The low and high doses administration of lutein improves memory and synaptic plasticity impairment through different mechanisms in a rat model of vascular dementia.

BACKGROUND AND AIM: Vascular dementia (VD) is a common type of dementia. This study aimed to evaluate the effects of low and high doses of lutein administration in bilateral-carotid vessel occlusion (2VO) rats. EXPERIMENTAL PROCEDURE: The rats were divided into the following groups: the control, sham-, vehicle (2VO+V) groups, and two groups after 2VO were treated with lutein 0.5 (2VO+LUT-o.5) and 5mg/kg (2VO+LUT-5). The passive-avoidance and Morris water maze were performed to examine fear and spatial memory. The field-potential recording was used to investigate the properties of basal synaptic transmission (BST), paired-pulse ratio (PPR), as an index for measurement of neurotransmitter release, and long-term potentiation (LTP). The hippocampus was removed to evaluate hippocampal cells, volume, and MDA level. RESULT: Treatment with low and high doses improves spatial memory and LTP impairment in VD rats, but only the high dose restores the fear memory, hippocampal cell loss, and volume and MDA level. Interestingly, low-dose, but not high-dose, increased PPR. However, BST recovered only in the high-dose treated group. CONCLUSIONS: Treatment with a low dose might affect neurotransmitter release probability, but a high dose affects postsynaptic processes. It seems likely that low and high doses improve memory and LTP through different mechanisms.

Gens PSD-95 and GSK-3ß expression improved by hair follicular stem cells-conditioned medium enhances synaptic transmission and cognitive abilities in the rat model of vascular dementia.

INTRODUCTION: Vascular dementia (VaD) is a common type of dementia. The aim of this study was to investigate the cellular and molecular mechanism of conditioned medium (CM) in VaD. MATERIAL AND METHODS: The rats were divided into four groups of control (n = 9), sham-operation (n = 10), VaD with vehicle (n = 9), and VaD with CM (n = 12) that received CM on days 4, 14, and 24 after 2VO. Before sacrificing the rats, cognitive performance was assessed through the open-field (OP), passive-avoidance, and Morris-water maze. The field-potential recording was used to investigate basal synaptic transmission (BST) and long-term potentiation (LTP). Subsequently, the hippocampus was dissected, and real-time PCR was used to quantify the expression levels of ß1-catenin, insulin-like growth factor-1 (IGF-1), transforming growth factor-beta (TGF-ß), glycogen synthase kinase-3ß (GSK-3ß), postsynaptic density protein 95 (PSD-95), and NR2B genes. RESULTS: The results indicated impaired performance in behavioral tests in 2VO rats, coupled with reductions in BST and LTP induction. The expression levels of ß1-catenin, IGF-1, PSD-95, and TGF-ß genes decreased, whereas NR2B and GSK-3ß expression increased. Treatment with CM restores the expression of PSD-95 and GSK-3ß as well as fear-memory, spatial learning, and grooming number without a positive effect on memory retrieval, time spent on the periphery and center of OP. The BST recovered upon administration of CM but, the LTP induction was still impaired. CONCLUSION: The recovery of BST in VaD rats appears to be the most important outcome of this study which is caused by the improvement of gene expression and leads to the restoration of fear memory.

Transmission of high arterial pressure into renal microvessels during venous-clamping augments ischaemia/reperfusion-induced acute kidney injury in anaesthetized rats.

AIM: In two recent studies, we observed that a 30-min renal vein clamping caused formation of interstitial haemorrhagic congestion in ischaemic and ischaemic/reperfused kidney along with the development of severer acute kidney injury (AKI) than renal artery or pedicle clamping. It was suggested that the transmission of high arterial pressure into renal microvessels during vein occlusion probably causes the occurrence of interstitial haemorrhagic congestion that augments AKI. The present investigation aimed to evaluate this suggestion by reducing renal perfusion pressure (RPP) during renal venous occlusion. METHODS: Anaesthetized male Sprague-Dawley rats were divided into three groups (n = 8), which underwent a 2-h reperfusion period following 30-min bilateral renal venous clamping along with reduced RPP (VIR-rRPP group) or without reduced RPP (VIR group) and an equivalent period after sham-operation (Sham group). RESULTS: The VIR-rRPP group compared with VIR group had lower levels of kidney malondialdehyde and tissue damages as epithelial injuries of proximal tubule and thick ascending limb, vascular congestion, intratubular cast and oedema, along with the less reductions in renal blood flow, creatinine clearance, Na+ -reabsorption, K+ and urea excretion, urine osmolality and free-water reabsorption. Importantly, the formation of intensive interstitial haemorrhagic congestion in the VIR group was not observed in the VIR-rRPP group. CONCLUSION: These results indicate that the transmission of high arterial pressure into renal microvessels during venous occlusion leads to rupturing of their walls and the formation of interstitial haemorrhagic congestion, which has an augmenting impact on ischaemia/reperfusion-induced renal structural damages and haemodynamic, excretory and urine-concentrating dysfunctions.

