Efficacy of Acetylcysteine and Selenium in Aneurysmal Subarachnoid Hemorrhage Patients: A Prospective, Multicenter, Single Blind Randomized Controlled Trial.

BACKGROUND: Subarachnoid hemorrhage (SAH) patients have oxidative stress results in inflammation, tissue degeneration and neuronal damage. These deleterious effects cause aggravation of the perihematomal edema (PHE), vasospasm, and even hydrocephalus. We hypothesized that antioxidants may have a neuroprotective role in acute aneurysmal SAH (aSAH) patients. METHODS: We conducted a prospective, multicenter randomized (single blind) trial between January 2017 and October 2019, investigating whether antioxidants (acetylcysteine and selenium) have the potential to improve the neurologic outcome in aSAH patients. The antioxidant patient group received antioxidants of acetylcysteine (2,000 mg/day) and selenium (1,600 µg/day) intravenously (IV) for 14 days. These drugs were administrated within 24 hours of admission. The non-antioxidant patient group received a placebo IV. RESULTS: In total, 293 patients were enrolled with 103 patients remaining after applying the inclusion and exclusion criteria. No significant differences were observed in the baseline characteristics between the antioxidant (n = 53) and non-antioxidant (n = 50) groups. Among clinical factors, the duration of intensive care unit (ICU) stay was significantly shortened in patients who received antioxidants (11.2, 95% confidence interval [CI], 9.7-14.5 vs. 8.3, 95% CI, 6.2-10.2 days, P = 0.008). However, no beneficial effects were observed on radiological outcomes. CONCLUSION: In conclusion, antioxidant treatment failed to show the reduction of PHE volume, mid-line shifting, vasospasm and hydrocephalus in acute SAH patients. A significant reduction in ICU stay was observed but need more optimal dosing schedule and precise outcome targets are required to clarify the clinical impacts of antioxidants in these patients. TRIAL REGISTRATION: Clinical Research Information Service Identifier: KCT0004628.

Reactive Oxygen Species Scavenger in Acute Intracerebral Hemorrhage Patients: A Multicenter, Randomized Controlled Trial.

[Figure: see text].

Lower Serum n-3 Fatty Acid Level in Older Adults with Sarcopenia.

The n-3 fatty acid (FA) has evoked considerable interest as a modifiable factor for maintenance of muscle health owing to its anti-inflammatory properties. To clarify this possibility, we investigated circulating n-3 FA level, a reliable biomarker of FA status in the body, in relation to sarcopenia in a cohort of Asian older adults. Blood samples were collected from 125 participants who underwent comprehensive assessment of muscle mass and function. Serum FA level was measured by gas chromatography/mass spectrometry. Sarcopenia was diagnosed using the cut-off points specified for the Asian population. After adjusting for sex, age, and body mass index, subjects with sarcopenia and those with low muscle strength had 36.5% and 32.4% lower serum n-3 levels (P = 0.040 and 0.030), respectively, than controls. The odds ratios per standard deviation increment in serum n-3 level for sarcopenia and low muscle strength were 0.29 and 0.40 (P = 0.015 and 0.028), respectively. A higher serum n-3 level was significantly associated with greater muscle strength (P = 0.038). These findings suggest a possible protective effect of n-3 FA on human muscle homeostasis. Further well-designed large-scale longitudinal studies are necessary to understand the definite role of circulating n-3 FA level in sarcopenia risk assessment.

Dll4 Suppresses Transcytosis for Arterial Blood-Retinal Barrier Homeostasis.

