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UTI attenuates HS-induced FITC-albumin extravasation in rat mesenteric postcapillary venules. The images of mesenteric postcapillary venules are shown. The FITC-albumin extravasation into the extravascular space is observed after HS, whereas UTI treatment prevented the extravasation.
Source publication
Recent studies have suggested that intrinsic apoptotic signaling cascade is involved in endothelial barrier dysfunction following hemorrhagic shock (HS), which results in vascular hyperpermeability. Our previous study demonstrated that ulinastatin (UTI) inhibits oxidant-induced endothelial hyperpermeability and apoptotic signaling. In present study...
Citations
... 40 41 It also appeared mitochondrial ROS were inhibited, a key mediator of apoptotic signalling. 40 Several therapies act on downstream processes from the intrinsic apoptotic signalling cascade, such an inhibition of smac release from the mitochondria. In a rat study, angiopoietin-1, known for inhibiting vascular permeability, attenuated HS-induced release of cytochrome c, smac and caspase-3 activity. ...
Introduction
Traumatic injury is one of the leading causes of death worldwide, and despite significant improvements in patient care, survival in the most severely injured patients remains unchanged. There is a crucial need for innovative approaches to improve trauma patient outcomes; this is particularly pertinent in remote or austere environments with prolonged evacuation times to definitive care. Studies suggest that maintenance of cellular homeostasis is a critical component of optimal trauma patient management, and as the cell powerhouse, it is likely that mitochondria play a pivotal role. As a result, therapies that optimise mitochondrial function could be an important future target for the treatment of critically ill trauma patients.
Methods
A systematic review of the literature was undertaken in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol to determine the potential role of mitochondria in traumatic injury and haemorrhagic shock (HS) and to identify current evidence for mitochondrial optimisation therapies in trauma. Articles were included if they assessed a mitochondrial targeted therapy in comparison to a control group, used a model of traumatic injury and HS and reported a method to assess mitochondrial function.
Results
The search returned 918 articles with 37 relevant studies relating to mitochondrial optimisation identified. Included studies exploring a range of therapies with potential utility in traumatic injury and HS. Therapies were categorised into the key mitochondrial pathways impacted following traumatic injury and HS: ATP levels, cell death, oxidative stress and reactive oxygen species.
Conclusion
This systematic review provides an overview of the key cellular functions of the mitochondria following traumatic injury and HS and identifies why mitochondrial optimisation could be a viable and valuable target in optimising outcome in severely injured patients in the future.
... In our previous studies, we reported the important role of the mitochondria-dependent apoptotic signaling pathway in VH [1,16]. Mitochondrial dysfunction, characterized by MMP depolarization and mitochondrial permeability transition pore opening, causes the release of second mitochondriaderived activator of caspase and cytochrome C into cytoplasm, which activates caspase-3. ...
... Then, cleavage of β-catenin by the activated caspase-3 regulates cell-cell adhesion in endothelial cells mediated by VE-cadherin, ultimately causing microvascular hyperpermeability. Therefore, maintaining mitochondrial homeostasis is crucial to the therapy for VH [4,16]. Mounting evidence highlights that the dynamics of mitochondrial fusion/fission are important in mitochondrial homeostasis [17,18]. ...
Mitochondria-dependent apoptotic signaling has a critical role in the pathogenesis of vascular hyperpermeability (VH). Dynamin-related protein-1- (Drp-1-) mediated mitochondrial fission plays an important role in mitochondrial homeostasis. In the present study, we studied the involvement of Drp-1 in resistance to VH induced by lipopolysaccharide (LPS). To establish the model of LPS-induced VH, LPS at 15 mg/kg was injected into rats in vivo and rat pulmonary microvascular endothelial cells were exposed to 500 ng/ml LPS in vitro. We found that depletion of Drp-1 remarkedly exacerbated the mitochondria-dependent apoptosis induced by LPS, as evidenced by reduced apoptosis, mitochondrial membrane potential (MMP) depolarization, and activation of caspase-3 and caspase-9. Increased FITC-dextran flux indicated endothelial barrier disruption. In addition, overexpression of Drp-1 prevented LPS-induced endothelial hyperpermeability and upregulated mitophagy, as evidenced by the loss of mitochondrial mass and increased PINK1 expression and mitochondrial Parkin. However, the mitophagy inhibitor, 3-Methyladenine, blocked these protective effects of Drp-1. Furthermore, inhibition of Drp-1 using mitochondrial division inhibitor 1 markedly inhibited LPS-induced mitophagy and aggravated LPS-induced VH, as shown by increased FITC-dextran extravasation. These findings implied that Drp-1 strengthens resistance to mitochondria-dependent apoptosis by regulating mitophagy, suggesting Drp-1 as a possible therapeutic target in LPS-induced VH.