Temporal patterns of alterations in obesity index, lipid profile, renal function and blood pressure during the development of hypertension in male, but not female, rats fed a moderately high-fat diet.

CONTEXT: Male Sprague-Dawley rats consuming a moderately high-fat (MHF)-diet diverge into obesity-prone (OP) with hypertension and obesity-resistant. OBJECTIVES: To study the temporal inter-relationships between body-weight, obesity-index, plasma lipid-profile, renal functional parameters and systolic-pressure alterations during 10-weeks feeding MHF or normal diet to male and female rats. METHODS: Body-weight, obesity-index and systolic-pressure were measured weekly, while metabolic-cage and blood-sampling protocols were performed every other week. After 10-weeks, renal excretory responses to acute salt-loading and renal autoregulation were examined. RESULTS: The male-OP group had progressively increased body-weight, plasma-triglyceride and systolic-pressure from Weeks 2, 4 and 5, respectively, lower renal sodium-excretion at weeks 4-8 and finally, delayed excretory response to salt-loading and rightward and downward shifts in renal autoregulatory curves compared to all other groups. CONCLUSION: Feeding the MHF-diet in male-OP rats led to a greater weight-gain and adiposity followed by the development of atherogenic-hyperlipidaemia and persistently impaired pressure-natriuresis to induce hypertension.

Bilateral renal denervation prevents the development of hypertension during diet-induced obesity in male rats.

NEW FINDINGS: What is the central question of this study? What is the role of the renal nerves in the development of obesity, hyperlipidaemia and hypertension during the long-term feeding of a moderately high-fat diet in male obesity-prone rats? What is the main finding and its importance? The renal nerves play a prominent mediatory role, without influencing the establishment of visceral adiposity and atherogenic hyperlipidaemia, in the induction and progression of pressure natriuresis impairment and hypertension during the developmental period of diet-induced obesity. ABSTRACT: Feeding a moderately high-fat (MHF) diet in male Sprague-Dawley rats induces obesity, pressure natriuresis impairment and hypertension. This study investigated the role of the renal nerves in the impaired pressure natriuresis and hypertension caused by feeding a MHF diet. After collecting baseline data on day 0, 12 rats remained on a low-fat diet (LF group) while the others were switched onto a MHF diet and diverged into obesity-resistant (OR) or obesity-prone (OP). After 4 weeks, half of the OR and OP rats underwent bilateral renal denervation (BRD) to generate four groups: OR, OR/BRD, OP and OP/BRD (n = 12). During 10 weeks, body weight, obesity index, systolic pressure and renal excretory function were measured regularly. After 10 weeks, renal excretory responses to acute salt loading and renal autoregulation were evaluated. The OP and OP/BRD groups had greater increases of body weight and obesity index during the dietary period compared to the other groups, and by week 10 their body weight (425.1 ± 7.2 and 411.9 ± 5.1 g) became considerably larger than that of the LF group (358.5 ± 6.2 g). Renal sodium excretion was reduced by ∼20% at week 4 in the OP and OP/BRD groups, while only the OP group had lower sodium excretion at weeks 6-8 and higher systolic pressure over weeks 5-10 than the other groups and its week 10 systolic pressure reached 138.1 ± 6.7 versus 123.6 ± 2.7 mmHg of the LF group. The OP group showed delayed renal excretory responses to salt loading with rightward and downward shifts in renal autoregulatory curves. Therefore, the renal nerves exert a main mediatory role in the development of pressure natriuresis impairment and hypertension as obesity is established due to the long-term consumption of the MHF diet in male OP rats.

