of rat lung tissue(W/D);HE staining microscopy was applied to observe morphology of lung tissue;ELISA kit was applied to detect the level of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in bronchoalveolar lavage fluid(BALF);TUNEL staining method was applied to detect lung epithelial cell death;Immunoblotting was applied to detect the expression levels of apoptosis-related proteins,and RhoA,ROCK1 pro-teins.Results DEX could reduce lung injury,lung injury score,W/D,apoptosis rate,levels of TNF-α,IL-1β,IL-6,and expression of Bax,cleaved caspase-3,RhoA,ROCK,α-SMA in VILI rats(P＜0.05),while increased the expression of Bcl-2(P＜0.05);LPA could aggravate lung injury and increase lung injury score,W/D,apopto-sis rate,level of TNF-α,IL-1β,IL-6 and expressions of Bax,cleaved caspase-3,RhoA,ROCK and α-SMA(P＜0.05);Bcl-2 expression level was decreased(P＜0.05).Conclusions Dexmedetomidine may protect rats with ventilator-induced lung injury by the inhibition of RhoA/ROCK1 signaling pathway.

Glabridine regulates NETs to inhibit pyrodeath and alleviate lung injury in sepsis / 中华急诊医学杂志

Objective:To investigate the effect of glabridin on neutrophil extracellular traps (NETs) formation and pyroptosis in rats with sepsis-induced lung injury.Methods:Twenty-four male Wistar rats were divided into three groups according to the random number table method. The sepsis group was established by cecal ligation and puncture (CLP). The Glabridin group underwent CLP and glabridin gavage (30 mg/kg)(CLP+GLA). The sham operation group underwent cecal exploration, and only the abdomen was closed after cecal turning(Sham). After 12 hours, plasma、alveolar lavage fluid and lung tissue samples were taken for detection . Then, protein content of the alveolar lavage fluid was determined; The wet/dry weight(W/D) ratio of the lung tissue was determined; The pathological changes in lung tissue were observed after hematoxylin-eosin (HE) staining. The levels of NETs marker MPO-DNA complex and related inflammatory factors IL-18 and IL-1β in plasma were detected by enzyme-linked immunosorbent assay. The changes of Caspase-1and Cleaved-caspase-1 protein in lung tissue were detected by Western blot.Results:The total protein concentration of alveolar lavage fluid was significantly higher in the sepsis group compared with the Sham group ( P<0.01), and it decreased in the glabridin group compared with the sepsis group ( P<0. 05). Significant aggravation of pulmonary edema in the sepsis group, and the glabridin group reduced pulmonary edema compared with the sepsis group.The pathological results of lung tissue under the light microscope showed: The structure of lung tissue in the Sham group was normal, and the alveoli were clear; In the sepsis group, the alveolar wall was thickened widely and inflammatory cells infiltrated obviously; Compared with the sepsis group, the lung tissue injury was significantly reduced in the light licorice group. The enzyme-linked immunosorbent assay results showed that the levels of NETs marker MPO-DNA complex and inflammatory factors IL-18 and IL-1β in the plasma of the sepsis group were significantly higher than those in the Sham group ( P<0.001). The levels of NETs marker MPO-DNA complex and inflammatory factors IL-18 and IL-1β in the glabridin group were significantly lower than those in the sepsis group (MPO-DNA: P<0. 01; IL-18、IL-1β: P<0.05) . Western blot Technical results showed that the expression of Caspase-1 and Cleaved-caspase-1 protein positive signal was significantly enhanced in the lung tissue of the rats in the sepsis group compared with the Sham group; the distribution of Caspase-1 positive cells in the lung tissue of the sepsis + glabridin group was similar to that of the Sham group, and the expression of Cleaved-caspase-1 positive signal was higher than that of the Sham group. Conclusions:Glabridin can effectively reduce lung inflammation and play a protective role in lung injury in septic rats by inhibiting NETs production and pyroptosis.