RATIONALE: Central nervous system has low vascular permeability by organizing tight junction (TJ) and limiting endothelial transcytosis. While TJ has long been considered to be responsible for vascular barrier in central nervous system, suppressed transcytosis in endothelial cells is now emerging as a complementary mechanism. Whether transcytosis regulation is independent of TJ and its dysregulation dominantly causes diseases associated with edema remain elusive. Dll4 signaling is important for various vascular contexts, but its role in the maintenance of vascular barrier in central nervous system remains unknown. OBJECTIVE: To find a TJ-independent regulatory mechanism selective for transcytosis and identify its dysregulation as a cause of pathological leakage. METHODS AND RESULTS: We studied transcytosis in the adult mouse retina with low vascular permeability and employed a hypertension-induced retinal edema model for its pathological implication. Both antibody-based and genetic inactivation of Dll4 or Notch1 induce hyperpermeability by increasing transcytosis without junctional destabilization in arterial endothelial cells, leading to nonhemorrhagic leakage predominantly in the superficial retinal layer. Endothelial Sox17 deletion represses Dll4 in retinal arteries, phenocopying Dll4 blocking-driven vascular leakage. Ang II (angiotensin II)-induced hypertension represses arterial Sox17 and Dll4, followed by transcytosis-driven retinal edema, which is rescued by a gain of Notch activity. Transcriptomic profiling of retinal endothelial cells suggests that Dll4 blocking activates SREBP1 (sterol regulatory element-binding protein 1)-mediated lipogenic transcription and enriches gene sets favorable for caveolae formation. Profiling also predicts the activation of VEGF (vascular endothelial growth factor) signaling by Dll4 blockade. Inhibition of SREBP1 or VEGF-VEGFR2 (VEGF receptor 2) signaling attenuates both Dll4 blockade-driven and hypertension-induced retinal leakage. CONCLUSIONS: In the retina, Sox17-Dll4-SREBP1 signaling axis controls transcytosis independently of TJ in superficial arteries among heterogeneous regulations for the whole vessels. Uncontrolled transcytosis via dysregulated Dll4 underlies pathological leakage in hypertensive retina and could be a therapeutic target for treating hypertension-associated retinal edema.

The effects of daidzin and its aglycon, daidzein, on the scopolamine-induced memory impairment in male mice.

In this study, the effect of daidzin or daidzein isolated from Pueraria lobata on the memory impairments induced by scopolamine was assessed in male mice using the passive avoidance and the Morris water maze tasks. Administration of daidzin (5 mg/kg) or daidzein (5 mg/kg) significantly reversed the scopolamine (1 mg/kg)-induced cognitive impairments in male mice as evidenced by the passive avoidance test (p < 0.05) and on the Morris water maze test (p < 0.05). Moreover, the ameliorating effects of daidzin or daidzein were antagonized by tamoxifen (1 mg/kg), the nonspecific estrogen receptor antagonist. These results indicate that daidzin or daidzein may be useful in cognitive impairment induced by cholinergic dysfunction, and this beneficial effect is mediated, in part, via estrogen receptor.

Anti-amnesic effect of ESP-102 on Aß(1-42)-induced memory impairment in mice.

The aim of this study was to characterize the effects of ESP-102 on the memory impairments and pathological changes induced by amyloid-ß (Aß)(1-42) peptide in mice. The ameliorating effect of ESP-102 on memory impairment was investigated using the passive avoidance and the Morris water maze tasks, and the pathological changes were identified by immunohistochemistry and western blotting. Aß(1-42) peptide (3µg/3µl) was administered by intracerebroventricular injection. By the single administration of ESP-102 (100mg/kg, p.o), the memory impairment induced by Aß(1-42) peptide was significantly attenuated (P<0.05). Moreover, ESP-102 (100mg/kg, p.o) significantly inhibited acetylcholinesterase (AChE) activity in the hippocampus compared to the Aß(1-42) peptide-injected control group. In the subchronic treatment study, ESP-102 (50 or 100mg/kg/day, p.o) administration for seven days ameliorated the memory impairments induced by Aß(1-42) peptide. Moreover, ESP-102 inhibited lipid peroxidation induced by Aß(1-42) peptide in the hippocampus. Aß(1-42)-induced increases in the expression of GFAP (an astrocyte marker) and inducible nitric oxide synthase (iNOS) in the hippocampal region were also attenuated by ESP-102 treatment. These results suggest that the ameliorating effect of ESP-102 on Aß(1-42) peptide-induced memory impairment is mediated via its AChE inhibitory, antioxidative, and/or anti-inflammatory activities.