Comparison of ischemic and ischemic/reperfused kidney injury via clamping renal artery, vein, or pedicle in anesthetized rats.

PURPOSE: We recently observed that 30 min of bilateral renal arterial, venous, or pedicle occlusion with 2-h reperfusion differentially induced acute kidney injury (AKI), which was suggested to be probably resulted from their directly exerting dissimilar impacts on kidney during the ischemic period. The present study was further designed to evaluate and prove this suggestion. METHODS: Anesthetized male Sprague-Dawley rats were divided in two distinct supragroups with 30-min bilateral renal ischemia alone (BI) or followed by 30-min reperfusion (BIR), which each had four different groups (n = 8) of subjecting to renal artery, vein, or pedicle clamping and also sham operation. RESULTS: In the BI groups, artery clamping caused lower renal tissue injury than pedicle clamping but vein occlusion caused the highest levels of kidney histological damages along with the widespread hemorrhagic congestion. In the BIR groups, renal vascular congestion, intratubular cast, and edema decreased, but tubular epithelial injury did not significantly change in comparison to their equivalents BI groups. However, the orders of total renal tissue damages in the BIR groups were still as clamping renal veins > > pedicles > arteries and in association with their proportionally developed renal hemodynamic, excretory, and urine-concentrating dysfunctions. CONCLUSION: The transmission of high arterial pressure into renal microvessels and rupturing of their walls during venous-clamping augment, but the retrograde blood flow from veins into kidney during artery clamping attenuates induction of renal tissue injury with respect to pedicle clamping not only at the ischemic period but also at the early reperfusion period and along with the proportional development of renal dysfunctions.

NADPH oxidase 5 activation; a novel approach to human sperm cryoinjury.

Sperm cryopreservation leads to various structural and functional damages, some of which induce by oxidative stress. The reactive oxygen species (ROS) generates by mitochondria and membrane NADPH oxidases (NOXs). Among the NOXs, only NOX5 has been identified in the cell membrane of human sperm. This study was designed to clarify the possible role of NOX5 on sperm cryoinjury. Forty human semen samples were washed and randomly divided into fresh and cryopreserved groups. Each group was divided into 4 subgroups containing Ham's F10 (control), 0.1% DMSO (vehicle), 100 nM of PMA (phorbol 12-myristate 13-acetate) and 1 µM of DPI (diphenyleneiodonium), as NOX5 activator and inhibitor. The samples of cryopreserved groups were preserved in liquid nitrogen for 1 month. The sperm kinematics, membrane integrity, ROS production, apoptosis rate, mitochondrial membrane potential (MMP), intracellular ATP and calcium concentration [Ca2+]i were evaluated. The percent of sperm with intact membrane and motile sperm reduced significantly after thawing (p ≤ 0.01). The ROS production (p ≤ 0.01) and the apoptotic rate increased, MMP dissipated, and the percentage of live cells with high [Ca2+]i decreased significantly in the cryopreserved control group relative to the fresh control group. DPI, in contrast to PMA, improved sperm progressive motility (p ≤ 0.01), membrane integrity in fresh and cryopreserved groups and reduced the ROS amount in cryopreserved group (p ≤ 0.01). Apoptotic rate, [Ca2+]i, ATP, and MMP did not change with DPI and PMA in cryopreserved groups. We conclude that NOX5 activity in fresh sperm is low, and it increases during cryopreservation. NOX5 inhibition improves the cryopreserved sperm quality.

Epidermal neural crest stem cell transplantation as a promising therapeutic strategy for ischemic stroke.