... [20][21][22][23][24][25] Endothelial junctional proteins and cytoskeleton dynamics have a well-characterized role in endothelial health. 23,26,27 In addition, multiple reports utilizing shock models have shown that mitochondrial dysfunction contributes to increased microvascular permeability, [28][29][30] and we recently reported that disruption of mitochondrial complex III with antimycin A can severely impair endothelial barrier function. 29 As σ1 receptors reside in the ER-mitochondrial interface, 1 their ability to impact the endothelial barrier may be related to mitochondrial functions such as cellular bioenergetics. ...
... Because mitochondrial dysfunction has been reported to contribute to increased microvascular permeability in various models of shock, [28][29][30] we investigated whether PRE-084-induced enhancement of glycolytic ATP production could potentially serve as a barrier-protective mechanism. To accomplish this, we modeled mitochondrial dysfunction in cultured endothelial cell monolayers using the mitochondrial uncoupler, carbonyl cyanide m-chlorophenyl hydrazone (CCCP). ...
... Endothelial mitochondrial dysfunction has been reported in multiple models of microvascular hyperpermeability. [28][29][30] In the current study, we modeled this scenario using CCCP, which is known to cause mitochondrial damage, depolarization and energy depletion. 44,[47][48][49] We found that the σ1 agonist PRE-084 could partially reduce CCCP-induced endothelial barrier dysfunction, suggesting that σ1 could potentially serve as a useful therapeutic target to preserve endothelial barrier function in pathologies involving mitochondrial dysfunction. ...
Objective
We tested the hypothesis that sigma receptor‐1 (σ1) modulates endothelial barrier function due to its influence on endothelial bioenergetics.
Methods
Cultured human umbilical vein endothelial cell (HUVEC) monolayers were used to model the endothelial barrier. Electric cell‐substrate impedance sensing (ECIS), Transwell assays, and immunofluorescence labeling of junctional proteins were used to evaluate endothelial barrier function. Endothelial cell bioenergetics were determined using extracellular flux analysis and direct ATP level measurements. The endothelial‐specific contribution of σ1 was tested using the σ1‐selective agonist, PRE‐084, and with targeted knockdown of σ1 expression using siRNA.
Results
Activation of σ1 with PRE‐084 significantly enhanced endothelial barrier function and decreased permeability to albumin and dextran. Knockdown of σ1 with siRNA reduced barrier function and abolished PRE‐084‐induced endothelial barrier enhancement. PRE‐084 upregulated endothelial glycolysis and glycolytic ATP production, but this response was abolished by siRNA‐mediated knockdown of σ1 expression. PRE‐084 also reduced the degree of endothelial barrier dysfunction caused by the mitochondrial oxidative phosphorylation uncoupler CCCP.
Conclusion
Activation of σ1 enhances endothelial barrier function and modulates the ratio of glycolytic versus mitochondrial ATP production. These novel findings suggest that endothelial σ1 may prove beneficial as a novel therapeutic target for reducing microvascular hyperpermeability and counteracting mitochondrial dysfunction.
... ALI is a type of diffuse alveolar-capillary membrane injury characterized by pulmonary edema and atelectasis, and ARDS is a serious and life-threatening condition [1,2]. Studies showed that inflammation, mitochondrial damage, and apoptosis are involved in the etiology of ALI/ARDS [3]. Despite recent progress, the mortality rate associated with ALI/ARDS patients remains high [4]. ...
