Ischemia‐reperfusion injury. A phenomenon,that also occurs in the bone

Ischemia‐reperfusion injury of the bone occurs due to traumatic and non‐traumatic alterations affecting blood supply to the bone. It is likely to occur also upon insertion of an implant. Ischemia‐reperfusion injury of the bone has been studied by interruption of blood supply in situ, in limb replantation/transplantation models, in revascularized bone grafts and non‐vascularized bone fragments, as well as in isolated cultured cells. All cells of the bone are affected, including osteoblasts, osteocytes, osteoclasts, chondrocytes, and bone marrow cells. Critical ischemia times for induction of bone cell death, either in the ischemic period or following reperfusion, are in the range of 3 to 7 h. These critical ischemia times are significantly increased by decreasing the temperature from 37 °C to 0–4 °C. Anoxia is the most likely trigger of cell injury in the ischemic phase. In the reperfusion phase, reactive oxygen species are decisively involved in the injurious process. In general, however, the available information on the mechanism of ischemia‐reperfusion injury of the bone is relatively sparse. On the other hand, there are clear similarities to the mechanisms of ischemia‐reperfusion injury known from other organs, and there is a clear potential for protection against ischemia‐reperfusion injury of the bone.     

P2Y11 Agonism Prevents Hypoxia/Reoxygenation- and Angiotensin II-Induced Vascular Dysfunction and Intimal Hyperplasia Development

Vascular dysfunction in cardiovascular diseases includes vasomotor response impairments, endothelial cells (ECs) activation, and smooth muscle cells (SMCs) proliferation and migration to the intima. This results in intimal hyperplasia and vessel failure. We previously reported that activation of the P2Y11 receptor (P2Y11R) in human dendritic cells, cardiofibroblasts and cardiomyocytes was protective against hypoxia/reoxygenation (HR) lesions. In this study, we investigated the role of P2Y11R signaling in vascular dysfunction. P2Y11R activity was modulated using its pharmacological agonist NF546 and antagonist NF340. Rat aortic rings were exposed to angiotensin II (AngII) and evaluated for their vasomotor response. The P2Y11R agonist NF546 reduced AngII-induced vascular dysfunction by promoting EC-dependent vasorelaxation, through an increased nitric oxide (NO) bioavailability and reduced AngII-induced H₂O₂ release; these effects were prevented by the use of the P2Y11R antagonist NF340. Human vascular SMCs and ECs were subjected to AngII or H/R simulation in vitro. P2Y11R agonist modulated vasoactive factors in human ECs, that is, endothelial nitric oxide synthase (eNOS) and endothelin-1, reduced SMC proliferation and prevented the switch towards a synthetic phenotype. H/R and AngII increased ECs secretome-induced SMC proliferation, an effect prevented by P2Y11R activation. Thus, our data suggest that P2Y11R activation may protect blood vessels from HR-/AngII-induced injury and reduce vascular dysfunctions. These results open the way for new vasculoprotective interventions.     

Ivabradine Induces Cardiac Protection against Myocardial Infarction by Preventing Cyclophilin-A Secretion in Pigs under Coronary Ischemia/Reperfusion

In response to cardiac ischemia/reperfusion, proteolysis mediated by extracellular matrix metalloproteinase inducer (EMMPRIN) and its secreted ligand cyclophilin-A (CyPA) significantly contributes to cardiac injury and necrosis. Here, we aimed to investigate if, in addition to the effect on the funny current (I(f)), Ivabradine may also play a role against cardiac necrosis by reducing EMMPRIN/CyPA-mediated cardiac inflammation. In a porcine model of cardiac ischemia/reperfusion (IR), we found that administration of 0.3 mg/kg Ivabradine significantly improved cardiac function and reduced cardiac necrosis by day 7 after IR, detecting a significant increase in cardiac CyPA in the necrotic compared to the risk areas, which was inversely correlated with the levels of circulating CyPA detected in plasma samples from the same subjects. In testing whether Ivabradine may regulate the levels of CyPA, no changes in tissue CyPA were found in healthy pigs treated with 0.3 mg/kg Ivabradine, but interestingly, when analyzing the complex EMMPRIN/CyPA, rather high glycosylated EMMPRIN, which is required for EMMPRIN-mediated matrix metalloproteinase (MMP) activation and increased CyPA bonding to low-glycosylated forms of EMMPRIN were detected by day 7 after IR in pigs treated with Ivabradine. To study the mechanism by which Ivabradine may prevent secretion of CyPA, we first found that Ivabradine was time-dependent in inhibiting co-localization of CyPA with the granule exocytosis marker vesicle-associated membrane protein 1 (VAMP1). However, Ivabradine had no effect on mRNA expression nor in the proteasome and lysosome degradation of CyPA. In conclusion, our results point toward CyPA, its ligand EMMPRIN, and the complex CyPA/EMMPRIN as important targets of Ivabradine in cardiac protection against IR.     