INTRODUCTION: Cell-based therapy is considered as promising strategy to cure stroke. However, employing appropriate type of stem cell to fulfill many therapeutic needs of cerebral ischemia is still challenging. In this regard, the current study was designed to elucidate therapeutic potential of epidermal neural crest stem cells (EPI-NCSCs) compared to bone marrow mesenchymal stem cells (BM-MSCs) in rat model of ischemic stroke. METHODS: Ischemic stroke was induced by middle cerebral artery occlusion (MCAO) for 45 minutes. Immediately after reperfusion, EPI-NCSCs or BM-MSCs were transplanted via intra-arterial or intravenous route. A test for neurological function was performed before ischemia and 1, 3, and 7 days after MCAO. Also, infarct volume ratio and relative expression of 15 selected target genes were evaluated 7 days after transplantation. RESULTS: EPI-NCSCs transplantation (both intra-arterial and intravenous) and BM-MSCs transplantation (only intra-arterial) tended to result in a better functional outcome, compared to the MCAO group; however, this difference was not statistically significant. The infarct volume ratio significantly decreased in NCSC-intra-arterial, NCSC-intravenous and MSC-intra-arterial groups compared to the control. EPI-NCSCs interventions led to higher expression levels of Bdnf, nestin, Sox10, doublecortin, ß-III tubulin, Gfap, and interleukin-6, whereas neurotrophin-3 and interleukin-10 were decreased. On the other hand, BM-MSCs therapy resulted in upregulation of Gdnf, ß-III tubulin, and Gfap and down-regulation of neurotrophin-3, interleukin-1, and interleukin-10. CONCLUSION: These findings highlight the therapeutic effects of EPI-NCSCs transplantation, probably through simultaneous induction of neuronal and glial formation, as well as Bdnf over-expression in a rat model of ischemic stroke.

Renal ischemia/reperfusion induced learning and memory deficit in the rat: Insights into underlying molecular and cellular mechanisms.

Distance organ dysfunction is the major cause of death in the patients with acute kidney injury (AKI). However, the neurobiological basis of AKI-induced brain disorders and their mediators are poorly understood. This study was aimed to find out the links between AKI and brain injury and also the underlying cellular and electrophysiological mechanisms of memory deficit following induction of AKI via different experimental models of renal ischemia with or without uremia and uremia without renal ischemia. Fifty four male Sprague-Dawley rats were divided into 4 groups that underwent 1-h bilateral or 2-h unilateral renal ischemia followed by 1-day reperfusion (BIR and UIR, respectively), and 1-day following bilateral nephrectomy (BNX) or sham-operation. There were 2 subgroups in each group, which blood-brain barrier (BBB) integrity was evaluated in one subgroup. The other subgroup was used for recordings electrophysiological activities of the hippocampus; and after blood sampling and sacrificing animal, the cerebral hemispheres were removed and preserved for performing stereological study and Western-blotting of caspase-3 in the left and right hippocampus, respectively. Plasma urea and creatinine and CA1 neuronal loss were largely increased by BNX and BIR, but slightly by UIR. Apoptosis was stimulated in the hippocampus intensively by BIR but moderately by UIR and BNX. However, BIR and UIR were associated with profoundly disturbed BBB, increased CA1 neuronal excitability, impaired LTP induction and memory deficit. Therefore, AKI most likely through inflammatory mediators leads to hippocampal apoptosis and electrophysiological impairments, BBB disruption and memory loss, whereas uremia may contribute to necrotic neuronal death.

Inhibition of inflammation is not enough for recovery of cognitive impairment in hepatic encephalopathy: Effects of minocycline and ibuprofen.

There is evidence that hyperammonia and inflammation play crucial roles in hepatic encephalopathy. This study intends to determine neuroprotective effects of minocycline (MINO) and ibuprofen (IBU), and also set out to assess whether inhibition of inflammation is enough to achieve optimal improvement of hepatic encephalopathy symptoms. The hepatic encephalopathy was induced by bile-duct ligation (BDL), and the animals received first dose of MINO and/or IBU 15 days later and then every day until the 28 day. The rats were divided into the 6 groups of control, sham, BDL + V and BDL + IBU, BDL + MINO and BDL + MINO + IBU, which each group had 3 sub-groups for evaluations of blood-brain barrier (BBB), memory performance, synaptic-plasticity and apoptosis. The long-term potentiation (LTP) and short-term potentiation were evaluated by field potential recording. The memory performance, apoptosis and BBB integrity were assessed via passive avoidance, Western-blotting of caspase-3 and Evans-blue dye extravasation, respectively. The MINO, IBU or their co-treatment in the BDL rats did not improve liver dysfunction. The BDL increased hippocampal apoptosis and BBB disruption, which were fully recovered by all three pharmacological interventions. The MINO treatment alone or combined with IBU had similar neuroprotective effects on the BDL-induced disturbances of hippocampal basal synaptic transmission, LTP and memory performance, whereas they were not ameliorated by the single IBU therapy. Therefore, it seems likely that inhibition of inflammation is not able to improve functionally impaired memory and LTP in the hepatic encephalopathy, and they may be recovered by the direct neuroprotective effects of the MINO.