Effect of Shenfu injection on acute lung injury in hemorrhagic shock rats / 临床麻醉学杂志

Objective To observe the effect of Shenfu injection on lung injury caused by hemor-rhagic shock(HS)in rats and explore the related potential mechanism.Methods Thirty-six SPF healthy male SD rats,aged 16-17 weeks,weighing 400-600 g,were randomly divided into three groups:sham op-eration group(group SH),HS group(group HS),and Shenfu injection group(group SF),12 rats in each group.In group SH,only the right femoral vein and femoral artery were separated after anesthesia,and ve-nous catheterization was not performed.HS model was established in groups SF and HS.In group HS,liquid resuscitation was performed through an intravenous catheter,and the resuscitation fluid consisted of the auto-blood lost and the compound sodium chloride injection equivalent to 1.5 times the blood loss and 10 ml/kg normal saline.In group SF,the resuscitation fluid consisted of the lost autoblood and the compound sodium chloride injection equivalent to 1.5 times the blood loss and Shenfu injection 10 ml/kg.The whole perfusion time was about 60 minutes.Six rats in the three groups were randomly anesthetized 24 and 48 hours after op-eration.The wet/dry weight ratio(W/D)of lung tissues was detected.The concentrations of interleukin-6(IL-6),IL-17,IL-10,and transforming growth factor-β(TGF-β)were detected by ELISA,the mRNA ex-pression of retinoic acid-related orphan nuclear receptor γt(RORγt),transcription factor forkhead box pro-tein 3(Foxp3),and hypoxia-inducible factor-1α(HIF-1α)in lung tissues were detected by PCR.The pro-tein contents of RORγt,Foxp3,HIF-1α,aquaporin 1(AQP1),and AQP5 in lung tissue were detected by Western blot.Pathological changesunder HE staining light microscope and lung injury scores were observed.Results Compared with 24 hours after operation,W/D,the concentrations of IL-6 and IL-17,mRNA ex-pression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concentrations of IL-10,and TGF-β,Foxp3 mRNA expression and protein content,and AQP1 protein content were significantly increased in group SF 48 hours after operation(P＜0.05).Compared with group SH,W/D,the concentrations of IL-6,IL-17,IL-10,and TGF-β,mRNA expression and protein content of RORγt,Foxp 3,and HIF-1α,and lung injury score were significantly increased(P＜0.05),AQP1 and AQP5 protein contents were significantly decreased in groups HS and SF 24 and 48 hours after operation(P＜0.05),and alveolar structure was damaged under light microscope and alveolar interstitium was filled with a large amount of edematous fluid,during which a large number of inflammatory cells infiltra-ted.Compared with group HS,W/D,the concentrations of IL-6 and IL-17,mRNA expression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concen-trations of IL-10 and TGF-β,Foxp3 mRNA expression and protein content,AQP1 and AQP5 protein con-tents were significantly increased in group SF 24 and 48 hours after surgery(P＜0.05),and the alveolar structure was improved under light microscope,and edema was reduced,and the number of inflammatory cells was reduced.Conclusion Shenfu injection can regulate the balance between pro-inflammatory factors IL-6 and IL-17,and anti-inflammatory factors IL-10 and TGF-β,increase the protein content of AQP1 and AQP5 in lung tissue,and decrease the W/D and injury score in lung tissue,thus alleviating lung injury in HS rats.The mechanism may be related to the regulation of HIF-1α-RORγt/Foxp3 balance.

Study on the Expression Levels and Clinical Value of Serum CCL25 and PARK7 in Patients with Acute Lung Injury Secondary to Sepsis / 现代检验医学杂志

