Introduction: Dexamethasone (DEX), as an important enduring-effect glucocorticoid (GC), holds great promise in the field of lung ischemia-reperfusion injury (LIRI) comprehensive therapy owing to its immunomodulatory properties, such as inducing apoptosis and cell cycle distribution. However, its potent anti-inflammatory application is still restricted because of multiple internal physiologic barriers. Methods: Herein, we developed upconversion nanoparticles (UCNPs) coated with photosensitizer/capping agent/fluorescent probe-modified mesoporous silica (UCNPs@mSiO2[DEX]-Py/β-CD/FITC, USDPFs) for precise DEX release synergistic LIRI comprehensive therapy. The UCNPs were designed by covering an inert YOF:Yb shell on the YOF:Yb, Tm core to achieve high-intensity blue and red upconversion emission upon Near-Infrared (NIR) laser irradiation. Results: Under suitable compatibility conditions, the molecular structure of photosensitizer can be damaged along with capping agent shedding, which endowed USDPFs with an outstanding capability to carry out DEX release controlling and fluorescent indicator targeting. Furthermore, the hybrid encapsulating of DEX significantly increased utilization of nano-drugs, improving the water solubility and bioavailability, which was conducive to developing the anti-inflammatory performance of USDPFs in the complex clinical environment. Discussion: The response-controlled release of DEX in the intrapulmonary microenvironment can reduce normal cell damage, which can effectively avoid the side effects of nano-drugs in anti-inflammatory application. Meanwhile, the multi-wavelength of UCNPs endowed nano-drugs with the fluorescence emission imaging capacity in an intrapulmonary microenvironment, providing precise guidance for LIRI.

目的:基于秦皮素对铁死亡的调节作用,进一步探讨秦皮素对动脉粥样硬化(AS)的作用机制.方法:通过高脂饮食构建ApoE敲除(ApoE-/-)小鼠动脉粥样硬化模型,随机分为正常饮食组(NC组)、高脂饮食组(HFD组)、铁死亡抑制剂组(Fer-1组)、秦皮素组(Fra组)和萝卜硫素组(Sul组);用过氧化氢(H2O2)干预人原代脐静脉内皮细胞(HUVECs)构建体外AS模型,随机分为NC组、H2O2组、Fer-1组、Fra组和Sul组.油红O染色用于评估AS病变大小,血清低密度脂蛋白胆固醇(LDL-C)、血清高密度脂蛋白胆固醇(HDL-C)和总胆固醇(TC)测定试剂盒用于评估脂质代谢.细胞计数实验(CCK-8)用于检测秦皮素毒性作用和细胞增殖能力,铁、谷胱甘肽(GSH)和丙二醛(MDA)试剂盒用于测定小鼠主动脉组织和HUVECs中铁死亡水平.实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白免疫印迹(western blotting)实验主要通过检测谷胱甘肽过氧化物酶4(GPX4)和血红素加氧酶1(HMOX1)表达水平以评估秦皮素的抗铁死亡作用.结果:秦皮素能够减轻AS小鼠血清脂质沉积和主动脉粥样硬化,并且保护HUVECs免受H2O2诱导的细胞死亡.与Fer-1组结果一致,秦皮素在体内和体外均能抑制促铁死亡生物标志物铁和MDA表达水平,上调抗铁死亡生物标志物GSH和GPX4表达水平.此外,秦皮素减轻了H2O2诱导的HUVECs发生铁死亡,而这种保护作用能被HMOX1特异性激活剂萝卜硫素所逆转.结论:秦皮素可能通过下调HMOX1抑制铁死亡,从而减轻AS.