Modulation of sphingosine-1-phosphate receptor by FTY720 contributes in improvement of hepatic encephalopathy induced by bile duct ligation.

Hepatic encephalopathy (HE) is a complex neuropsychiatric disorder, which is associated with memory loss and behavioral abnormalities. The cellular and molecular mechanisms that led to hippocampal dysfunction in bile duct ligation (BDL)-induced HE and neuroprotective mechanisms of FTY720 administration were evaluated using whole-cell patch clamp, field-potential recording, western blot, stereology and behavioral experiments. The animals were divided into 4 groups of control (n = 24), sham (n = 21), BDL + V (n = 21) and BDL + FTY720 (n = 20), each having three subgroups. The first subgroup was used for field potential, western blot and stereology experiments. The second and third subgroups were used for behavioral experiments and whole cell patch clamp recording, respectively. The BDL led to considerable loss of hippocampal neurons and apoptosis, along with large impairments in their intrinsic electrophysiological properties, including decrease of firing frequency and increases of first spike latency (FSL), AHP amplitude, irregularity of firing, and half-width, as well as impaired long-term synaptic plasticity and memory. Importantly, FTY720 decreased AHP amplitude probably by direct inhibition of Ca2+ channels and/or KCa2+ currents and improved the FSL and firing irregularity and frequency possibly by decreasing A-type K+ currents in the BDL + FTY720 group. FTY720 administration in the BDL rats also decreased the release probability, which was evident by the increased paired-pulse ratio, but the increased number of readily releasable pool (RRP) of neurotransmitter. Moreover, the AHP improvement and RRP increment most likely led to recovery of LTP and memory performance. In total, FTY720 ameliorated brain disorders in the BDL rats via its direct neuroprotective and/or indirect anti-inflammatory effects.

Effect of CatSper and Hv1 Channel Inhibition on Progesterone Stimulated Human Sperm.

BACKGROUND: Intracellular calcium and proton concentrations are important factors for activating human sperm. Calcium ion (Ca2+) enters sperm through voltage-dependent calcium channel of sperm (CatSper). Proton was extruded from sperm through voltage-gated proton channel (Hv1). In the present study, the selective inhibitors of the CatSper and Hv1 channels, NNC 55-0396 (NNC) and zinc ion, respectively, were used to investigate functions of these channels. METHODS: Normal semen samples (n=24) were washed and diluted to 20×106sperm/ml. The diluted sample was divided into 8 groups, containing Ham's F-10 (the control group), 2 µM NNC, 1 mM ZnCl2 and NNC+Zn. The other 4 groups were the same as above, except that they contained 1 µM progesterone. The computer assisted analysis was done by VT-Sperm 3.1 to determine the percentage of motile sperm and sperm velocity. Acrosomal status was monitored by FITC-PSA and viability assessed by Eosin-Y staining. Statistical comparisons were made using ANOVA followed by Tukey post hoc test. The p<0.05 was considered significant. RESULTS: The percentage of viable and motile sperm, curvilinear velocity and other parameters of motility was reduced in all groups containing NNC, zinc and NNC+ zinc. Progesterone-induced acrosome reaction was abolished by each of these inhibitors. The combination effect of NNC plus zinc on motility and progesterone-induced acrosome reaction was not stronger than NNC by itself. CONCLUSION: CatSper and Hv1 channels play a critical role in human sperm function and viability. It seems that a functional relationship exists between CatSper and Hv1 channels.

Comparison of ischaemia-reperfusion-induced acute kidney injury by clamping renal arteries, veins or pedicles in anaesthetized rats.

