Computer analysis system of blood oxygen saturation in an animal hypoxia model

An experimental animal hypoxia model has been developed. It consists of two sensors (an in vitro and in vivo model), an experimental device and a computer signal processing system. This method can easily be applied to determine and analyse blood oxygen saturation at various hypoxia levels. It can also be used to evaluate the accuracy of pulse oximetry over a wide range of oxyhemoglobin desaturation levels. The DC and AC components of recorded red and infra-red signals, the dual-wavelength ratio R₁₂ and the reading of a pulse oximeter (SpO₂) can be automatically calculated and displayed on a computer screen. Preliminary results of the animal hypoxia test indicate that the measurements made by the instrument correlate well with the oxygen saturation readings of the automatic blood gas analyser AVL945. The computer analysis system is suitable for repeated estimations in the animal model.     

Endothelin infusion reduces hypoxic pulmonary hypertension in pigs in vivo

Previous work has shown that the plasma levels of the potent vasoactive peptide endothelin (ET) are increased in pathophysiological conditions with increased pulmonary vascular resistance and it has been speculated that ET may play some part in hypoxic pulmonary hypertension. We have therefore evaluated the effects of ET-infusion in the porcine pulmonary circulation after hypoxia-induced hypertension. Pigs under general anaesthesia were artificially ventilated through an endotracheal tube and hypoxia was induced by decreasing the fraction inhaled 0₂ from 0.21 to 0.10. Haemodynamic parameters were continuously recorded using a Swan-Ganz catheter in combination with thermodilution for cardiac output measurements. ET-1 or ET-3 was given as an i.v. infusion through the Swan-Ganz catheter in the right ventricle. Hypoxia induced a reproducible increase in pulmonary vascular resistance (PVR), mean pulmonary artery pressure (MPAP) and right ventricular stroke work (RVSW) while the systemic vascular resistance (SVR) slightly decreased. Cumulative infusion of ET-1 (10, 25 and 50 ng kg^-1 min^-1) dose-dependently decreased MPAP and PVR; at a higher dose (100 ng kg^-1min^-1), the PVR returned to the level observed at hypoxia. ET-infusions at 50 and 100 ng kg^-1 min^-1 evoked an increase in SVR and a decrease in cardiac output (CO) and stroke volume (SV). RVSW also gradually decreased during ET-1 infusion. Infusion of ET-3 evoked effects similar to those of ET-1 infusions, although the response to ET-3 was not that rapid in onset. In a second series of animals, repeated 15 min periods of hypoxia evoked a stable, reproducible response with a consistent increase in PVR, MPAP and RVSW which returned to baseline values during normoxia. Infusion of ET-1 (25 ng kg^-1 min^-1) evoked a rapidly developing decrease in PVR and MPAP which was quickly normalized upon cessation of the ET-infusion. ET-1 infusion at this concentration did not per se influence the haemodynamic parameters during normoxia. It is concluded that in the pig, short-term ET-infusion reduces the pulmonary hypertension associated with acute hypoxia.     

低氧对大鼠肺动脉平滑肌细胞血红素加氧酶-1 mRNA 表达及细胞增殖的影响

探讨血红素加氧酶－1（HO－1）mRNA在低氧大鼠肺动脉平滑肌细胞（PASMC0的表达及HO－1／一氧化碳（HO－1／CO）体系对PASMC增殖的影响。应用荧光定量RT－PCR法测定HO－1mRNA表达。用双波长法检测碳氧血红蛋白（HbCO)吸光值。应用免疫细胞化学方法检测细胞增殖核抗原（PCNA）及核转录因子－κB（NF-κB）的表达。发现HO－1 mRNA在常氧大鼠PASMC有低水平的表达，低氧12h HO-1mRNA水平是常氧时的1．5倍，且HbCO产量随之显著增高（P＜0．01）；低氧24h HO-1 mRNA表达呈回落趋势，HbCO产量亦有所减少，但两者仍高于常氧水平。低氧12h及24h PASMC PCNA核阳性反应颗粒表达较常氧时增强（P＜0．01，P＜0．001），使用HO抑制剂ZnPP-9，其PCNA该阳性反应颗粒表达较单纯低氧时增加更多（P＜0．001，P＜0．01）。低氧组核NF－κB阳性染色较常氧组增强（P＜0．001），使用ZnPP-9，其表达则比低氧时更多（P＜0．01）。低氧通过诱导大鼠PASMC的HO－1 mRNA基因表达，上调HO／CO体系活性，使内源性CO含量增高，抑制PASMC增殖；NF－κB参与了PASMC增殖的调控机制。     

