Novel function of bovine milk-derived lactoferrin on antinociception mediated by mu-opioid receptor in the rat spinal cord

Lactoferrin (LF) is a multifunctional protein that is found in milk, neutrophils, and other biological fluids. Although LF and the LF receptor have been identified in the central nervous system (CNS), the physiological role of LF remains unknown. We found that bovine milk-derived LF (BLF) reduces nociception in various pain models, as shown by the formalin test, hot plate test, and acetic acid writhing test in rats. Intraperitoneal (i.p.) administration of BLF significantly inhibited nociception in these pain models. These antinociceptive effects were also confirmed in BLF-fed rats. The antinociceptive effects of BLF were blocked by naloxone treatment, even though prostaglandin E(2) (PGE(2)) production in the ascites fluid that accumulated during the writhing test was not affected by BLF. Intrathecal (i.t.) application of BLF caused marked antinociceptive effects that were reversed by co-administration of a specific mu-opioid receptor antagonist, D-Phe-Cys-Tyr-D-Trp-Orn-Thr-NH(2) (CTOP), or by naloxone during the formalin test. We conclude that LF possesses mu-opioid receptor-mediated antinociceptive activity in the spinal cord.     

Maternal stress differently alters nociceptive behaviors in the formalin test in adult female and male rats

The long-term effects of restraint stress in Wistar rats during the last week of gestation were investigated on the acute and tonic phases of the specific biphasic nociceptive behavioral response in the formalin test in offspring, females and males, at 90 days. Prenatal stress produced significant changes in formalin-induced pain, which was more pronounced in females as compared to males. The distorted response in females was more at the supraspinal level with an increased intensity of the licking response in both phases as well as with an increased their duration. Results concerning changes of the interphase length indicate the impairments of inhibitory mechanisms in the central nervous system. Furthermore, profound difference in the effects of prenatal stress on the first phase but similarity in these effects on the second phase in females and males are indirect strong support of the view that the second phase in the formalin test can not be mediated by central sensitization alone but greatly depends on signals ongoing from nociceptive primary afferents. Finally, the results obtained in males are important argument in favor of assumption about different mechanisms of acute and tonic pain. Taken together, these studies show that prenatal stress alters nociceptive behaviors in the formalin test in rats at 90 days in a sex-specific manner.     

Chronic psychosocial stress impairs learning and memory and increases sensitivity to yohimbine in adult rats

BACKGROUND: It is well known that intense and prolonged stress can produce cognitive impairments and hippocampal damage and increase noradrenergic activity in humans. This study investigated the hypothesis that chronic psychosocial stress would affect behavior, drug sensitivity, and hippocampal-dependent learning and memory in rats. The work provides a novel connection between animal and human studies by evaluating the effects of stress on a rat's response to yohimbine, an alpha(2) adrenergic receptor antagonist. METHODS: Rats were exposed to a cat for 5 weeks and randomly housed with a different group of cohorts each day (psychosocial stress). The effects of the stress manipulations were then assessed on open field behavior, spatial learning and memory in the radial arm water maze and the behavioral response to a low dose of yohimbine (1.5 mg/kg). RESULTS: Stressed rats displayed impaired habituation to a novel environment, heightened anxiety, and increased sensitivity to yohimbine. In addition, the stressed rats exhibited impaired learning and memory. CONCLUSIONS: There are commonalities between the current findings on stressed rats and from studies on traumatized people. Thus, psychosocial stress manipulations in rats may yield insight into the basis of cognitive and neuroendocrine disturbances that commonly occur in people with anxiety disorders.     

Differential effects of psychological stress on activation of the 5-hydroxytryptamine- and dopamine-containing neurons in the brain of freely moving rats

We investigated the effects of psychological stress, lacking direct physical stimulus, on the release of 5-hydroxytryptamine (5-HT) and dopamine (DA) in the basolateral nucleus of the amygdala (BLA) and the dorsal raphe nuclei (DRN) in the rat using the in vivo microdialysis technique with dual probes, one in each region of the same animals. Psychological stress was employed using the communication box paradigm. Psychological stress for 1 h significantly increased dialysate 5-HT levels in the DRN and the BLA. Psychological stress-induced 5-HT release in the BLA was significantly greater than those in the DRN, indicating that modifications of the serotonergic neurons in the BLA are more sensitive to psychological stress than are those of the DRN. Psychological stress also increased DA release in the BLA, while the dialysate DA levels in the DRN were unchanged. These results suggest that psychological stress preferentially activates ascending serotonergic neurons from the DRN to the BLA but not those of dopaminergic neurons. Furthermore, our findings indicate that both the serotonergic neurons and the dopaminergic neurons in the BLA may have a distinct role to play in the neuronal responses to psychological stress.     

大鼠实验性脑损伤血浆中NO及NOS的动态变化研究

目的 测定实验性脑损伤后血浆中一氧化氮含量、一氧化氮合酶活性,研究其与脑水肿之间的关系.方法 大鼠随机分组,用硝酸还原酶法测定血清中NO含量、NOS活性,及测定脑组织含水量.并行还原型辅酶Ⅱ依赖性黄递酶(NADPⅡ-d)组化染色检测皮层及脑底NOS阳性细胞.结果 (1)TBI后血浆内NO含量、NOS活力即有升高,与对照组比较有明显差异(P＜0.05).(2)脑组织含水量在外伤后升高,与对照组比较有明显差异(P＜0.05).与血浆中NO含量、NOS活力变化趋势一致.(3) NADPⅡ-d组化染色显示TBI皮层NOS阳性细胞明显多于正常对照组,伤灶脑底也出现了染色块及浓染的细胞群与阳性纤维束.结论 大鼠TBI后NO含量、NOS活性的升高,与脑水肿的发生有关.     