Abstract Background: Rheumatic heart disease (RHD) is an autoimmune disease caused by rheumatic fever following infection with Group A Streptococcus (GAS) that primarily affects the mitral valve, and our previous study has shown that endothelial-mesenchymal transition (EndMT) plays an important role in mitral valve damage due to RHD. TGF-β1 is an important profibrotic factor, but its role has not been explored in RHD. The current study is aimed to investigate the role of TGF-β1 in mitral valve damage due to RHD. Methods: RHD rats were established using inactivated GAS. Successful modelling of the RHD rat was verified in mitral valve pathology sections. Echocardiography and running test were used to detect the cardiac function of rats. And immunohistochemistry, RT-qPCR and Western blotting were used to detect activation of the TGF-β1/Smad signalling pathway and EndMT in mitral valve. Wounding healing and cell counting Kit-8 assay were conducted to figure out the function of TGF-β1 in vitro. Results: The TGF-β1/Smad signalling pathway was activated, and significant EndMT of the mitral valve occurred in RHD rats. And TGF-β type I receptor (TGF-βRI) inhibitor SB431542 inhibited the TGF-β1/Smad signalling pathway and EndMT of the mitral valve. Conclusion: EndMT was involved in mitral valve damage, and inhibition of TGF-βRI alleviated EndMT of mitral valve damage due to RHD.

Lung ischemia-reperfusion injury (LIRI) is a complex "aseptic" inflammatory response, macrophage play a pivotal role in the pathogenesis of LIRI. Galectin-3 (Gal3), a lectin implicated inflammation, has received limited attention in LIRI. Studies have reported Gal3 as a ligand for triggering receptor expressed on myeloid cell 2 (TREM2) in macrophages in Alzheimer's disease. Hence, we established LIRI C57BL/6 mice model and hypoxia/glucose deprivation and reoxygenation (OGD/R) model to investigate the relationship among Gal3, TREM2, and macrophage polarization. Our result demonstrated inhibition of Gal3 significantly reduced M1-type macrophage polarization while markedly increased M2-type in LIRI. In addition, we observed colocalization of Gal3 and TREM2 in macrophages, inhibition of Gal3 could recover the downregulation of TREM2 induced by LIRI while promoting TREM2 expression could attenuate lung injury in LIRI. In summary, our findings suggest Gal3 as an upstream factor of TREM2, play a crucial role in LIRI by regulating macrophage polarization.

Lung ischemia/reperfusion injury (LIRI) is a complex pathophysiological process, with the histopathological hallmark of neutrophils migrating into the lungs. Neutrophil extracellular traps (NETs) have been suggested to exert a critical role in the pathogenesis of inflammation and infection in humans and animals, while the exact functions and underlying mechanisms of NETs in LIRI remain insufficiently elucidated. In this study, we investigated the role of pore-forming protein gasdermin D (GSDMD) on NETs release in LIRI induced by lung ischemia/reperfusion (I/R). We found that disulfiram, a GSDMD inhibitor, dramatically reduced NETs release and pathological injury in lung I/R in vivo and in vitro. Additionally, GSDMD caused mitochondrial DNA (mtDNA) leaking into the neutrophil cytosol, and then the cytoplasmic mtDNA activated the cGAS-STING signaling pathway and stimulated NETs formation in lung I/R. Furthermore, inhibition of cGAS/STING pathway could inhibit cytosol mtDNA mediated NETs formation.

目的:探讨风湿性心脏病(RHD)大鼠的血清对内皮细胞发生内皮间充质转化(EndMT)的影响.方法:先使用重组人转化生长因子-β1(TGF-β1)干预人原代脐静脉内皮细胞(HUVECs)以观察其发生EndMT的情况,分为空白组和TGF-β1组;再使用大鼠血清和TGF-βⅠ型受体(TGF-βRⅠ)抑制剂SB431542干预HUVECs,分为control组、RHD组和RHD+SB431542组,采用细胞计数试剂盒(CCK-8)检测细胞增殖能力,采用细胞划痕实验检测细胞迁移能力,采用实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白免疫印迹(western blotting)实验检测内皮细胞标志分子血管内皮细胞钙粘连蛋白(VE-cadherin)和间质细胞标志分子α-平滑肌肌动蛋白(α-SMA)水平以观察细胞发生EndMT的情况.结果:与control组比较,RHD组的细胞增殖、迁移能力显著升高(P<0.05),且VE-cadherin蛋白的表达量降低,而α-SMA蛋白的表达量升高(P<0.05),表明发生了EndMT;而与RHD组比较,RHD+SB431542组的细胞增殖、迁移能力显著降低(P<0.05),且VE-cadherin蛋白的表达量升高,而α-SMA蛋白的表达量降低(P<0.05),表明EndMT被显著抑制.结论:RHD大鼠的血清高表达TGF-β1,可以诱导HUVECs发生End-MT,且能被SB431542抑制,表明在RHD大鼠血清促进内皮细胞间充质转化过程中,TGF-β1发挥着重要作用.