宫内缺氧对新生小鼠额叶皮质神经元内nNOS表达及对成年小鼠学习记忆能力的影响

为探讨孕鼠宫内缺氧对新生小鼠发育时期额叶皮质神经元内nNOS表达及对成年小鼠学习记忆能力的影响,本研究采用Tapanainen建立的缺氧模型致胎龄13、15、17d小鼠宫内缺氧,然后采用Nissl染色观察新生鼠发育时期P1、P7、P14、P28、P90额叶皮质内神经元的数量和形态变化;免疫组织化学染色方法观察新生鼠发育时期P1、P7、P14、P28、P90脑组织内nNOS阳性神经元的表达;Morris水迷宫实验检测P90小鼠的学习和记忆能力。结果显示:与正常组比较,宫内缺氧组小鼠额叶皮质神经元数量明显减少,额叶皮质神经元内nNOS的表达明显减弱。宫内缺氧组小鼠逃避潜伏期延长及穿环次数明显减少。以上结果提示宫内缺氧可导致新生鼠额叶皮质神经元数量明显减少及nNOS的表达也明显减弱,并引起成年小鼠学习记忆能力降低。     

新生鼠缺氧缺血时脑TPA活性与微血管基膜的相关性研究

为了探讨缺氧缺血时脑内组织型纤溶酶原激活物 (TPA)与脑微血管基膜降解的相关性 ,本研究采用了下述二种方法 :第一组是将一日龄 SD大鼠分为五组 :(1)空白对照组 ,(2 )假手术组 ,(3 )缺氧缺血组 ,(4 )缺氧缺血后复氧 2 4h组 ,(5 )缺氧缺血后复氧 48h组 ,每组 12只。每组取 4例测 TPA活性和 8例鼠脑用抗 型胶原、层粘连蛋白和纤维粘连蛋白抗体标记。第二组是脑微血管内皮细胞和星形胶质细胞体外培养 :分为 (1)空白对照组 ,在常规条件下培养的细胞 ;(2 )缺氧组 ,在培养液表面覆盖无菌医用液体石蜡 ,形成缺氧环境 ,每组取 8例培养液测 TPA活性。结果证明 ,在三个实验组中以缺氧缺血组的 TPA活性最高 ,而后随着复氧时间的增加而下降。培养的内皮细胞缺氧组 TPA活性比对照组高 ,而星形胶质细胞缺氧组 TPA活性与对照组无差别。三个实验组的 型胶原、层粘连蛋白和纤维粘连蛋白阳性染色平均单位面积较两对照组者小。三个实验组阳性产物呈不连续线状的微血管数较两对照组多。以上结果显示 ,缺氧缺血可激发新生大鼠脑内 TPA活性增高 ,主要是脑微血管内皮分泌的 TPA活性增高 ,然后通过一系列酶促反应 ,使微血管基膜的细胞外基质成分— 型胶原、层粘连蛋白和纤维粘连蛋白等降解 ,血脑屏障受损 ,微血管的渗透性增?     

低氧增强SY5Y培养细胞内nPKCε的膜转位

目的:通过观察低氧刺激对培养SY5Y神经母细胞瘤细胞内nPKCε和nPKCθ膜转位激活的影响,以探讨两者是否参与细胞低氧预适应的发生。方法:利用本室建立的体外培养SY5Y细胞低氧刺激模型,并应用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白印记(Western-Blot)等方法,半定量分析SY5Y细胞内nPKCε、nPKCθ的膜转位水平。结果:SY5Y细胞经低氧刺激后,随刺激时间(0.5、2、4、6、8、12和24h)的延长,nPKCε在胞浆内的含量逐渐减少,而胞膜成分中的含量则明显增加,且于低氧刺激2h后的增高水平具有统计学显著意义(P<0.001);然而,nPKCθ在正常情况下只存在于胞膜成分内,且其含量不随低氧刺激时间的增加而改变(P>0.1)。结论:nPKCε可能参与了细胞低氧耐受的发生。     

Sympathetic neurons in the superior cervical ganglion are more numerous in SHR and Wistar-Kyoto rats than in wistar rats

Department of Physiology of Man and Animals, Faculty of Biology, Moscow University. Laboratory of Pathophysiology, Research Institue of Pediatrics, Academy of Medical Sciences of the USSR. Department of General Biology, N. I. Pirogov Second Moscow Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR M. Ya. Studenikin.) Translated from Byullet en' Éksperimental'noi Biologii i Meditsiny, Vol. 108, No. 11, pp. 620–622, November, 1989.     