NEW FINDINGS: What is central question of this study What are the differences between a traditional renal pedicle-clamping model of acute kidney injury and models with occluded renal artery or vein alone in rats? What is main finding and its importance? During renal venous occlusion, transmission of high arterial pressure into renal capillaries is likely to have caused the rupture of their walls and the occurrence of haemorrhagic congestion that led to higher kidney tissue damage and dysfunction than with pedicle and artery clamping. ABSTRACT: Animal models of ischaemic acute kidney injury (AKI) are valuable tools, but their therapeutic outcomes are not usually translated to humans. Ischaemic AKI in murines is mostly induced via renal pedicle clamping, which is different from patients with AKI that is due to renal artery hypoperfusion or vein thrombosis. This study was designed to compare the traditional pedicle-clamping with artery or vein occlusion alone in rat models of bilateral renal ischaemia-reperfusion (BIR). Twenty-eight anaesthetized male Sprague-Dawley rats were divided into four groups, a sham-operation group and groups that underwent 2 h reperfusion following 30 min clamping of renal arteries (BIR-A group), veins (BIR-V group) or pedicles (BIR-P group). The levels of epithelial injury in proximal tubules and thick ascending limb, intratubular casts and vascular congestion as well as renal malondialdehyde were moderately lower in the BIR-A than BIR-P group, while the BIR-V group showed much higher degrees of these damages than both these groups along with massive haemorrhagic congestion. Accordingly, renal blood flow, glomerular filtration, Na+ reabsorption, K+ and urea excretion, free water reabsorption and urine osmolality were lower in the BIR-V group than in the BIR-A and BIR-P groups, while the BIR-P group had slightly worse renal functional disorders than the BIR-A group. It seems that transmission of high arterial pressure into renal microvessels during venous occlusion causes rupture of capillary walls and haemorrhagic congestion, which leads to intensive kidney injury. In conclusion, the differences in renal disturbances induced by artery, vein and pedicle clamping strongly suggest use of a proper experimental model for each type of human ischaemic AKI.

Pre-Treatment with Erythropoietin Attenuates Bilateral Renal Ischemia-Induced Cognitive Impairments.

One of the most common causes of mortality in acute kidney injury is brain dysfunction. Here we investigated the possible protective effect of erythropoietin (EPO) on cognitive impairments induced by bilateral renal ischemia (BRI). Eighty male Wistar rats were allocated into 8 groups: 1, 2) Sham +V (Vehicle), 3, 4) Sham+EPO, 5, 6) BRI+V, 7, 8) BRI+EPO. The groups followed by the reperfusion periods of 24hours (24 h) and 1week (1w). EPO or saline was administrated 30 min before surgery (1000 IU/kg, i.p). The cognitive function was assessed by passive avoidance learning and Morris water maze tests. Hippocampal brain-derived neurotrophic factor (BDNF) protein expression was assessed by western blotting. BUN (blood urea nitrogen) and creatinine (Cr) concentrations were significantly increased in BRI+V group 24 h after reperfusion. BRI+V rats had just an increased level of BUN but not Cr 1w after reperfusion. EPO reversed passive avoidance learning impairments observed in BRI+V group 24 h after reperfusion. There were no significant differences in spatial and passive avoidance learning between experimental groups 1w after reperfusion and histological evaluation confirmed the behavioral data. BRI significantly decreased the BDNF protein expression in the hippocampus and EPO increased that 24 h after operation. These observations showed protective effect of EPO against cognitive dysfunctions following BRI 24 h after reperfusion through increase in BDNF protein expression.

Long-term treatment of diabetic rats with vanadyl sulfate or insulin attenuate acute focal cerebral ischemia/reperfusion injury via their antiglycemic effect.

It is well-known that patients with diabetes mellitus have worse clinical outcomes following acute ischemic stroke. The intensifying effects of diabetes on ischemic brain injury have been shown to be mostly due to hyperglycemia, rather than the lack of insulin direct effects on brain. It is also well-approved that vanadium compounds have insulin-like and anti-diabetic effects, and the present study was designed to compare the protective effects of diabetes treatment with vanadium or insulin on ischemic/reperfused brain injury. Male Sprague-Dawley rats were divided into 4 groups (n = 21). Two groups of streptozotocin-induced diabetic rats were treated with either vanadyl sulfate or insulin at proper doses to similarly attenuate hyperglycemia during 45 days, while there was no treatment in the control diabetic and non-diabetic sham groups. Thereafter, all treated and non-treated diabetic rats were subjected to 60-min of the right middle cerebral artery occlusion followed by 12-h reperfusion, and then their brains were removed for evaluating blood-brain barrier leakage, tissue swelling, infarct size and oxidant status in both hemispheres. Vanadium and insulin that equally reduced blood glucose and water intake had some differences in their antidiabetic effects of ameliorating weight loss and hypertension during 45-days treatment period. However, they caused similar decrements in levels of Evans blue dye extravastion, edema, infarct volume and malondialdehyde in ischemic/reperfused cerebral hemisphere. Therefore, it can be suggested that insulin and vanadium via their antiglycemic effect cause reduction in cerebral production of oxidants following acute focal ischemia/reperfusion, which attenuate BBB disruption and brain tissue injury.