Abstract MicroRNA (miR)-143-3p is a potential regulatory molecule in myocardial ischemia/reperfusion injury (MI/RI), wherein its expression and pathological effects remains controversial. Thus, a mouse MI/RI and cell hypoxia/reoxygenation (H/R) models were built for clarifying the miR-143-3p’s role in MI/RI. Following myocardial ischemia for 30 min, mice underwent reperfusion for 3, 6, 12 and 24 h. It was found miR-143-3p increased in the ischemic heart tissue over time after reperfusion. Cardiomyocytes transfected with miR-143-3p were more susceptible to apoptosis. Mechanistically, miR-143-3p targeted B cell lymphoma 2 (bcl-2). And miR-143-3p inhibition reduced cardiomyocytes apoptosis upon H/R, whereas it was reversed by a specific bcl-2 inhibitor ABT-737. Of note, miR-143-3p inhibition upregulated bcl-2 with better mitochondrial membrane potential (Δψm), reduced cytoplasmic cytochrome c (cyto-c) and caspase proteins, and minimized infarction area in mice upon I/R. Collectively, inhibition of miR-143-3p might alleviate MI/RI via targeting bcl-2 to limit mitochondria-mediated apoptosis. To our knowledge, this study further clarifies the miR-143-3p’s pathological role in the early stages of MI/RI, and inhibiting miR-143-3p could be an effective treatment for ischemic myocardial disease.

目的:探讨生长转化因子15(GDF15)对过氧化氢(H2O2)诱导的内皮细胞损伤和血管生成的影响.方法:用H2O2诱导人脐静脉内皮细胞(HUVECs)氧化损伤模型,分为对照组、H2O2组、小分子干扰RNA(siRNA)靶向沉默GDF15(si-GDF15)组和si-GDF15+H2O2组.采用CCK-8检测细胞存活率,流式细胞术检测细胞活性氧(ROS)的浓度和凋亡率,划痕实验检测细胞迁移能力,小管形成实验检测内皮细胞成管能力.结果:与对照组比较,H2O2组细胞ROS浓度和凋亡率显著增高,细胞增殖、迁移和小管生成能力显著降低(均P＜0.05);与H2O2组比较,si-GDF15+H2O2组细胞ROS和凋亡率显著增高,细胞增殖、迁移和小管生成能力显著降低(均P＜0.05).结论:沉默GDF15能增强H2O2诱导的内皮损伤,并抑制HUVECs的血管形成能力.

Acute lung injury (ALI) is a common clinically critical illness that can develop into acute respiratory distress syndrome and even lead to death. Currently, there is no effective treatment method. In different pathophysiological conditions, neutrophils manifest many different forms of death, such as neutrophil extracellular traps, ferroptosis, pyroptosis, apoptosis, autophagy, necrosis and necroptosis. Different forms of neutrophil death play distinct roles in the occurrence and development of ALI. This review summarizes the different forms of neutrophil death and their role in ALI and aims to provide new ideas and new targets for exploring the mechanism and prevention and treatment strategies of ALI.

