Hypothermic machine perfusion (HMP) has been demonstrated to be more effective in mitigating ischemia–reperfusion injury (IRI) of donation after circulatory death (DCD) organs than cold storage (CS), yet the underlying mechanism remains obscure. We aimed to propose a novel therapeutic approach to ameliorate IRI in DCD liver transplantation. Twelve clinical liver samples were randomly assigned to HMP or CS treatment and subsequent transcriptomics analysis was performed. By combining in vivo HMP models, we discovered that HMP attenuated inflammation, oxidative stress, and apoptosis in DCD liver through a SEPRINA3-mediated PI3Kδ/AKT signaling cascade. Moreover, in the hypoxia/reoxygenation (H/R) model of BRL-3A, overexpression of SERPINA3 mitigated H/R-induced apoptosis, while SERPINA3 knockdown exacerbated cell injury. Idelalisib (IDE) treatment also reversed the protective effect of SERPINA3 overexpression. Overall, our research provided new insights into therapeutic strategies and identified potential novel molecular targets for therapeutic intervention against DCD liver.

Ischemia-reperfusion injury (IRI) is an inevitable pathological process during donation after circulatory death (DCD) liver transplantation, which contributes to serious damage to the graft. Oxidative stress, inflammation and apoptosis are all fatal causes of IRI of the liver. Hypothermic oxygenated perfusion (HOPE), as an emerging dynamic preservation technology, has a more significant effect on reducing DCD liver IRI than static cold storage (CS) mainly by regulating oxidative stress and inflammation. To further enhance the effect of HOPE and reveal its underlying mechanisms, investigators have recently combined HOPE with various methods. Excessive activation of the TLR/MyD88 signaling pathway can lead to severe immune inflammatory response. TJ-M2010-5 (TJ-5), a novel thiazaol-aminoramification MyD88 inhibitor, plays an essential role in the treatment of various diseases or pathological injuries in mice, such as hepatocellular carcinoma, acute liver injury and myocardial IRI. However, little is known about the role of TJ-5 in HOPE alleviating DCD liver IRI. Herein, we sought to investigate the role of HOPE combined with TJ-5 in reducing DCD liver IRI. We found that HOPE combined with TJ-5 significantly reduced oxidative stress, lessened inflammation, and decreased apoptosis during DCD liver IRI. Furthermore, HOPE combined with TJ-5 exerted their effects by inhibiting the TLR/MyD88 signaling pathway. Overall, these results demonstrated that HOPE combined with TJ-5 has a significant effect on alleviating DCD liver IRI. Therefore, the combined application of HOPE and TJ-5 may be an available and valid treatment option for DCD liver IRI.

血管麻痹综合征（vasoplegic syndromes, VS）是一种高排低阻的循环系统综合征,类似于热休克[1]。VS多见于大型手术之后,发生率达25%,其中以心血管外科手术和肝移植为最[2- 5]。相对于其他类型的手术患者,肝移植受者发生VS的风险更高[6]。一旦出现VS后诊治不及时,低血压和血液循环不稳定会严重影响移植物功能[7- 8]。目前,VS的治疗方法相对单一,缺乏标准方案[4- 5,9]。

Ischemia-reperfusion injury (IRI) is an inevitable and serious clinical problem in donations after heart death (DCD) liver transplantation. Excessive sterile inflammation plays a fateful role in liver IRI. Hypothermic oxygenated perfusion (HOPE), as an emerging organ preservation technology, has a better preservation effect than cold storage (CS) for reducing liver IRI, in which regulating inflammation is one of the main mechanisms. HECTD3, a new E3 ubiquitin ligase, and TRAF3 have an essential role in inflammation. However, little is known about HECTD3 and TRAF3 in HOPE-regulated liver IRI. Here, we aimed to investigate the effects of HOPE on liver IRI in a DCD rat model and explore the roles of HECTD3 and TRAF3 in its pathogenesis. We found that HOPE significantly improved liver damage, including hepatocyte and liver sinusoidal endothelial cell injury, and reduced DCD liver inflammation. Mechanistically, both the DOC and HECT domains of HECTD3 directly interacted with TRAF3, and the catalytic Cys (C832) in the HECT domain promoted the K63-linked polyubiquitination of TRAF3 at Lys138. Further, the ubiquitinated TRAF3 at Lys138 increased oxidative stress and activated the NF-κB inflammation pathway to induce liver IRI in BRL-3A cells under hypoxia/reoxygenation conditions. Finally, we confirmed that the expression of HECTD3 and TRAF3 was obviously increased in human DCD liver transplantation specimens. Overall, these findings demonstrated that HOPE can protect against DCD liver transplantation-induced-liver IRI by reducing inflammation via HECTD3-mediated TRAF3 K63-linked polyubiquitination. Therefore, HOPE regulating the HECTD3/TRAF3 pathway is a novel target for improving IRI in DCD liver transplantation.