Cerebral Ischemia-Reperfusion Injuries in Vanadyl-Treated Diabetic Rats.

BACKGROUND: Ischemic stroke recovery is poor in diabetic mellitus (DM). Vanadium compounds (vanadium) relieve DM signs, but their influences on cerebral ischemia/reperfusion injury (I/RI) are inconclusive. Herein, the intensity of I/RI was inspected in vanadium-treated DM rats. METHODS: Rats made diabetic with a single intravenous dose of streptozocin (39 mg/kg). Normal and DM rats used water or vanadyl solution for 45 days. Under isoflurane anesthesia, right middle cerebral artery occlusion was performed for 60 minutes and 12 hours reperfusion. Ischemic rats were divided into untreated-control normal (ICN) and diabetic (ICD), vanadium-treated normal (IVTN) and diabetic (IVTD) groups (n=14 each). After neurological deficit score (NDS) test, the rats were sacrificed and their brain removed and stained with triphenyltetrazolium chloride (TTC) to measure cerebral infarct volume (CIV, mm3) or Evans blue extravasation (EBE, µg/g wet-tissue). Data analysis was performed using one-way ANOVA and Tukey's test (SPSS software, version 21.0) and P values <0.05 were considered statistically significant. RESULTS: Blood glucose (BG, mg/dL) was similar in ICN and IVTN, elevated in IVTD and ICD (245±6 vs. 344±2, P<0.001). The increased CIV in ICN and IVTN was similar (48±2 and 34±5), very high in ICD but lower in IVTD (249±37 vs. 110±16, P<0.001). EBE was absent in non-lesioned hemispheres, similarly increased in lesioned hemispheres of ICN and IVTN (14±1 and 13±1). EBE in IVTD was significantly lower than ICD (21±2 vs. 33±5, P=0.01). CONCLUSION: I/RI was moderate in normoglycemia and did not change with vanadium. Hyperglycemia robustly intensified I/RI. Vanadium ameliorated hyperglycemia and reduced I/RI. Nonetheless, more investigations are required to link the mechanisms of vanadium on DM and stroke injuries.

Structural and functional disorders of hippocampus following ischemia/reperfusion in lower limbs and kidneys.

Recent evidence suggests that ischemia/reperfusion (I/R) in an organ may have distance effect on the brain. In this study, the effects of renal I/R, limb I/R or both together on the structural and function of hippocampus were evaluated and compared. Hence, rats were subjected to 2-h bilateral lower limb ischemia, 45-min bilateral renal ischemia, or combined limb and renal ischemia followed by 1-day reperfusion. At 22-h reperfusion, each rat was fixed on a stereotaxic apparatus for performing electrophysiological study on the hippocampus. The long-term potentiation (LTP) was induced by high-frequency stimulation (HFS), and paired-pulse ratio (PPR) was also monitored before and after HFS delivery. After taking blood sample and sacrificing animal, its brain was removed and preserved for stereological study. The limb I/R reduced plasma osmolality that led to synaptic excitement in the hippocampus, where there was a considerable loss of pyramidal cells and markedly impaired short- and long-term synaptic plasticity. The renal I/R largely increased plasma creatinine that might excite basal synaptic transmission. In the rats with combined limb and renal I/R, the hippocampal neuronal loss and impaired synaptic plasticity were the same as those with limb I/R, but basal synaptic transmission was lowered. In conclusion, the 2-h lower limb ischemia compared to 45-min renal ischemia induced more injurious distant effects on the hippocampus after 1-day reperfusion. The combination of renal and limb I/R did not add or potentiate hippocampal neuronal loss and synaptic plasticity impairment, whereas it decreased the basal synaptic transmission with respect to each one alone.

The Interaction between Trolox and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic Acid on Hypoxic Pulmonary Vasoconstriction in the Isolated Rabbit Lung.