Objective To investigate the serum levels of C-C motif chemokine ligand 25(CCL25)and human Parkinson's disease protein 7(PARK7)in patients with sepsis and their relationship with acute lung injury(ALI).Methods 138 sepsis patients diagnosed and treated in Hefei Jingdongfang Hospital from February 2019 to February 2023 were selected as sepsis group.They were divided into ALI group(n=40)and non-ALI group(n=98)based on whether ALI occurred.70 healthy individuals who underwent physical examinations at the same time were taken as a control group.Enzyme linked immunosorbent assay was used to detect serum levels of CCL25 and PARK7.The correlation between serum CCL25,PARK7 and clinical indicators were analyzed by Pearson correlation analysis.Risk factors for secondary ALI in sepsis were conducted by multivariate logistic regression analysis.The value of serum CCL25 and PARK7 levels in predicting secondary ALI in sepsis were conducted by the receiver operating characteristic curve.Results Serum CCL25(367.52±46.87ng/L)and PARK7(54.26±17.45μg/L)in patients with sepsis was higher than that of the control group(48.17±5.26ng/L,12.31±4.12 μg/L),and the differences were statistically significant(t=46.825,19.813,all P＜0.05).ALI group patients CCL25(434.65±52.87ng/L vs 340.12±42.64ng/L),PARK7(103.47±22.51μg/L vs 34.18±7.46 μg/L),respiratory index(1.58±0.48 vs 0.88±0.07),PaCO2(50.11±6.27mmHg vs 40.42±5.20mmHg),APACHE Ⅱ score(23.37±3.82 point vs 17.15±3.41 point)and SOFA score(13.56±2.93 point vs 10.18±2.81 point)were all higher in the non-ALI group,while oxygenation index(237.14±23.56 point vs 341.14±21.37 point)and PaO2(55.87±8.03mmHg vs 63.11±7.14mmHg)were lower in the non-ALI group,and the differences were statistically significant(t=10.998,27.151,14.145,9.342,9.385,6.332,25.172,5.210,all P＜0.05).The serum levels of CCL25 and PARK7 in ALI patients were positively correlated with APACHE II score,SOFA score,respiratory index and PaCO2(r=0.579～0.801,all P＜0.05),while negatively correlated with oxygenation index and PaO2(r=-0.687,-0.643;-0.654,-0.712,all P＜0.05).Serum CCL25(OR=1.309,95%CI:1.040～1.646),PARK7(OR=1.288,95%CI:1.016～1.633),APACHE II score(OR=1.188,95%CI:1.019～1.384)and SOFA score(OR=1.197,95%CI:1.006～1.425)were independent risk factors for secondary ALI in sepsis patients.The area under the curve(95%CI)of the combination of serum CCL25 and PARK7 for predicting secondary ALI in sepsis was 0.833(0.784～0.872),which was greater than the individual indicators 0.770(0.725～0.835)and 0.741(0.691～0.790),and the differences were statistically significant(Z=4.602,4.318,P＜0.05).Conclusion Elevated serum levels of CCL25 and PARK7 in patients with sepsis are independent risk factors affecting the occurrence of secondary ALI in sepsis.The combination of the two has high predictive value for secondary ALI in sepsis.

Hydroxynonenal alleviates neonatal sepsis-induced acute lung injury by inhibiting endothelial cell pyrosis / 实用医学杂志

Objective To explore the role of 4-hydroxynonenal(HNE)in alleviating acute lung injury(ALI)induced by neonatal sepsis by inhibiting the focal death of endothelial cells(ECs).Methods Newborn mice were randomly divided into five groups:(1)Sham operation group(Sham group),(2)sham operation mice receiving HNE treatment group(Sham + HNE group),(3)cecal serosity(CS group),and(4)CS-treated GS-DMD-/-mice group(CS + GSDMD-/-group).The degree of lung injury was evaluated by lung histopathology and lung wet/dry weight ratio.The ECs of mice were isolated and divided into the Ctrl group,LPS + ATP group,LPS + ATP + HNE-L group and LPS + ATP + HNE-H group.Western blot was used to evaluate the expression of HNE and caspase-1 pathway.Results Compared with CS group,the lung tissue scores of CS + HNE group and CS + GSDMD-/-group were significantly decreased(P<0.05),and the ratio of wet to dry weight of lung tissues was significantly decreased(P<0.05).Compared with the CS group,the 72-hour survival rates of mice in the CS + HNE group and CS + GSDMD-/-group were significantly improved(P<0.05).The expressions of GSDMD-N,C-caspase-1,NLRP3,IL-18 and IL-1β in lung ECs of the CS + HNE group and CS + GSDMD-/-group were signifi-cantly lower than those of the CS group(P<005).Compared with the Ctrl cells,LPS + ATP significantly decreased the cell viability(P<0.05)and increased the protein expressions of GSDMD,C-caspase-1,NLRP3,IL-18 and IL-1β(P<0.05),and these effects were also inhibited by HNE.Conclusion HNE can inhibit the focal death of lung ECs cells by inhibiting NLRP3/caspase-1 signal transduction,and improve ALI in septic mice.

Codonopsis polysaccharide protected LPS-induced acute lung injury by inhibiting MAPK/NF-κB signaling pathway in mice / 实用医学杂志