External oxidation pathway in nerve tissue

Institute of Pharmacology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. V. Val'dman.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 107, No. 4, pp. 431–433, April, 1989.     

Upregulation of vascular endothelial growth factor by cobalt chloride-simulated hypoxia is mediated by persistent induction of cyclooxygenase-2 in a metastatic human prostate cancer cell line

Upregulation of vascular endothelial growth factor (VEGF) expression induced by hypoxia is crucial event leading to neovascularization. Cyclooxygenase-2, an inducible enzyme that catalyzes the formation of prostaglandins (PGs) from arachidonic acid, has been demonstrated to be induced by hypoxia and play role in angiogenesis and metastasis. To investigate the potential effect of COX-2 on hypoxia-induced VEGF expression in prostate cancer. We examined the relationship between COX-2 expression and VEGF induction in response to cobalt chloride (CoCl₂)-simulated hypoxia in three human prostate cancer cell lines with differing biological phenotypes. Northern blotting and ELISA revealed that all three tested cell lines constitutively expressed VEGF mRNA, and secreted VEGF protein to different degrees (LNCaP > PC-3 > PC3ML). However, these cell lines differed in the ability to produce VEGF in the presence of CoCl₂-simulated hypoxia. CoCl₂ treatment resulted in 40% and 75% increases in VEGF mRNA, and 50% and 95% in protein secretion by LNCaP and PC-3 cell lines, respectively. In contrast, PC-3ML cell line, a PC-3 subline with highly invasive, metastatic phenotype, exhibits a dramatic upregulation of VEGF, 5.6-fold in mRNA and 6.3-fold in protein secretion after treatment with CoCl₂. The upregulation of VEGF in PC-3ML cells is accompanied by a persistent induction of COX-2 mRNA (6.5-fold) and protein (5-fold). Whereas COX-2 expression is only transiently induced in PC-3 cells and not affected by CoCl₂ in LNCaP cells. Moreover, the increases in VEGF mRNA and protein secretion induced by CoCl₂ in PC-3ML cells were significantly suppressed following exposure to NS398, a selective COX-2 inhibitor. Finally, the effect of COX-2 inhibition on CoCl₂-induced VEGF production was reversed by the treatment with exogenous PGE₂. Our data demonstrate that VEGF induction by cobalt chloride-simulated hypoxia is maintained by a concomitant, persistent induction of COX-2 expression and sustained elevation of PGE₂ synthesis in a human metastatic prostate cancer cell line, and suggest that COX-2 activity, reflected by PGE₂ production, is involved in hypoxia-induced VEGF expression, and thus, modulates prostatic tumor angiogenesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hypoxia aggravates lipopolysaccharide-induced lung injury

The animal model of inflammatory response induced by intratracheal application of lipopolysaccharide includes many typical features of acute lung injury or the acute respiratory distress syndrome. A number of experimental investigations have been performed to characterize the nature of this injury more effectively. In inflammatory conditions, hypoxia occurs frequently before and in parallel with pulmonary and non-pulmonary pathological events. This current study was designed to examine the in vivo effect of hypoxia as a potentially aggravating condition in endotoxin-induced lung injury. Lipopolysaccharide, 150 microg, was instilled intratracheally into rat lungs, and thereafter animals were exposed to either normoxia or hypoxia (10% oxygen). Lungs were collected 2, 4, 6 and 8 h later. Inflammatory response and tissue damage were evaluated by quantitative analysis of inflammatory cells and mediators, surfactant protein and vascular permeability. A significantly enhanced neutrophil recruitment was seen in lipopolysaccharide-animals exposed to hypoxia compared to lipopolysaccharide-animals under normoxia. This increased neutrophil accumulation was triggered by inflammatory mediators such as tumour necrosis factor-alpha and macrophage inflammatory protein-1beta, secreted by alveolar macrophages. Determination of vascular permeability and surfactant protein-B showed enhanced concentrations in lipopolysaccharide-lungs exposed to hypoxia, which was absent in animals previously alveolar macrophage-depleted. This study demonstrates that hypoxia aggravates lipopolysaccharide injury and therefore represents a second hit injury. The additional hypoxia-induced inflammatory reaction seems to be predominantly localized in the respiratory compartment, underlining the compartmentalized nature of the inflammatory response.