Mitophagy is involved in sepsis-induced acute lung injury (ALI). Bcl-2 family proteins play an important role in mitochondrial homeostasis. However, whether targeting Bcl-2 proteins (Bcl-2 and Bad) could influence mitophagy in ALI remains unclear. In this study, lipopolysaccharide (LPS) was used to induce injury in A549 cells and ALI in mice. LPS treatment resulted in elevated cell apoptosis, enhanced mitophagy, decreased Bcl-2 expression, increased Bad expression, and activation of PINK1/Parkin signaling in cells and lung tissues. Both Bcl-2 overexpression and Bad knockdown attenuated LPS-induced injury, inhibited cell apoptosis and mitophagy, and improved survival. Atg5 knockout (KO) inhibited LPS-induced cell apoptosis. Furthermore, Bcl-2 proteins regulated mitophagy by modulating the recruitment of Parkin from the cytoplasm to mitochondria via direct protein-protein interactions. These results were further confirmed in Park2 KO cells and Park2-/- mice. This is the first study to demonstrate that Bcl-2 proteins regulated mitophagy in LPS-induced ALI via modulating the PINK1/Parkin signaling pathway, promoting new insights into the mechanisms and investigation of therapeutic strategies for a septic patient with ALI.
... In present study, we hypothesised that PD mediates its protective effects against mitochondria-dependent apoptosis by upregulating mitophagy. Moreover, such mitochondria-dependent apoptosis is mediated by mitochondrial activation, resulting in the subsequent release of cytochrome c and second mitochondrial derived activator of caspases (smac), which ultimately leads to caspase-3 activation [7,30,31]. To investigate the role of Parkin-dependent mitophagy on PD-induced protection in ARDS, we assessed the effects of mitochondriadependent apoptosis while inhibiting mitophagy. ...
Mitophagy removes dysfunctional mitochondria and is known to play an important role in the pathogenesis of several diseases; however, the role of mitophagy in acute respiratory distress syndrome (ARDS) remains poorly understood. While we have previously demonstrated that polydatin (PD) improves lipopolysaccharide (LPS)-induced ARDS, the specific mechanism remains unclear. In present study, we aimed to determine whether PD activates Parkin-dependent mitophagy to protect against LPS-induced mitochondria-dependent apoptosis and lung injury. To establish the ARDS model, C57BL/6 mice were intratracheally injected with LPS (5 mg/kg) in vivo and Beas-2B cells were exposured to 0.5 mM LPS in vitro. Our results indicate that PD facilitates Parkin translocation to mitochondria and promotes mitophagy in ARDS-challenged mice and LPS-treated Beas-2B cells. However, PD-induced mitophagy was suppressed in Parkin−/− mice and Parkin siRNA transfected cells, indicating that PD activates Parkin-dependent mitophagy. Furthermore, the protective effects of PD against LPS-induced mitochondria-dependent apoptosis and lung injury were suppressed when Parkin was depleted both in vivo and in vitro. The inhibition of mitophagy with mitophagy inhibitor mitochondrial division inhibitor-1 in vivo and silencing of autophagy-related gene 7 in vitro also blocked the protective effects mediated by PD. Our data suggest that Parkin-dependent mitophagy induced by PD provides protection against mitochondria-dependent apoptosis in ARDS.
... Ulinastatin appears to attenuate ischemia-reperfusion injury in multiple organs by suppressing inflammation and oxidative stress [7,11,23], which reduces the generation of oxygen free radicals that accompanies many pathological states such as inflammation, ischemia, and reperfusion [12]. Ischemia-reperfusion injury can cause endothelial barrier dysfunction, resulting in high vascular permeability and tissue edema, and ulinastatin has been shown to inhibit vascular hyperpermeability [24,25]. Tissue edema reduces the supply of oxygen and nutrients, as well as the removal of waste products from tissues. ...
Background
Post-cardiac arrest syndrome, which has no specific curative treatment, contributes to the high mortality rate of victims who suffer traumatic cardiac arrest (TCA) and initially can be resuscitated. In the present study, we investigated the potential of ulinastatin to mitigate multiple organ injury after resuscitation in a swine TCA model.
Methods
Twenty-one male pigs were subjected to hemodynamic shock (40% estimated blood loss in 20 min) followed by cardiac arrest (electrically induced ventricular fibrillation) and respiratory suspension for 5 min, and finally manual resuscitation. At 5 min after resuscitation, pigs were randomized to receive 80,000 U/kg ulinastatin (n = 7) or the same volume of saline (n = 9) in the TCA group. Pigs in the sham group (n = 5) were not exposed to bleeding or cardiac arrest. At baseline and at 1, 3, and 6 h after the return of spontaneous circulation, blood samples were collected and assayed for tumor necrosis factor-alpha, interleukin 6, and other indicators of organ injury. At 24 h after resuscitation, pigs were sacrificed and apoptosis levels were assessed in samples of heart, brain, kidney, and intestine.