目的:探讨半乳凝集素3(Ga13)在小鼠肺缺血再灌注损伤(LIRI)模型中的表达情况.方法:将20只C57BL/6J雄性小鼠随机分为假手术组和4个实验组(即LIRI后2 h组、LIRI后6 h组、LIRI后12 h组、LIRI后24 h组),每组4只.假手术组只开胸不夹闭肺门,实验组夹闭肺门60min并分别于松开血管夹后2h、6h、12h、24h收集左肺标本.采用苏木精—伊红(HE)染色法和肺组织湿干重比(W/D)观察肺组织病理学变化,酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液(BALF)炎性因子白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-a)的表达,分别采用实时荧光定量PCR(RT-qPCR)、western blotting、免疫荧光染色法检测Gal3 mRNA与Ga13蛋白的表达水平.结果:与假手术组相比,LIRI后各实验组出现肺组织损伤,组织结构破坏明显,肺损伤评分和W/D均升高,炎症因子IL-1β和TNF-a水平升高(均P＜0.05),在LIRI后6h肺组织病理学损伤程度和炎性因子水平达到峰值(P＜0.05),随后逐渐降低.与假手术组相比,LIRI后各实验组Ga13 mRNA及其蛋白表达水平升高,Ga13免疫荧光染色表达水平升高(P＜0.05),均于LIRI后6 h达到高峰(P＜0.05),然后逐渐下降.结论:小鼠LIRI后Ga13表达升高,高水平的Ga13可能与LIRI后的肺组织损伤和炎症爆发有关.

Colorectal cancer (CRC) is currently one of the commonest tumors and the main reason for cancer-related deaths worldwide. It has been reported that long non-coding RNAs (lncRNAs) act as important indicators and regulators in various cancers. There is an urgent need to explore new lncRNA biomarkers in CRC, as well as their functions and molecular mechanisms. NALT1 has been implicated in the occurrence of gastric cancer (GC). However, the detailed function and mechanism of NALT1 in CRC progress have not been reported. In this study, RT-qPCR was conducted to detect the expression of NALT1 in 76 CRC patients ranging from stages I through IV. To assess the biological function of NALT1, loss- and gain-of-function experiments were conducted both in vivo and in vitro. Moreover, RNA-seq, bioinformatics analysis, RNA pulldown assay, dual-luciferase reporter, Ago2-RIP, quantitative PCR, Western blot assays, and rescue experiments were performed to reveal the molecular mechanisms of competitive endogenous RNAs (ceRNAs). It was observed that high expression of NALT1 was markedly correlated with advanced cancer stage in the clinic. Functionally, NALT1 downregulation inhibited cell proliferation, migration and invasion, whereas NALT1 overexpression exhibited an opposite trend both in vivo and in vitro. Bioinformatics analysis, RNA pulldown, Ago2-RIP, and luciferase reporter assays showed that miRNA-574-5p was a target of NALT1. Additionally, dual-luciferase reporter assays, Ago2-RIP, and rescue experiments indicated that miRNA-574-5p could target the PEG10 gene directly. Our results suggested that NALT1 promoted CRC proliferation and migration by sponging miRNA-574-5p to upregulate PEG10 expression, and implied that NALT1 might act as a promising biomarker and therapeutic target for CRC.

Lung ischemia-reperfusion injury (LIRI) is a severe multifaceted pathological condition that can lead to poor patient outcome where oxidative stress and the resulting inflammatory response can trigger and exacerbate tissue damage in LIRI patients. Sirtuin3 (SIRT3), a member of the sirtuin family, protects against oxidative stress-related diseases. However, it remains unclear if and how SIRT3 alleviates lung injury induced by ischemia/reperfusion (I/R). Our previous study showed that lung tissue structures were severely damaged at 6 h after lung I/R in mice, however, repair of the injured lung tissue was significant at 24 h. In this study, we found that both SIRT3 mRNA and protein levels were markedly increased at 24 h after lung I/R in vivo. Meanwhile, inhibition of SIRT3 aggravated lung injury and inflammation, augmented mitochondrial fission and oxidative stress and increased Hypoxia-inducible factor-1α (HIF-1α) expression in vivo. The results suggest that SIRT3 may be an upstream regulator of HIF-1α expression. Knockdown of SIRT3 resulted in excessive mitochondrial fission and increased oxidative stress in vitro, and we found that knocking down the expression of HIF-1α alleviated these changes. This suggests that the SIRT3-HIF-1α signaling pathway is involved in regulating mitochondrial function and oxidative stress. Furthermore, inhibition of dynamin-related protein 1 (Drp-1) by the inhibitor of mitophagy, Mdivi-1, blocked mitochondrial fission and alleviated oxidative stress in vitro. Taken together, our results demonstrated that downregulation of SIRT3 aggravates LIRI by increasing mitochondrial fission and oxidative stress. Activation of SIRT3 inhibits mitochondrial fission and this mechanism may serve as a new therapeutic strategy to treat LIRI.