Objective To observe the protective effects of codonopsis pilosulae polysaccharide on lung tissues in mice with acute lung injury(ALI)induced by lipopolysaccharide(LPS)and its mechanism.Methods Fifty male Kunming mice were randomly divided into control group,model group,dexamethasone group,codonopsis polysaccharide high-dose group(300 mg/kg)and codonopsis polysaccharide low-dose group(150 mg/kg).The ALI model was established by intraperitoneal injection of LPS.All mice were given gavage administration according to the grouping except for the control group.0.3 s force expiratory volume(FEV 0.3)and force spirometry(FVC)and their ratios were measured using multiparametric lung function test system.The histopathology change of mouse lung was detected by hematoxylin-eosin(HE)staining,and the classification and count of inflammatory cells in alveolar lavage fluid(BALF)was detected by Richter-Giemsa staining.Levels of IL-1β,IL-6,MPO and TNF-α were measured by ELISA in BALF,and Western blot was used to detect the protein expression level of p-p38,p-IκB-α and p-p65.Results Compared with those in the control group,lung histopathological damage was more pronounced in the model mice,with significantly lower FEV 0.3,FVC,FEV0.3/FVC assay value,but signifi-cantly higher lung tissue wet mass/dry mass(W/D),neutrophils,lymphocytes,IL-1β,IL-6,MPO,TNF-α,p-p38 MAPK,p-IκB-α,and p-p65(P<0.05);while codonopsis pilosulae polysaccharide could significantly reverse these effects.Conclusion Codonopsis pilosulae polysaccharide plays a protective role against LPS-induced ALI via inhibiting MAPK/NF-κB pathway to reduce the pathological damage of lung tissue,and the level of inflammatory factors,thus to improve lung function in ALI model mice.

Effect of phillyrin regulating NLRP3 inflammatory pathway on exudates and lung injury in rats with acute pleurisy / 天津医药

Objective To investigate the impacts of phillyrin on exudates and lung injury in rats with acute pleurisy by regulating the NLRP3 inflammatory pathway.Methods Ninety rats were randomly divided into the control group,the model group,the low-dose phillyrin(PH-L,5 mg/kg)group,the medium-dose phillyrin(PH-M,10 mg/kg)group,the high-dose phillyrin(PH-H,20 mg/kg)group and the NLRP3 pathway inhibitor(PJ34,10 mg/kg)group.FVC,FEV 0.1 and FEV 0.3 were detected by lung function analyzer.Electronic balance was used to weigh the mass of chest exudate.The number of white blood cells in exudate was detected by Wright staining.Contents of prostaglandin E2(PGE2),monocyte chemoattractant protein-1(MCP-1),interleukin(IL)-6 and tumor necrosis factor-α(TNF-α)in exudate were detected by ELISA.Automatic blood gas analyzer was used to detect p(CO2)and p(O2)of rats.HE staining was used to observe pathological changes of lung tissue.The expression levels of NLRP3 and Caspase-1 protein were detected by immunohistochemistry.Western blot assay was used to detect the expression of NLRP3 pathway protein.Results Compared with the control group,the quality of pleural exudate and the number of white blood cells,the contents of PGE2,MCP-1,IL-6,TNF-α,the expression of p(CO2)and NLRP3 pathway proteins in exudate of the model group increased obviously,FVC,FEV 0.1,FEV 0.3 and p(O2)decreased obviously,and the lung tissue showed obvious pathological damage(P＜0.05).Compared with the model group,the quality of pleural exudate and the number of white blood cells,the contents of PGE2,MCP-1,IL-6,TNF-α,the expression of p(CO2),NLRP3 pathway proteins in the exudate of rats decreased obviously in the PH group and the PJ34 group,FVC,FEV 0.1,FEV 0.3 and p(O2)increased obviously,the pathological injury of lung tissue was obviously improved(P＜0.05).Compared with the PH-H group,there were no significant differences in the above indexes in the PJ34 group(P＞0.05).Conclusion PH can improve lung injury induced by acute pleurisy in rats by inhibiting the activation of NLRP3 pathway and inhibiting inflammatory reaction.

Jinyinqingre Oral Liquid Alleviates Acute Lung Injury in Mice through NF-κB/NLRP3 Signaling Pathway / 医药导报