Results
One pig died in the ulinastatin group and one pig died in the TCA group; the remaining animals were included in the final analysis. TCA and resuscitation caused significant increases in multiple organ function biomarkers in serum, increases in tumor necrosis factor-alpha, and interleukin 6 in serum and increases in the extent of apoptosis in key organs. All these increases were lower in the ulinastatin group.
Conclusion
Ulinastatin may attenuate multiple organ injury after TCA, which should be explored in clinical studies.
Electronic supplementary material
The online version of this article (10.1186/s13017-018-0212-3) contains supplementary material, which is available to authorized users.
... [121][122][123][124] In animal trials, Ulinastatin suppressed the hyperpermeability of the vascular endothelium during BBB disruption. [129,132,135] Two studies suggest that UTI protects the integrity of the BBB by inhibiting the activation of caspase 3 and by preventing the disruption of adherens junctions. [130,132] Many novel interventions have failed because they are unable to function in the acidic environment created by lactic acidosis. ...
... [129,132,135] Two studies suggest that UTI protects the integrity of the BBB by inhibiting the activation of caspase 3 and by preventing the disruption of adherens junctions. [130,132] Many novel interventions have failed because they are unable to function in the acidic environment created by lactic acidosis. Ulinastatin is an acidic glycoprotein, and therefore acid-resistant which gives it the unique ability to effectively inhibit neurodegeneration during TBI despite acidic conditions. ...
... It has also been shown that Ulinastatin has significant antioxidant activity because it scavenges and clears away free radicals and it suppresses lipid peroxidation. [113,122,129,[131][132][133] of Ulinastatin also aids the preservation of mitochondrial function by preventing the collapse of mitochondrial membrane potential and by inhibiting the release of cytochrome c. [123,124,126,130,132] UTI further inhibits the formation of ROS from dysfunctional mitochondria. [132] By suppressing the generation of ROS and dysfunction of mitochondria, UTI indirectly inhibits intrinsic apoptosis. ...
... It has been suggested that the autophagic machinery plays an important role in regulating endothelial permeability 4,5 , but the mechanism of this regulation has not been clarified. In a previous study, we suggested that intrinsic apoptotic signaling (IAS) is involved in HS induced by VH 6 . In addition, autophagy exerts its protective effects by preventing apoptosis 7,8 . ...
... In a previous study, we reported that IAS plays an important role in VH 6,13 . In addition, GP has been demonstrated to ameliorate MD and apoptosis [14][15][16] . ...
... IAS is mediated by mitochondrial activation, resulting in the subsequent release of cytochrome c and second mitochondrialderived activator of caspases, and ultimately leading to caspase-3 activation 17,18 . Caspase-3 activation results in the cleavage of various cell adherens proteins, including β-catenin, which regulate VE-cadherin-mediated cell-cell adhesion in endothelial cells; 19 this cleavage may in turn lead to micro-VH 6,13 . In the present study, we observed activation of IAS following HS, as evidenced by MD, and indicated by collapsed MMP, lowered ATP levels, and increased cytoplasmic cytochrome c and caspase-3 activity. ...
Genipin (GP) is commonly used to treat cardiovascular diseases; however, the protective action of GP against vascular hyperpermeability (VH) has not been reported. We previously reported that intrinsic apoptotic signaling (IAS) is involved in VH following hemorrhagic shock (HS). GP inhibits apoptosis, but the specific mechanism remains unclear. In the present study, we observed that GP protects against HS-induced VH in vitro and in vivo. We report that this protective effect is related to the inhibition of IAS by up-regulation of autophagy via sirtuin 3 (SIRT3). The endothelial cell hyperpermeability induced by HS was enhanced by GP; this was attenuated by 3-methyladenine (3MA), a specific inhibitor of autophagy, indicating the involvement of autophagy. Consistent with these results, we found that 3MA reversed the effects of GP on up-regulation of autophagy, and also diminished the protective effect of GP against IAS activation following HS. Furthermore, knockout of SIRT3 inhibited GP-induced autophagy, indicating the requirement of SIRT3 in the regulation of autophagy by GP. In rats, GP improved HS-induced VH, which was repressed by 3MA and 3-(1H-1,2,3-triazol-4-yl)pyridine (3-TYP), a SIRT3 inhibitor. In conclusion, these findings suggest that autophagy plays a protective effect in VH following HS; the protective effect of autophagy is reinforced by GP, which protects against IAS and VH by up-regulating SIRT3.
