Disruption of a single copy of the p38alpha MAP kinase gene leads to cardioprotection against ischemia-reperfusion

The p38 mitogen-activated protein kinase (p38) is activated in the heart during ischemia-reperfusion. However, it is not clear whether the activation of p38 is the protective response or the kinase mediates the cellular damage by ischemia-reperfusion. We examined the role of p38alpha in ischemia-reperfusion injury by studying p38alpha(+/-) mice. The p38alpha protein level in the p38alpha(+/-) heart was 50+/-8.7% compared with that in the p38alpha(+/+) heart. Upon reperfusion following ischemia for 25min, p38alpha activity was transiently increased. The maximum level of p38 activity in p38alpha(+/-) was 60+/-10.5% compared with that in p38alpha(+/+). In the p38alpha(+/+) heart, 25min ischemia and 2h reperfusion resulted in necrotic injury (37.1+/-2.7% of the area at risk), whereas infarct size was drastically reduced to 7.2+/-0.7% in the p38alpha(+/-) heart. These suggested that p38alpha plays a pivotal role in the signal transduction pathway mediating myocardial cell death caused by ischemia-reperfusion.     

Protective activity of (-)-epicatechin 3-O-gallate against peroxynitrite-mediated renal damage

The protective effect of ( -)-epicatechin 3- O -gallate (ECg) against peroxynitrite (ONOO -)-mediated damage was examined using an animal model and a cell culture system. In rats subjected to lipopolysaccharide (LPS) administration plus ischemia-reperfusion, the plasma 3-nitrotyrosine level, an indicator of ONOO - production in vivo , was elevated, whereas it declined significantly and dose-dependently after the oral administration of ECg at doses of 10 and 20 μmoles/kg body weight/day for 20 days prior to the process. Moreover, oral administration of ECg significantly enhanced the activities of the antioxidant enzymes, superoxide dismutase, catalase and glutathione peroxidase, and the antioxidant glutathione, showing enhancement of the biological defense system against the damage induced by ONOO -. In addition, the significant increase in the renal mitochondrial thiobarbituric acid-reactive substance level of LPS and ischemic-reperfused control rats was attenuated in rats given ECg. Furthermore, the elevations in the plasma urea nitrogen and creatinine (Cr) levels and the urinary methylguanidine/Cr ratio induced by the procedure were attenuated markedly after oral administration of ECg, implying amelioration of renal impairment. The addition of ECg (25 or 125 μM) prior to 3-morpholinosydnonimine (SIN-1, 800 μM) exposure reduced ONOO - formation and increased the viability of cultured renal epithelial (LLC-PK 1 ) cells in a dose-dependent manner. In particular, ECg inhibited ONOO --mediated apoptotic cell death, which was confirmed by decreases in the DNA fragmentation rate and the presence of apoptotic morphological changes, i.e. small nuclei and nuclear fragmentation. Furthermore, adding ECg before SIN-1 treatment regulated the cell cycle by enhancing G 2 /M phase arrest. This study provides evidence that ECg has protective activity against the renal damage induced by excessive ONOO - in cellular and in vivo systems.     

电子顺磁共振法测定清栓酶在沙鼠脑缺血再灌流损伤中的抗自由基作用

本文设计了沙鼠脑缺血再灌流实验模型,以清拴酶作脑缺血后静脉注射再灌流模型和非药物再灌流模型组对照,用电子顺磁共振法(ESR)测定脑组织自由基波谱,结果清栓酶组出现自由基信号很弱,接近正常组,显著区别于未用药脑缺血再灌流30分钟和60分钟组(P<0.01),证明清栓酶有抗自由基和再灌注损伤的作用.     

Detection of ischemia-reperfusion cardiac injury by cardiac muscle chemiluminescence