Objective To explore the protective effect and mechanism of Jinyinqingre oral liquid on acute lung injury in-duced by lipopolysaccharide(LPS)in mice.Methods C57BL/6J mice were randomly divided into six groups according to the random number table method:blank control group,model control group,Jinyinqingre oral liquid low-dose group,Jinyinqingre oral liquid medium-dose group,Jinyinqingre oral liquid high-dose group,and dexamethasone group.Except for the blank control group,the other groups were injected with lipopolysac charide(LPS)(5 mg·kg-1)into the trachea to establish the acute lung injury model of mice,and the Jinyinqingre oral liquid low,medium,and high groups were continuously administered the drug by gavage for three days.After 24 h,lung tissue and bronchoalveolar lavage fluid(BALF)were collected from the six groups for follow-up detection.The pathological injury of lung tissue in each group was observed by HE staining.The total number of cells in BALF was detected.The to-tal protein content of BALF was detected by the BCA method.The contents of inflammatory cytokines TNF-α,IL-1β,IL-6,and IgM in BALF were detected by ELISA.The expression of NF-κB and NLRP3 proteins in lung tissues was detected by immunohistochemistry and Western blotting.Results Compared with model control group,Jinyinqingre oral liquid alleviated the pathological injury of lung tissue(P＜0.05),decreased the total cell count,total protein content and IgM expression in BALF(P＜0.01),and the expres-sion of inflammatory cytokines TNF-α,IL-6 and IL-1β in BALF was dreased(P＜0.05),the protein expressions of NF-κB and NL-RP3 in lung tissues was dreased(P＜0.05).Conclusion Jinyinqingre oral liquid attenuated the pathological injury,inflammatory exudation,and expression of inflammatory cytokines in LPS-induced lung injury in mice,and its mechanism may be through the reg-ulation of NF-κB/NLRP3 signaling pathway,providing a theoretical basis for its clinical application.

Characterization and correlation analysis of lung flora in rats with silica-induced acute lung injury / 中国病理生理杂志

AIM:To elucidate the possible biological mechanism of silica-induced acute lung injury in rats.METHODS:Sixteen Male Sprague-Dawley rats were divided into control and acute silicosis model groups,and instilled intratracheally with 1 mL of normal saline and 50 g/L silica suspension,respectively.After 7 d,the rats were sacrificed for collection of lung tissue and serum.The serum levels of interleukin-1β(IL-1β),IL-18 and tumor necrosis factor-α(TNF-α)were measured by using ELISA.The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and gasdermin D(GSDMD)were measured by immunohistochemistry.Bacterial DNA was ex-tracted from the lung tissue for 16S ribosomal RNA gene sequencing to characterize changes in the composition of lung flo-ra.The differences in the structure of bacterial flora between control and model groups were analyzed by bioinformatic analy-ses.RESULTS:Immunohistochemical analysis showed that the protein expression levels of NLRP3 and GSDMD were higher in the lungs of the rats in model group.In addition,serum cytokine profiling showed that IL-1β,IL-18 and TNF-α levels were significantly higher in model group.The most abundant bacterial genera in the lung flora of the rats in model group were Bifidobacterium,Clostridium sensu stricto 1,and Parasutterella.The NLRP3 and GSDMD levels in the lung tissue and IL-1β and TNF-α levels in serum were positively correlated with the abundance of Parasutterella.CONCLU-SION:The alterations in lung flora structure and increased inflammation levels may be the actual biological mechanisms underlying silica-induced acute lung injury.The modulation of lung flora may provide a basis for the prevention and treat-ment of silica-induced acute lung injury.

Saponin from Panax japonicus IVa alleviates lipopolysaccharide-induced acute lung injury in rats via its anti-inflammatory and antioxidative ef-fects / 中国病理生理杂志