... Ulinastatin's anti-inflammatory effects include inactivating the elastase secreted by neutrophils, decreasing inflammatory mediators and downregulating inflammatory transcription factors, including NF-κB (19,22,45). Under inflammatory conditions, ulinastatin can attenuate dysfunctions of the endothelial barrier by upregulating the expression of vascular endothelial-cadherin; it also prevents endothelial apoptosis (21,46,47). These effects against hyperpermeability may account for the protective effect of ulinastatin against heat-induced damage. ...
Intestinal injury-induced bacterial translocation and endotoxemia are important in the pathophysiological process of heatstroke. However, the underlying mechanism remains to be fully elucidated. Previous studies using 2D-gel electrophoresis found that defensin-related cryptdin-2 (Cry-2), an intestinal α-defensin, is upregulated in intestinal tissues during heatstroke in mice, and that treatment with ulinastatin, a multivalent enzyme inhibitor, reduced heat-induced acute lung injury. To investigate the association between Cry-2 and heat stress (HS)-induced intestinal injury and the probable protective role of ulinastatin, the present study examined the intestinal expression of Cry-2 via histopathologic analysis and reverse transcription-quantitative polymerase chain reaction analysis in mice with heatstroke. The heat-stressed mice were exposed to different core temperatures and cooling treatments, and intestinal pathological changes and Chiu scores were determined. Chemical markers of intestinal injury, serum and intestinal concentrations of diamine oxidase (DAO) and D-lactic acid (D-Lac), and serum and intestinal concentrations of Cry-2 were also determined. Correlations were analyzed using Spearman's correlation analysis. It was found that HS upregulated the expression of Cry-2, and the serum and intestinal concentrations of Cry-2 were correlated with the severity of HS-induced intestinal damage, indicated by pathology scores and concentrations of DAO and D-lac. Ulinastatin protected the intestines from HS-induced injury and downregulated the expression of Cry-2, which was also correlated with the extent of intestinal injury. Therefore, ulinastatin administration may be beneficial for patients with heatstroke, and Cry-2 may be a novel predictor of HS-induced intestinal injury.
... Moreover, Li et al (29) reported that ulinastatin inhibited cell hyperpermeability and apoptosis in oxidant-induced human umbilical endothelial cells. Lin et al (30) also revealed that ulinastatin attenuated the hyperpermeability of rat lung microvascular endothelial cells when treated by shock serum. All these results collaborate our results. ...
The present study aimed to evaluate the effects of ulinastatin on the permeability and apoptosis of lipopolysaccharide (LPS)-induced cardiac microvascular endothelial cells (CMVECs), and investigate its molecular mechanisms in sepsis. The sepsis rat model was induced by cecal ligation and puncture (CLP), and rat CMVECs were isolated and treated with LPS or/and ulinastatin. Then, cell permeability was evaluated by transendothelial electrical resistance, reactive oxygen species (ROS) levels were assessed by 2,7-dichlorofluorescein diacetate, cell apoptosis was detected using Annexin V-FITC apoptosis detection kit, and the expression levels of Bcl-2, Bax, caspase-3, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and EZH2 were detected by RT-qPCR and/or western blotting. In addition, the relationship of MALAT1 and EZH2 was evaluated by RNA immunoprecipitation (RIP) assay and chromatin immunoprecipitation (ChIP) assay. Compared with LPS-induced CMVECs, treatment with ulinastatin significantly reduced CMVEC permeability and the percentage of apoptotic cells, as well as an increased level of Bcl-2 and inhibited the levels of ROS, caspase-3 and Bax (all p<0.05). In addition, long non-coding RNA (lncRNA) MALAT1 was confirmed to interact with EZH2 in CMVECs. Overexpression of lncRNA MALAT1 and EZH2 was found in the hearts of the sepsis rat and LPS-induced CMVECs, while ulinastatin inhibited the upregulation of lncRNA MALAT1 and EZH2 in the LPS-induced CMVECs (all p<0.05). Ulinastatin protected against LPS-induced CMVEC cell hyperpermeability and apoptosis via downregulation of lncRNA MALAT1 and EZH2 in sepsis.