Cyclic Nigerosyl-Nigerose as Oxygen Nanocarrier to Protect Cellular Models from Hypoxia/Reoxygenation Injury: Implications from an In Vitro Model

Heart failure (HF) prevalence is increasing among the aging population, and the mortality rate remains unacceptably high despite improvements in therapy. Myocardial ischemia (MI) and, consequently, ischemia/reperfusion injury (IRI), are frequently the basis of HF development. Therefore, cardioprotective strategies to limit IRI are mandatory. Nanocarriers have been proposed as alternative therapy for cardiovascular disease. Controlled reoxygenation may be a promising strategy. Novel nanocarriers, such as cyclic nigerosyl-nigerose (CNN), can be innovative tools for oxygen delivery in a controlled manner. In this study we analyzed new CNN-based formulations as oxygen nanocarriers (O₂-CNN), and compared them with nitrogen CNN (N₂-CNN). These different CNN-based formulations were tested using two cellular models, namely, cardiomyoblasts (H9c2), and endothelial (HMEC) cell lines, at different concentrations. The effects on the growth curve during normoxia (21% O₂, 5% CO₂ and 74% N₂) and their protective effects during hypoxia (1% O₂, 5% CO₂ and 94% N₂) and reoxygenation (21% O₂, 5% CO₂ and 74% N₂) were studied. Neither O₂-CNN nor N₂-CNN has any effect on the growth curve during normoxia. However, O₂-CNN applied before hypoxia induces a 15–30% reduction in cell mortality after hypoxia/re-oxygenation when compared to N₂-CNN. O₂-CNN showed a marked efficacy in controlled oxygenation, which suggests an interesting potential for the future medical application of soluble nanocarrier systems for MI treatment.     

Hydrogen Sulfide Metabolite,Sodium Thiosulfate: Clinical Applications and Underlying Molecular Mechanisms

Thiosulfate in the form of sodium thiosulfate (STS) is a major oxidation product of hydrogen sulfide (H₂S), an endogenous signaling molecule and the third member of the gasotransmitter family. STS is currently used in the clinical treatment of acute cyanide poisoning, cisplatin toxicities in cancer therapy, and calciphylaxis in dialysis patients. Burgeoning evidence show that STS has antioxidant and anti-inflammatory properties, making it a potential therapeutic candidate molecule that can target multiple molecular pathways in various diseases and drug-induced toxicities. This review discusses the biochemical and molecular pathways in the generation of STS from H₂S, its clinical usefulness, and potential clinical applications, as well as the molecular mechanisms underlying these clinical applications and a future perspective in kidney transplantation.     

肾缺血再灌注细胞内钙水平与氧化应激损伤研究

目的 观察肾细胞中游离钙([Ca~(2+)]i)及氧自由基在缺血再灌注损伤过程中的改变情况,探讨二者在再灌注肾损伤中发生作用机制。方法 摘除Wistar大鼠左肾,钳夹右侧肾蒂,建立急性缺血再灌注肾损伤模型,应用Fura-2/AM荧光指示剂测定缺血再灌注大鼠肾细胞内[Ca~(2+)]i浓度的变化,同时测定谷胱甘肽过氧化物酶(GSH-Px)活性、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果 缺血再灌注不同时期肾细胞内[Ca~(2+)]i浓度均有不同程度增高,与对照组相比差异有显著意义,各实验组SOD活力水平降低,与对照组相比差异有显著意义,MDA生成高于对照组,GSH-Px在再灌注早期活力减弱,明显低于对照组,晚期活力基本恢复。结论 1.缺血再灌注不同时期大鼠肾细胞内[Ca~(2+)]i超载和不同程度的氧化侵袭,在缺血再灌注肾损伤病理过程中起重要作用;2.再灌注时间与肾细胞内[Ca~(2+)]i超载呈正相关,提示再灌注损伤通过不同机制加重细胞钙超载。     

Texture analysis of poly-adenylated mRNA staining following global brain ischemia and reperfusion

Texture analysis provides a means to quantify complex changes in microscope images. We previously showed that cytoplasmic poly-adenylated mRNAs form mRNA granules in post-ischemic neurons and that these granules correlated with protein synthesis inhibition and hence cell death. Here we utilized the texture analysis software MaZda to quantify mRNA granules in photomicrographs of the pyramidal cell layer of rat hippocampal region CA3 around 1 h of reperfusion after 10 min of normothermic global cerebral ischemia. At 1 h reperfusion, we observed variations in the texture of mRNA granules amongst samples that were readily quantified by texture analysis. Individual sample variation was consistent with the interpretation that animal-to-animal variations in mRNA granules reflected the time-course of mRNA granule formation. We also used texture analysis to quantify the effect of cycloheximide, given either before or after brain ischemia, on mRNA granules. If administered before ischemia, cycloheximide inhibited mRNA granule formation, but if administered after ischemia did not prevent mRNA granulation, indicating mRNA granule formation is dependent on dissociation of polysomes. We conclude that texture analysis is an effective means for quantifying the complex morphological changes induced in neurons by brain ischemia and reperfusion.     

丹参素对大鼠局灶性脑缺血再灌注损伤的保护作用

目的研究丹参素对大鼠局灶性脑缺血再灌注损伤的保护作用,并初步探讨其作用机制.方法用线栓法制备大鼠局灶性脑缺血再灌注损伤模型,通过神经功能学评分、苏木素和伊红(HE)染色及流式细胞术检测丹参素对大鼠脑缺血再灌注损伤后行为学、缺血侧脑组织病理形态学及神经细胞凋亡的影响.结果丹参素可明显改善脑缺血大鼠神经功能缺损体征,减轻脑组织病理形态学损伤,降低神经细胞凋亡百分率.结论丹参素对大鼠局灶性脑缺血再灌注损伤具有保护作用,其作用机制可能与抑制神经细胞凋亡有关.     

核桃中血清素的提取分离与抗心肌缺血活性研究

核桃破碎后经体积分数为70%乙醇提取,乙酸乙酯萃取、浓缩后,经大孔树脂、MCI、ODS等不同层析柱分离。采用MTT法和试剂盒法,以乳鼠心肌细胞缺血/再灌注损伤模型检测各分离组分抗心肌缺血活性。结果从核桃中分离得到5,10-二羟色胺和Shepherdine化合物,其中5,10-二羟色胺能显著提高心肌细胞存活率,减少受损细胞心肌酶(LDH、CK)的释放,对心肌细胞具有保护作用。