AIM:To investigate the effects of saponin from Panax japonicus IVa(SPJ IVa)on acute lung inju-ry in rats and to explore its possible protective mechanism.METHODS:Sixty SD rats were randomly divided into four groups,15 rats in each group:the control group,model group,low-dose SPJ IVa group,and high-dose SPJ IVa group.A rat model of ALI was established via intratracheal instillation of lipopolysaccharide(LPS,2 mg/kg).Rats in the low-and high-dose SPJ IVa groups were intraperitoneally injected with 15 and 45 mg/kg SPJ IVa,respectively,30 min after model-ing.Serum,bronchoalveolar lavage fluid(BALF),and lungs were collected 24 h after modeling.Pathomorphological changes in lung tissues were assessed using HE staining.The wet weight/dry weight ratio of lung tissues was measured us-ing the weighing method,whereas ELISA was used to measure the levels of interleukin-1β(IL-1β),IL-6,and tumor ne-crosis factor-α(TNF-α)in the serum and BALF.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione(GSH)were assessed using the kit method.Cell apoptosis in lung tissues was evaluated by immunohisto-chemical staining of cleaved caspase-3 and TUNEL.Western blot was used to measure the expression of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),nuclear factor-κB(NF-κB)p65,and Toll-like receptor 4(TLR4)in lung tissues.RESULTS:Compared with control group,the lung tissues of the model group were significantly damaged,and the lung injury scores(0.21±0.22 vs 2.98±0.46)and lung wet/dry weight ratios(3.09±0.41 vs 6.36±0.61)were significantly increased(P<0.01).Compared with model group,the lung injury scores(1.80±0.31 and 1.05±0.25 vs 2.98±0.46)and lung wet/dry weight ratios(5.25±0.44 and 3.89±0.35 vs 6.36±0.61)in low-and high-dose SPJ IVa groups were significantly reduced(P<0.01).The administration of LPS resulted in elevated levels of pro-inflammatory cy-tokines(IL-1β,IL-6,and TNF-α)as well as the oxidative marker MDA in both serum and BALF(P<0.01).Additional-ly,it led to a decrease in antioxidant markers SOD and GSH(P<0.01).However,treatment with both low and high doses of SPJ IVa effectively attenuated the LPS-induced production of pro-inflammatory factors and oxidative markers MDA(P<0.01),while also increasing SOD and GSH levels(P<0.05 or P<0.01).In the model group,evident apoptosis was ob-served in lung tissues,whereas treatment with low and high doses of SPJ IVa significantly suppressed TUNEL-positive cells and the expression of cleaved caspase-3(P<0.01).The expression levels of Nrf2,HO-1,NF-κB p65,and TLR4 in lung tissues were significantly higher in the model group than in the control group(P<0.01);in turn,after treatment with low and high doses of SPJ IVa,Nrf2 and HO-1 were further upregulated(P<0.01),whereas NF-κB p65 and TLR4 were downregulated(P<0.01).CONCLUSION:The inhibitory effect of SPJ IVa on LPS-induced ALI in rats may be attribut-ed to its ability to suppress the TLR4/NF-κB-and Nrf2/HO-1-mediated inflammatory response and oxidative stress.

WIN55212-2 inhibits glycolysis and attenuates acute lung injury in septic mice by regulating mTOR/HIF-1α/PFKFB3 signaling pathway / 中国病理生理杂志

AIM:To investigate the effects of cannabinoid receptor agonist WIN55212-2(WIN)on acute lung injury(ALI)in septic mice,and to explore its potential mechanisms through glycolysis.METHODS:A mouse model of septic ALI was established by intraperitoneal injections of lipopolysaccharide(LPS).Male C57BL/6J mice were randomly divided into 4 groups(n=6):(1)control group;(2)LPS group,receiving intraperitoneal injections of LPS at 10 mg/kg;(3)LPS+WIN group,receiving 1 mg/kg WIN intraperitoneally 30 min prior to LPS injection;(4)LPS+WIN+MHY1485[mammalian target of rapamycin(mTOR)activator]group,receiving 10 mg/kg MHY1485 intraperitoneally 1 d before LPS injection and 1 mg/kg WIN plus 10 mg/kg MHY1485 30 min before LPS injection.Tissues were collected 24 h after modeling for analysis.Lung indexes were calculated,and histopathological changes of lung tissues were observed via he-matoxylin-eosin(HE)staining.Inflammatory cytokines interleukin-1β(IL-1β)and IL-10 in lung tissues,and lactic acid and lactate dehydrogenase A(LDHA)in serum were quantified using ELISA.The levels of mTOR/hypoxia-inducible fac-tor-1α(HIF-1α)/6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3)signaling pathway-related proteins were assessed by Western blot.RESULTS:Compared with control group,the LPS group exhibited an increased lung in-dex,significant lung tissue damage,an increase in IL-1β levels(P＜0.05),a decrease in IL-10 levels(P＜0.05),and el-evated expressions of lactate and LDHA(P＜0.05),along with increased levels of phosphorylated mTOR(p-mTOR),HIF-1α and PFKFB3 proteins(P＜0.05).The LPS+WIN group showed improvements with a reduced lung index(P＜0.05),lessened lung injury,decreased IL-1β levels(P＜0.05),increased IL-10 levels(P＜0.05),and lower levels of lactic acid,LDHA,p-mTOR,HIF-1α,and PFKFB3(P＜0.05).Conversely,the LPS+WIN+MHY1485 group displayed increased lung indexes and lung tissue damage,elevated IL-1β levels(P＜0.05),reduced IL-10 levels(P＜0.05),and higher expressions of lactic acid,LDHA,p-mTOR,HIF-1α and PFKFB3(P＜0.05)compared to the LPS+WIN group.CONCLUSION:WIN55212-2 mitigates sepsis-induced ALI,potentially by modulating the mTOR/HIF-1α/PFKFB3 sig-naling pathway,thereby inhibiting glycolysis and alleviating inflammatory responses.