Antidepressant-like activity of the aqueous extract of Allium macrostemon in mice.

AIM OF THE STUDY: The aim of this study was to identify the effects of water extracts of Allium macrostemon Bunge (AM-W), a traditional herb, in mice. MATERIALS AND METHODS: The antidepressant-like activities of AM-W were evaluated through behavioral despair in forced swimming test and tail suspension test. To elucidate the mode of action of the antidepressant-like effects of AM-W, new born cells in the subgranular zone and the granule cell layer were analyzed by immunostaining for incorporation of 5-bromo-2-deoxyuridine (BrdU). In addition, the effects of AM-W on the expression levels of brain-derived neurotrophic factor (BDNF) were investigated by western blotting and immunohistochemistry. RESULTS: The administration of AM-W reduced the immobility duration in the forced swimming test and tail suspension test (100 or 200 mg/kg, P<0.05). Sub-chronic administration of AM-W (100 or 200 mg/kg, p.o., for 14 days) increased the number of BrdU-incorporating cells. The percentage of BrdU-incorporating cells co-localized with NeuN was significantly increased after AM-W administration (100 or 200 mg/kg, P<0.05). Moreover, the expression levels of BDNF which is reported to be associated with neurogenesis were significantly increased in the hippocampus after administration of AM-W. CONCLUSIONS: These results suggest that AM-W may be a good antidepressant, and that its mechanism of action may be related to its positive effects on neurogenesis and BDNF release.

The memory-enhancing effects of Euphoria longan fruit extract in mice.

AIM OF THE STUDY: The fruit of Euphoria longan (Lour.) Steud. (Sapindaceae) is sweet and edible. Dried Euphoria longan fruit is prescribed as a tonic and for the treatment of forgetfulness, insomnia, or palpitations caused by fright in traditional Chinese medicine. The effects of aqueous extract of Euphoria longan fruit (ELE) on learning and memory and their underlying mechanisms were investigated. MATERIALS AND METHODS: Aqueous extract of Euphoria longan fruit (ELE) was administered to ICR mice for 14 days. Piracetam was used as a positive control for its known memory-enhancing effects. Memory performances were assessed using the passive avoidance task. The expressions of phosphorylated extracellular signal-regulated kinase (pERK) 1/2, phosphorylated cAMP response element binding protein (pCREB), brain-derived neurotrophic factor (BDNF), doublecortin (DCX) and the incorporation of 5-bromo-2-deoxyuridine (BrdU) in hippocampal dentate gyrus and CA1 regions were investigated using immunohistochemical methods. RESULTS: The step-through latency in the ELE-treated group was significantly increased compared with that in the vehicle-treated controls (P<0.05) in the passive avoidance task. Piracetam-treated group also showed enhanced cognitive performaces in the passive avoidance task. Immunohistochemical studies revealed that the number of cells immunopositive for BDNF, pCREB, or pERK 1/2 was significantly increased in the hippocampal dentate gyrus and CA1 regions after ELE treatment for 14 days (P<0.05). DCX and BrdU immunostaining also revealed that ELE significantly enhanced immature neuronal survival, but not neuronal cell proliferation in the subgranular zone of the dentate gyrus. CONCLUSIONS: The present results suggest that subchronic administration of aqueous extract of Euphoria longan fruit enhances learning and memory, and that its beneficial effects are mediated, in part, by BDNF expression and immature neuronal survival.

Tanshinone I enhances learning and memory, and ameliorates memory impairment in mice via the extracellular signal-regulated kinase signalling pathway.