BACKGROUND: The mechanism of hypoxic pulmonary vasoconstriction (HPV) is still debatable. It has been proposed that reactive oxygen species (ROS) might be involved in HPV. However, there is no special transporter for superoxide anion in the cell membrane and it may release from the cells via anion exchanger. Therefore, the aim of this study was to investigate the interaction of ROS and anion exchanger in acute HPV. METHODS: The present study was performed in the isolated rabbit lung. After preparation, the lungs were divided into four hypoxic groups of control, Trolox (antioxidant)-treated, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS, anion exchanger inhibitor)-treated, and Trolox+DIDS-treated. Pulmonary artery pressure, left atrial pressure, and lung weight were continuously registered and PVR was then calculated. PO2, PCO2, HCO3-, pH, and NO metabolites of the perfusate were measured during steady-state and at the end of experiments (30 minutes). All data were compared with ANOVA and t-test and significance was considered when P<0.05. RESULTS: Ventilation of the lungs with hypoxic gas induced HPV in the control group. DIDS did not have a further effect on HPV compared with the control group. The combination of Trolox and DIDS decreased HPV rather than Trolox per se at 5 minutes. Furthermore, HPV was abolished in both the Trolox and Trolox+DIDS groups at 30 minutes. Concentrations of NO metabolites in the Trolox+DIDS group were more than other groups. CONCLUSION: The present study indicates a possible interaction between ROS and anion exchanger in acute HPV. It also suggests the modulatory effect of NO at above condition.

Netrin-1 improves the amyloid-ß-mediated suppression of memory and synaptic plasticity.

The aim of this study was to investigate the effects of netrin-1 (NT-1) on amyloid-beta (Aß)-induced impairments in learning-memory and synaptic plasticity. The NT-1 or its vehicle was administered four times into the Aß+NT or Aß+V groups, respectively. The Aß+SNT group received a single dose of NT-1. The Aß+HNT group received heat-inactivated NT-1. For the learning-memory and synaptic plasticity assessment, field potentials recording and behavioral experiment were used. Bilateral injection of Aß1-42 inhibits induction of long-term potentiation (LTP) and decreased memory performance in all the behavioral tasks. However, only by repeated injection of NT-1, significant recovery of LTP and memory was seen. Although, the delivery of HFS to Aß+NT group recovered EPSP slope of the maintenance phase when compared with Aß+V, but it failed to recover the induction phase. It can be assumed that NT-1 may have regulatory effects on the synthesis of key proteins and/or structural changes that are responsible for LTP induction, since the protein synthesis and/or structural changes are required for the maintenance phase of LTP.

Renal ischemia/reperfusion against nephrectomy for induction of acute lung injury in rats.

PURPOSE: Acute kidney injury (AKI) induces acute lung injury (ALI) through releasing injurious mediators or impairing clearance of systemic factors. To determine the links between AKI and ALI, pulmonary and blood variables were evaluated following induction of AKI via different experimental models of bilateral renal ischemia/reperfusion (BIR: renal ischemia with uremia), unilateral renal ischemia/reperfusion (UIR: renal ischemia without uremia), bilateral nephrectomy (BNX: uremia without renal ischemia), and unilateral nephrectomy (UNX: without uremia and renal ischemia). METHODS: Ninety male Sprague-Dawley rats were divided into six groups. Animals had 1-h bilateral or 2-h unilateral renal ischemia followed by 24-h reperfusion in the BIR and UIR groups, respectively, and 24-h period following bilateral or unilateral nephrectomy in the BNX and UNX groups, respectively. There were also sham and control groups with and without sham-operation, respectively. RESULTS: Plasma malondialdehyde and nitric oxide were elevated by BIR more than UIR, but not changed by UNX and BNX. UIR slightly increased plasma creatinine, whereas BIR and BNX largely increased plasma creatinine, urea, K+ and osmolality and decreased arterial HCO3-, pH, and CO2. UNX and UIR did not affect lung, but BIR and BNX induced ALI with equal capillary leak and macrophages infiltration. However, there were more prominent lung edema and vascular congestion following BNX and more severe neutrophils infiltration and PaO2/FiO2 reduction following BIR. CONCLUSION: Acutely accumulated systemic mediators following renal failure in the absence of kidneys vary from those due to combined renal failure with ischemic-reperfused kidneys and consequently they induce ALI with distinct characteristics.