Various methods have been used in the past to assess the implication of oxygen free radicals (OFR) in ischemia-reperfusion-induced cardiac injury. Luminol-enhanced tert-butyl-initiated chemiluminescence in cardiac tissue reflects oxidative stress and is a very sensitive method. It was used to elucidate the role of OFR in cardiac injury due to ischemia and reperfusion. Studies were conducted on perfused isolated rabbit hearts in three groups (n = 8 in each): I, control; II, submitted to global ischemia for 30 min; III, submitted to ischemia for 30 min followed by reperfusion for 60 min. The heart tissue was then assayed for chemiluminescence (CL); content of malondialdehyde (MDA), an indicator of OFR-induced cardiac injury; and activity of tissue levels of antioxidants [superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px)].The control values for left and right ventricular CL and malondialdehyde were 81.1 ± 15.4 (S.E.) and 182.4 ± 50.3 (S.E.), mv-min-mg protein^–1; and 0.024 ± 0.006 (S.E.) and 0.324 ± 0.005 (S.E.) nmoles-mg protein^–1 respectively. Ischemia produced an increase in the cardiac CL (3.3 to 4.4 fold) and MDA content (2 to 2.6 fold). Reperfusion following ischemia also produced similar changes in CL and MDA content. The control values for activity of left ventricular SOD, catalase, and GSH-Px were 45.77 ± 1.73 (S.E.) U-mg protein^–1 5.35 ± 0.51 (S.E.) K-10^–3-sec^–1-mg protein^–1, and 77.50 ± 7.70 (S.E.) nmoles NADPH-min^–1-mg protein^–1 respectively. Activities of SOD and catalase decreased during ischemia but were similar to control values in ischemic-reperfused hearts. The GSH-Px activity of left ventricle was unaffected by ischemia, and ischemia-reperfusion. GSH-Px activity of the right ventricle increased with ischemia, and ischemic-reperfusion.These results indicate that cardiac tissue chemiluminescence would be a useful and sensitive tool for the detection of oxygen free radical-induced cardiac injury.     

Overexpression of Cu,Zn superoxide dismutase attenuates oxidative inhibition of astrocyte glutamate uptake

Glutamate neurotoxicity in brain is normally prevented by rapid uptake of glutamate by astrocytes. Increased expression of Cu,Zn superoxide dismutase (SOD1) can increase resistance to cerebral ischemia and other oxidative insults, but the cellular mechanisms by which this occurs are not well established. Here we examine whether increased SOD1 expression can attenuate inhibition of astrocyte glutamate uptake by reactive oxygen species. Primary cortical astrocyte cultures were prepared from transgenic mice that overexpress human SOD1 and from nontransgenic littermate controls. Glutamate uptake was assessed after exposure of these cultures to xanthine oxidase plus hypoxanthine, an extracellular superoxide generating system, or to menadione, which generates superoxide in the cytosol. These treatments produced dose-dependent reductions in astrocyte glutamate uptake, and the reductions were significantly attenuated in the SOD1 transgenic astrocytes. A specific effect of reactive oxygen species on glutamate transporters was suggested by the much smaller inhibitory effects of xanthine oxidase/hypoxanthine and menadione on GABA uptake than on glutamate uptake. These findings suggest that the cerebroprotective effects of increased SOD1 expression during cerebral ischemia-reperfusion could be mediated in part by astrocyte glutamate transport.     

Effect of ellagic acid on gastric damage induced in ischemic rat stomachs following ammonia or reperfusion

We examined the effect of ellagic acid (EA), one of the polyphenols that are abundantly contained in whisky as a nonalcoholic component, on gastric lesions induced by ammonia plus ischemia or ischemia/reperfusion in rats, in relation to the antioxidative system. Under urethane anesthesia, a rat stomach was mounted in an ex vivo chamber, and the following two experiments were performed; 1) a stomach was made ischemic (1.5 ml/100 g body weight) for 20 min, followed by reperfusion for 15 min in the presence of 100 mM HCl; 2) a stomach was made ischemic by bleeding from the carotid artery (1 ml/100 g body weight), followed by intragastric application of ammonia (NH4OH: 120 mM). EA (0.1-10 mg/ml) was applied in the chamber 30 min before the onset of ischemia. Gastric potential difference (PD) and mucosal blood flow (GMBF) were measured before, during and after 20 min of ischemia. Ischemia/reperfusion caused a profound drop in GMBF followed by a return, and resulted in hemorrhagic lesions in the stomach in the presence of 100 mM HCI. These lesions were dose-dependently prevented by EA with suppression of lipid peroxidation but no effect on GMBF, and the effect at 6 mg/ml was almost equivalent to that of superoxide dismutase (SOD: 15000 unit/kg/hr) infused i.v. during a test-period. On the other hand, application of NH4OH to the ischemic stomach produced a marked reduction in PD, resulting in severe hemorrhagic lesions. These changes were prevented with both EA and SOD. In addition, EA had a potent scavenging action against monochloramine in vitro. These results suggest that EA exhibits gastric protective action against gastric lesions induced by NH4OH or reperfusion in the ischemic stomach, probably due to its anti-oxidative activity. This property of EA partly explains the less damaging effect of whisky in the stomach and may be useful as the prophylactic for Helicobacter pylori-associated gastritis.     