Lung ischemia reperfusion injury (LIRI) is a complex pathophysiological process with high morbidity and mortality. An important pathophysiological characteristic of LIRI is endothelial barrier dysfunction, although the mechanism involved in this process remains unclear. VX765, a specific caspase-1 inhibitor, has been shown to have a protective effect against several diseases including sepsis, atherosclerosis, and glial inflammatory disease. The objective of this study was to determine whether VX765 had a protective effect in LIRI. The results showed that lung ischemia/reperfusion (I/R) and oxygen/glucose deprivation and reoxygenation (OGD/R) induced endothelial pyroptosis and barrier dysfunction characterized by an inflammatory response. Treatment with VX765 successfully alleviated I/R- and OGD/R-induced endothelial pyroptosis and barrier dysfunction by inhibiting caspase-1 in vivo and in vitro. In conclusion, these findings showed that VX765 provided effective protection against lung I/R-induced endothelial pyroptosis and barrier dysfunction.

目的:探讨热休克蛋白60(HSP60)调控2型髓系细胞触发受体(TREM2)对肺缺血再灌注损伤(LIRI)小鼠的保护作用.方法:将24只C57BL/6小鼠随机分成对照组、HSP60组、LIRI组、LIRI+HSP60组,每组6只.LIRI+HSP60组于术前1 h腹腔注射HSP60(5 μg/g).HSP60组腹腔注射等剂量的HSP60;对照组不做任何处理.LIRI组和LIRI+HSP60组小鼠建立LIRI模型,术后6 h收集左肺组织标本,苏木精—伊红(HE)染色法观察肺组织病理学变化,透视电镜观察肺组织超微结构改变,酶联免疫吸附法(ELISA)检测肺组织中炎性因子白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)的表达,实时荧光定量PCR(qPCR)法与Western blotting检测TREM2 mRNA及TREM2蛋白的相对表达水平.结果:与对照组和HSP60组比较,LIRI组肺组织损伤严重,肺组织结构改变明显,肺损伤评分明显增加(P＜0.05),炎症因子IL-1β和IL-18水平均明显升高(均P＜0.05),TREM2 mRNA及其蛋白相对表达水平均明显降低(均P＜0.05).与LIRI组比较,LIRI+HSP60组肺组织病理学损伤明显改善,肺损伤评分明显降低(P＜0.05),炎症因子IL-1β和IL-18表达明显减少(均PP＜0.05),TREMM2 mRNA及其蛋白的相对表达水平均明显升高(均P＜0.05).与对照组相比较,HSP60组TREM2mRNA及蛋白的相对表达水平均明显升高(P＜0.05).结论:HSP60可以减轻LIRI,其机制可能与上调TREM2的表达有关.