BACKGROUND AND PURPOSE: The intracellular signalling kinase, extracellular signal-regulated kinase 1/2 (ERK1/2) is required for new memory formation, suggesting that control of ERK signalling might be a target for the treatment of cognitive dysfunction. Previously, we reported that tanshinone congeners have ameliorating effects on drug-induced memory impairment in mice. Here, we have investigated possible modes of action of tanshinone I on learning and memory, associated with ERK phosphorylation. EXPERIMENTAL APPROACH: Using immunohistochemical, Western blot techniques, and behavioural testing, we studied the effect of tanshinone I on memory impairment induced by diazepam or dizocilpine (MK-801) in mice. KEY RESULTS: Tanshinone I (2 or 4 mg.kg(-1), p.o.) increased latency times versus vehicle-treated control group in the passive avoidance task. Western blot analysis and immunohistochemical data showed that tanshinone I (4 mg.kg(-1)) increased levels of phosphorylated cAMP response element binding protein (pCREB) and phosphorylated ERK (pERK) in the hippocampus. These increases in pCREB and pERK were blocked by U0126 (inhibitor of ERK1/2), which also prevented the increase in passive avoidance task latency time after tanshinone I. In models of learning and memory impairment induced by diazepam and MK-801, tanshinone I (4 mg.kg(-1)) reversed learning and memory impairments detected by the passive avoidance test. Western blot analysis showed that tanshinone I reversed the diazepam- and MK-801-induced inhibitions of ERK and CREB activation in hippocampal tissues. These effects were also blocked by U0126. CONCLUSIONS AND IMPLICATIONS: Tanshinone I ameliorates the learning and memory impairments induced by diazepam and MK-801 through activation of ERK signalling.

Chronic hypoperfusion increases claudin-3 immunoreactivity in rat brain.

Chronic hypoperfusion-induced changes in blood-brain barrier (BBB) tight junction components have not been well studied. In the present study, we investigated the temporal profiles of claudin-3 (a BBB tight junction element) and myleoperoxidase (MPO, a marker of neutrophil infiltration) in the cortical and thalamic regions of rat brain subjected to chronic cerebral hypoperfusion. Chronic cerebral hypoperfusion was induced by an occlusion of two common carotid arteries and the immunoreactivity of claudin-3 or MPO was determined at 1, 2, 3, or 6 weeks after the occlusion. A typical pattern of BBB breakdown was observed from 2 weeks of the occlusion in cortical and thalamic regions based on Evans Blue leakage. Claudin-3 immunoreactivity was increased only in cortical regions after 2 weeks of occlusion. However, after 3 weeks of occlusion, marked increases in claudin-3 immunoreactivity were observed in both cortical and thalamic regions (P<0.05), which persisted for at least 6 weeks after the occlusion despite a slight reduction. In contrast, MPO immunoreactivity was increased only in the thalamic regions after 2 weeks of occlusion. But the pattern of MPO immunoreactivity at 3 and 6 weeks after the occlusion was same as claudin-3. At these time points, MPO immunoreactivity was significantly increased in both cortical and thalamic regions (P<0.05). These results show that chronic cerebral hypoperfusion increases the immunoreactivity of claudin-3 and neutrophil infiltration in cortical and thalamic regions of the brain, and demonstrate changes in BBB tight junction status during chronic cerebral hypoperfusion.

The seed extract of Cassia obtusifolia ameliorates learning and memory impairments induced by scopolamine or transient cerebral hypoperfusion in mice.

In the present study, we assessed the effect of the ethanolic extract of the seeds of Cassia obtusifolia (COE) on the learning and memory impairments induced by scopolamine or transient bilateral common carotid artery occlusion (2VO). In a study of the cholinergic dysfunction induced by scopolamine, single COE (25, 50, or 100 mg/kg, p.o.) administration significantly attenuated scopolamine-induced cognitive impairments as determined by the passive avoidance and Y-maze tasks (P<0.05) and also reduced escape-latency on the Morris water maze task (P<0.05). In the 2VO study, COE (50 mg/kg, p.o.) significantly reversed 2VO-induced cognitive impairments in mice by the passive avoidance and the Y-maze tasks (P<0.05). Moreover, COE (50 mg/kg, p.o.) also reduced escape-latency and prolonged swimming time in the target quadrant during a probe trial of the Morris water maze task (P<0.05). In an in vitro study, COE was found to inhibit acetylcholinesterase activity in a dose-dependent manner (IC(50) value: 81.6 microg/ml). Furthermore, COE also inhibited acetylcholinesterase activity in an ex vivo study. These results suggest that COE attenuates memory impairment induced by scopolamine or 2VO and that these effects are mediated by enhancing the cholinergic nervous system via acetylcholinesterase inhibition.