胸段硬膜外阻滞对家猪肝缺血再灌注时微循环的影响

目的：探讨胸段硬膜外阻滞对家猪肝缺血再灌注时微循环的影响。方法：14只健康幼年家猪,随机分为两组：硬膜外组（E组）和对照组（C组）,每组7只。基础麻醉后,进行气管插管机械通气,侧卧位行胸段硬膜外穿刺（T8-9）置管。经腹正中切口暴露肝脏,分离肝十二肠韧带,游离肝左动脉及门静脉左支备用。E组给予0．5％布比卡因0．75mL／节段（阻滞T5-T12脊神经）,C组给予相同容积的生理盐水。硬膜外给药15min后,夹闭肝左动脉及门静脉左支45min,然后开放肝左动脉及门静脉左支进行再灌注4h。分别在缺血前（TO）,缺血末（T1）,再灌注1h（T2）,2h（T3）,4h（T4）采血样用于测定AST,ALT,NO,ET-1,TNF-α,IL-1β和ATP。实验结束后取肝脏组织样本测定MDA和观察肝脏病理变化。结果：E组各时点MAP水平低于C组,有统计学差异（P〈0．05）;pH值以及PaC02两组间无明显差异;E组各时点AST,ALT,ET-1,TNF-α,IL-1β浓度低于C组,有统计学差异（P〈0．05）,NO及ATP浓度高于C组,有统计学差异（P〈0．05）;MDA含量c组高于E组,有统计学差异（P〈O．05）。光镜下可见E组肝组织损伤程度轻于C组。结论：胸段硬膜外阻滞可以增加肝脏能量的生成,纠正NO／ET的失衡,减少促炎介质的生成,从而改善微循环障碍的程度减轻肝缺血再灌注损伤。     

Prevention of Peroxynitrite-induced Renal Injury through Modulation of Peroxynitrite Production by the Chinese Prescription Wen-Pi-Tang

The effect of Wen-Pi-Tang extract on renal injury induced by peroxynitrite (ONOO &#109 ) production was investigated using rats subjected to intravenous lipopolysaccharide (LPS) injection and then renal ischemia followed by reperfusion. The plasma level of 3-nitrotyrosine, a marker of cytotoxic ONOO &#109 formation in vivo, was enhanced markedly in control rats subjected to LPS plus ischemia-reperfusion, but was significantly reduced by the oral administration of Wen-Pi-Tang extract, at doses of 62.5 and 125 mg/kg body weight/day, for 30 days prior to LPS plus ischemia-reperfusion. The activities of inducible nitric oxide synthase (iNOS) and xanthine oxidase (XOD) in renal tissue of control and Wen-Pi-Tang extract-treated rats did not change significantly, while those of the antioxidant enzymes, superoxide dismutase, catalase and gluta-thione peroxidase, were significantly increased by the administration of Wen-Pi-Tang extract, indicating that Wen-Pi-Tang improved the defense system by scavenging free radicals, not by directly inhibiting nitric oxide and superoxide production by iNOS and XOD. In addition, the levels of the hydroxylated products, m - and p -tyrosine, declined, whereas that of phenylalanine increased, after oral administration of Wen-Pi-Tang extract. Furthermore, the elevated plasma urea nitrogen and creatinine levels resulting from LPS plus ischemia-reperfusion process were significantly reduced by Wen-Pi-Tang extract, implying amelioration of renal impairment. The present study indicates that Wen-Pi-Tang extract contributes to the regulation of ONOO &#109 formation and plays a beneficial role against ONOO &#109 -induced oxidative injury and renal dysfunction in vivo .     

Adenosine deaminase in ischemia reperfusion injury in patients with myocardial infarction

A comparative study on the levels of erythrocyte adenosine deaminase and lipid peroxidation has been undertaken in patients with myocardial infarction before and after thrombolysis along with matched healthy individuals. Our findings show that adenosine deaminase activity is highly elevated in post-reperfused patients when compared to pre- thrombolysed and healthy persons. Malondialdehyde(MDA) levels are also significantly increased in post-thrombolysed patients. The study reveals an important role of adenosine deaminase in reperfusion injury in patients with myocardial infarction.     

人参皂甙Rb1对大鼠脑缺血再灌注神经损伤后的修复作用

通过阻塞大鼠大脑动脉,制备短暂性脑缺血大鼠模型,将出现神经功能缺失症状的大鼠随机分组,实施再灌注后立即按(40mg/kg)进行腹腔注射人参皂甙Rb1。结果发现大鼠脑缺血再灌注后,人参皂甙Rb1通过促进NAIP、Bcl-2表达和抑制Bax表达发挥神经损伤后的修复作用。人参皂甙Rb1给药组在各时间点的胶质细胞源性神经营养因子(GDNF)阳性细胞数远远高于单纯脑缺血再灌注组(P<0.01)。GDNF的表达与缺血性损伤有一定的联系,可认为人参皂甙Rbl对神经系统有一定保护作用。