Objective:To investigate the role and regulatory mechanism of triggering receptor expressed on myeloid cell 2 (TREM2) in mice lung ischemia/reperfusion injury (LIRI).Methods:Thirty-six healthy male C57BL/6 mice were divided into six groups according to the random number method ( n = 6): normal control group, and LIRI 2, 6, 12, 24, 48 hours group. Mice LIRI models were established by clamping the left hilum. The wet/dry weight ratio (W/D) of left lung tissue was measured. Lung injury was observed and evaluated by hematoxylin-eosin (HE) staining and electron microscopy. The levels of interleukins (IL-1β, IL-18) in lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of TREM2 and caspase-1 were determined by polymerase chain reaction (PCR). The protein expressions of TREM2, caspase-1, Gasdermin-D (GSDMD) were determined by Western blotting. Results:At 2 hours after LIRI, lung injury began to appear, the lung ultrastructure changed, and the lung injury score increased; at 6 hours, the degree of lung injury was the most serious; after 12 hours, the lung injury gradually reduced and the lung injury score gradually decreased. Compared with the normal control group, lung W/D ratio and lung injury score of LIRI 2, 6, 12, 24, 48 hours groups were significantly higher, the differences were statistically significant (lung W/D ratio: 7.06±0.52, 8.34±0.17, 6.42±0.35, 5.34±0.25, 5.59±0.45 vs. 4.69±0.23; lung injury score: 5.50±0.54, 9.75±0.89, 5.88±0.84, 3.63±0.74, 4.13±0.64 vs. 1.13±0.35, all P < 0.05). Compared with the normal control group, the levels of IL-1β and IL-18 in lung tissue were significantly increased at 2 hours after LIRI, reached a peak at 6 hours [IL-1β (ng/L): 502.76±12.25 vs. 56.50±8.07, IL-18 (ng/L): 414.02±10.75 vs. 81.63±5.29, both P < 0.05], then decreased gradually, and were still significantly higher than the normal control group at 48 hours. The PCR and Western blotting showed that the expression of TREM2 was significantly lower than that in the normal control group at 2 hours after LIRI, and reached a valley at 6 hours [TREM2 mRNA (2 -ΔΔCt): 0.47±0.05 vs. 1.02±0.05, TREM2/GAPDH: 0.23±0.13 vs. 0.48±0.17, both P < 0.05], then gradually increased, and reached the peak at 24 hours [TREM2 mRNA (2 -ΔΔCt): 3.98±0.15 vs. 1.02±0.05, TREM2/GAPDH: 0.71±0.17 vs. 0.48±0.17, both P < 0.05]. The trend of expression of caspase-1 and GSDMD were opposite to that of TREM2, which increased at first and then decreased, and reached a peak at 6 hours after reperfusion [caspase-1 mRNA (2 -ΔΔCt): 2.20±0.13 vs. 1.01±0.02, caspase-1/GAPDH: 0.64±0.02 vs. 0.20±0.06, GSDMD/GAPDH: 1.23±0.01 vs. 0.87±0.02, all P < 0.05]. Conclusions:TREM2 might be involved in LIRI in mice. The mechanism may be related to the effect of TREM2 on caspase-1-mediated pyroptosis.

Background: Lung ischemia reperfusion injury (LIRI) is a complex pathophysiological process activated by lung transplantation and acute lung injury. The p38 mitogen-activated protein kinase (MAPK) is involved in breakdown of the endothelial barrier during LIRI, but the mechanism is still unclear. Therefore, we investigated the function of p38 MAPK in LIRI in vivo and in vitro . Methods: Sprague–Dawley rats were subjected to ischemia reperfusion with or without pretreatment with a p38 MAPK inhibitor. Lung injury was assessed using hematoxylin and eosin staining, and pulmonary blood–air barrier permeability was evaluated using Evans blue staining. A rat pulmonary microvascular endothelial cell line was infected with lentiviral expressing short hairpin (sh)RNA targeting p38 MAPK and then cells were subjected to oxygen/glucose deprivation and reoxygenation (OGD/R). Markers of endothelial destruction were measured by western blot and immunofluorescence. Results: In vivo LIRI models showed structural changes indicative of lung injury and hyperpermeability of the blood–air barrier. Inhibiting p38 MAPK mitigated these effects. Oxygen/glucose deprivation and reoxygenation promoted hyperpermeability of the endothelial barrier in vitro , but knockdown of p38 MAPK attenuated cell injury; maintained endothelial barrier integrity; and partially reversed injury-induced downregulation of permeability protein AQP1, endothelial protective protein eNOS, and junction proteins ZO-1 and VE-cadherin while downregulating ICAM-1, a protein involved in destroying the endothelial barrier, and ET-1, a protein involved in endothelial dysfunction. Conclusion: Inhibition of p38 MAPK alleviates LIRI by decreasing blood–air hyperpermeability. Blocking p38 MAPK may be an effective treatment against acute lung injury.