Tanshinone congeners improve memory impairments induced by scopolamine on passive avoidance tasks in mice.

Tanshinones are a group of diterpenoids found in the roots of Salvia miltiorrhiza Bunge which has been used to treat cardiac disease. In the present study, we investigated the effect of the tanshinone congeners, tanshinone I, tanshinone IIA, cryptotanshinone, and 15, 16-dihydrotanshinone I, on learning and memory impairments induced by scopolamine (1 mg/kg, i.p.), a muscarinic antagonist, using passive avoidance tasks in mice. Tacrine was used as a positive control. Tanshinone I (2 or 4 mg/kg, p.o.), tanshinone IIA (10 or 20 mg/kg, p.o.), cryptotanshinone (10 mg/kg, p.o.), and 15, 16-dihydrotanshinone I (2 or 4 mg/kg, p.o.) significantly reversed scopolamine-induced cognitive impairments (P<0.05). Tanshinone I (2 mg/kg, p.o.) and tanshinone IIA (10 or 20 mg/kg, p.o.) were also reversed diazepam-induced cognitive dysfunctions (P<0.05). In addition, cryptotanshinone and 15, 16-dihydrotanshinone I were found to have an inhibitory effect on acetylcholinesterase in vitro with IC(50) values 82 and 25 microM, respectively. Furthermore, cryptotanshinone inhibited acetylcholinesterase activity for 3 h and 15, 16-dihydrotanshinone I for 6 h in an ex-vivo study. These results suggest that tanshinone congeners may be useful for the treatment of cognitive impairment and that their beneficial effects are mediated, in part, by cholinergic signaling enhancement.

Schizandra chinensis and Scutellaria baicalensis counter stress behaviors in mice.

The individual and combined antistress effects of the fruit of Schizandra chinensis and the radix of Scutellaria baicalensis were evaluated using a mouse acute stress model. Stress consisted of immobilization and electric foot shocks over 5 days. Mice were treated with herbal extracts for 7 days before exposing the animals to stress. Before each stressor presentation, the mice were treated with each herbal extract. Reduced locomotor activity and the percentage of time spent in the open arms of an elevated plus-maze under stress were recovered by treatment with the extract containing equal amounts of S. chinensis and S. baicalensis (CB11) at 200 and 400 mg/kg (p < 0.05). The effects of CB11 were greater than the effects of S. chinensis or S. baicalensis alone. CB11 treatment (100, 200 and 400 mg/kg) significantly reduced serum corticosterone levels (p < 0.05). Spleen size and the serum interleukin-2 level decreases induced by stress were prevented by CB11 (200 mg/kg) (p < 0.05). Taken together, these results suggest that S. chinensis and S. baicalensis in equal amounts could be used to treat stress disorders, in part, by preventing corticosterone and IL-2 level changes and ameliorating stress-related behavior parameters.

The ameliorating effect of oroxylin A on scopolamine-induced memory impairment in mice.

Oroxylin A is a flavonoid and was originally isolated from the root of Scutellaria baicalensis Georgi., one of the most important medicinal herbs in traditional Chinese medicine. The aim of this study was to investigate the ameliorating effects of oroxylin A on memory impairment using the passive avoidance test, the Y-maze test, and the Morris water maze test in mice. Drug-induced amnesia was induced by administering scopolamine (1 mg/kg, i.p.) or diazepam (1 mg/kg, i.p.). Oroxylin A (5 mg/kg) significantly reversed cognitive impairments in mice by passive avoidance and the Y-maze testing (P<.05). Oroxylin A also improved escape latencies in training trials and increased swimming times and distances within the target zone of the Morris water maze (P<.05). Moreover, the ameliorating effects of oroxylin A were antagonized by both muscimol and diazepam (0.25 mg/kg, i.p., respectively), which are GABA(A) receptor agonists. Furthermore, oroxylin A (100 microM) was found to inhibit GABA-induced inward Cl(-) current in a single cortical neuron. These results suggest that oroxylin A may be useful for the treatment of cognitive impairments induced by cholinergic dysfunction via the GABAergic nervous system.