Objective: To establish an animal model of experimental autoimmune myocarditis in Lewis rats and to investigate the expression and its significance of APRIL,a member of TNFSFs,in the experimental autoimmune myocarditis.Methods: Genetically predisposed Lewis rats were immunized with purified myosin plus complete Freund's adjuvant(CFA) in equal volume on day 1 and day 8,and the control rats were immunized with CFA only.The indexes of heart function(EF%,FS%,LVEDd and LVEDs) in the model group and control group were detected by echocardiogram on day 21 and day 90 after immunization.Hearts were harvested in both groups after the detection of echocardiogram.Histopathological changes of myocardium were measured by HE staining.The expression of APRIL mRNA was measured by reverse transcription polymerase chain reaction(RT-PCR).Results: On day 21 after immunization,inflammation was detected in the myocardium of experimental rats and inflammatory cells were mainly distributed around the blood vessels.On day 90,inflammatory cells decreased obviously and the myocardial cell became fibrosis,but no obvious inflammation emerged in the myocardium of control rats.The heart function in model group was worsen than in the control group(P0.05),but there was no significant difference between the two model groups(P0.05).The expression of APRIL mRNA was detected in the myocardium of experimental autoimmune myocarditis much more remarkably than in the normal myocardium(P0.05).Conclusion: The expression of APRIL mRNA increases in the myocardium of the experimental autoimmune myocarditis,and maybe play a role in the occurrence and development of autoimmune myocarditis.

Tumor necrosis factor-alpha (TNF-α), a proinflammatory cytokine involved in mitogen-activated protein kinase (MAPK) signaling pathways, contributes to the pathogenesis of cardiovascular diseases. Recently, suppressor of cytokine signaling-1 (SOCS-1) has been shown to modulate responses to TNF-α. However, whether SOCS-1 suppresses TNF-α-dependent apoptotic processes in cardiomyocytes and whether MAPK pathways mediate this effect have not been clearly elucidated. This study was carried out to define the role of SOCS-1 on TNF-α-induced apoptosis in neonatal rat cardiomyocytes and to investigate the signal pathways involved. Exposure to TNF-α (10 ng/ml for 24 h) significantly increased the number of apoptotic cells, the activity of caspase-8 and caspase-3, and the Bax/Bcl-xl ratio. In contrast, adenovirus-mediated gene transfer of SOCS-1 reversed the pro-apoptotic effect of TNF-α. Additionally, preincubation of cardiomyocytes with the extracellular signal-regulated kinase-1 and -2 (ERK1/2) inhibitor PD98059 attenuated the protective effect of SOCS-1, but the p38-MAPK inhibitor SB203580 and the c-Jun amino-terminal kinase (JNK) inhibitor SP600125 had no effect. Furthermore, the TNF-α-induced decrease in the phosphorylation of ERK1/2 was abolished by overexpression of SOCS-1. These findings suggest that SOCS-1 prevents TNF-α-induced apoptosis in cardiac myocytes via ERK1/2 pathway activation.

Objective: To establish animal model of experimental autoimmune myocarditis in Lewis rats and investigate the expression of LIGHT (TNFSF14) and HVEM in the experimental autoimmune myocarditis and its significance. Methods: Cardiac myosin was extracted from porcine ventricular myocardium. Genetically predisposed Lewis rats were immunized with cardiac myosin covered by complete Freund's adjuvant (CFA) on day 0 and 7 to establish the experiment autoimmune myocarditis(EAM) models. The control rats were immunized with CFA only. On day 21 and 90 after the first immunization, echocardiography was examined, histopathological changes of myocardium were detected by HE pigmentation, and gene levels of LIGHT and HVEM was measured by semi-quantitative RT-PCR. Results: In model rats, histopathological examination of cardiac tissue showed an obvious inflammatory cell infiltration with myocytes necrosis on day 21, simultaneously, echocardiography examination showed that heart function decreased as compared with control group(P<0.05). On day 90, fibrosis and a few of inflammatory cells could be observed. No inflammation and fibrosis was emerged in the myocardium of control rats. In addition, as compared with that in control groups, mRNA abundance for both LIGHT and HVEM were up-regulated (P<0.05) at acute phase (day 21) in the EAM models. On day 90, the mRNA abundance for LIGHT and HVEM was still up-regulated (P<0.05). Conclusion: The expression of LIGHT and HVEM mRNA in myocardium was up-regulated in experimental autoimmune myocarditis. The LIGHT/HVEM pathway maybe plays a role in the occurrence and development of autoimmune myocarditis.