Effect of the flavonoid, oroxylin A, on transient cerebral hypoperfusion-induced memory impairment in mice.

Oroxylin A is a flavonoid compound that is found in the root of Scutellaria baicalensis Georgi. The aim of this study was to determine the effects of oroxylin A on memory impairment induced by transient bilateral common carotid artery occlusion (2VO) in mice. The ameliorating effect of oroxylin A on memory impairment was investigated using a passive avoidance task, the Y-maze task, and the Morris water maze task in mice. Oroxylin A was found to significantly reverse 2VO-induced cognitive impairments in the passive avoidance and Y-maze tasks in a dose dependant manner (P<0.05). Moreover, oroxylin A (5 mg/kg, p.o.) shortened the escape-latency and prolonged swimming times in the target quadrant during the probe trial in the Morris water maze task (P<0.05). Histochemical and immunohistochemical studies showed that the number of Nissl bodies and OX-42 positive cells in the hippocampal CA1 and dentate gyrus regions were attenuated by oroxylin A. Moreover, phosphorylated cAMP response element-binding protein (CREB) and brain derived neurotrophic factor (BDNF) positive cell numbers were markedly increased in animals treated with oroxylin A than in untreated 2VO controls. These results suggest that oroxylin A dramatically attenuates the memory impairment induced by 2VO, and that this effect may be mediated by the neuroprotective effects of oroxylin A as supported oroxylin A induced reductions in activated microglia and increases in BDNF expression and CREB phosphorylation.

Gomisin A improves scopolamine-induced memory impairment in mice.

Gomisin A is a component of the fruits of Schizandra chinesis which are widely used as a tonic in traditional Chinese medicine. In the present study, we assessed the effect of gomisin A on the learning and memory impairments induced by scopolamine. The cognition-enhancing effect of gomisin A was investigated using a passive avoidance test, the Y-maze test, and the Morris water maze test in mice. Drug-induced amnesia was induced by treating animals with scopolamine (1 mg/kg, i.p.). Gomisin A (5 mg/kg, p.o.) administration significantly reversed scopolamine-induced cognitive impairments in mice by the passive avoidance test and the Y-maze test (P<0.05), and also improved escape latency in the Morris water maze test at 5 mg/kg (P<0.05). Moreover, in an in vitro study, gomisin A was found to inhibit acetylcholinesterase activity in a dose-dependent manner (IC50 value; 15.5 microM). These results suggest that gomisin A may be a useful cognitive impairment treatment, and its beneficial effects are mediated, in part, via enhancing the cholinergic nervous system.

Inhibitory effects of volatile antioxidants found in various beans on malonaldehyde formation in horse blood plasma.

The inhibitory effect of aroma extracts isolated from dried soybeans, mung beans, kidney beans, and azuki beans on malonaldehyde (MA) formation from horse blood plasma oxidized with Fenton's reagent was determined by gas chromatography (GC) coupled with nitrogen-phosphorus detector (NPD). Aroma chemicals such as maltol, eugenol, benzyl alcohol, 1-octen-3-ol, butyrolactone, and 1-methyl-2-pyrrolidone, found in the aroma extracts of beans, were also examined for their inhibitory effect on the same system. Among the four aroma extracts tested, the aroma extract of soybeans exhibited the strongest antioxidant activity. Extracts of soybeans, mung beans, azuki beans, and kidney beans inhibited MA formation by 58%, 47%, 40%, and 23%, respectively, at the level of 400 microg/mL, whereas, alpha-tocopherol and BHT inhibited MA formation by 52% and 70%, respectively, at the same level. Among the tested aroma chemicals, the antioxidant activity decreased in the following order: eugenol>maltol>1-octen-3-ol>benzyl alcohol>butyrolactone>1-methyl-2-pyrrolidone.