Effects of anxiety on TNF-alpha levels during psychological stress

OBJECTIVES: Psychological stress can influence the immune system, which may result in stress-related illnesses. In this study, we investigated the effect of psychological stress and the coping skill on plasma cytokine levels. MATERIALS AND METHODS: One hundred eighty-three students, at different stages of an academic year, participated in this study. Plasma tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-2 soluble receptor alpha, and IL-4 were measured and examined in relation to the measures of anxiety [State Anxiety Inventory (SAI)] and Bell Adjustment Inventory (BAI) score. RESULTS: SAI scores were significantly higher in both midterm students (MTS) and examination-taking students (ETS), compared with the freshly admitted students (FAS). In addition, TNF-alpha levels were significantly different between the high- and the low-anxiety groups of ETS but not in MTS or FAS. The correlation between SAI scores and the BAI emotional scores was highest in the ETS group. CONCLUSIONS: TNF-alpha level was significantly lower in the ETS group with high anxiety scores, and it is situation specific.     

Adrenocortical suppression blocks the enhancement of memory storage produced by exposure to psychological stress in rats

Several reports have indicated that various stress stimuli modulate learning and memory processes. In the present study, the effects of adrenocortical suppression with the 11beta-hydroxylase inhibitor metyrapone on the psychological stress-induced changes in memory storage in inhibitory avoidance training and in serotonin turnover in various brain regions were investigated in rats. Retention of one-trial inhibitory avoidance and the plasma corticosterone level were significantly enhanced by post-training exposure to psychological stress for 1 h. Pretreatment with metyrapone (12.5 or 25 mg/kg, s.c.) 90 min beforehand dose-dependently blocked the enhancement of memory storage and of the plasma corticosterone level produced by psychological stress. These results suggest that the adrenocortical system may contribute to the memory-enhancing effect of psychological stress. In a neurochemical study, a significant increase in serotonin turnover in the hippocampus and limbic forebrain, including the nucleus accumbens, were observed in rats that were exposed to psychological stress. In contrast to the behavioral experiments, these changes in serotonin turnover produced by exposure to psychological stress were not antagonized by pretreatment with metyrapone; instead, a further increase in serotonin turnover was observed only in the hippocampus. These results suggest that the serotonergic system in the hippocampus might be selectively regulated by adrenal steroids in response to stress, and imply the existence of negative feedback mechanisms via a hippocampal serotonergic system in the memory enhancement associated with corticosterone and psychological stress.     

一氧化氮及一氧化氮合酶抑制剂在大鼠局灶性脑缺血时作用研究

目的：观察一氧化氮及一氧化氮合酶抑制剂在大鼠局灶性脑缺血时的作用方法;民凝大鼠大脑中动脉制成脑缺血模型,选择脑缺血３０ｍｉｎ,６０ｍｉｎ,１２０ｍｉｎ,１８０ｍｉｎ为研究时点,观察各时点用选择性,非选择性一氧化氮合酶抑制剂亚硝酸盐含量测定,缺血脑组织坏死体积测定及损伤海马ＣＡ１电镜观察。     

Inducible nitric oxide synthase expression is selectively induced in astrocytes isolated from adult human brain

Inducible nitric oxide synthase (iNOS) expression has been shown to be differentially regulated among different cell types and species. In cultures of primary human fetal glial cells, we have shown that astrocytes rather than microglia express iNOS. In the present study, we extended these findings to primary cultures of astrocytes and microglia derived from adult human brains. Mixed cultures of adult brain tissue were stimulated with IL-1β and IFNγ, a combination known to induce iNOS maximally in human fetal cells, and the expression of iNOS was determined by immunocytochemistry. Cell types were determined by morphology as well as immunocytochemistry for GFAP (astrocytes) and CD68 (microglia). The results showed that in cultures of adult human glia, iNOS was expressed following stimulation with cytokines, and the expression was restricted to astrocytes. Astrocyte iNOS immunoreactivity was detected both in the cytosol and in a discrete paranuclear region, a pattern noted in human fetal astrocytes. These results demonstrate that the ability to express iNOS is common to both fetal and adult human astrocytes.     

In the search for integrative biomarker of resilience to psychological stress

Psychological resilience can be defined as individual’s ability to withstand and adapt to adverse and traumatic events. Resilience is traditionally assessed by subjective reports, a method that is susceptible to self-report bias. An ideal solution to this challenge is the introduction of standardised and validated physiological and/or biological predictors of resilience. We provide a summary of the major concepts in the field of resilience followed by a detailed critical review of the literature around physiological, neurochemical and immune markers of resilience. We conclude that in future experimental protocols, biological markers of resilience should be assesses both during baseline and during laboratory stressors. In the former case the most promising candidates are represented by heart rate variability and by in vitro immune cells assay; in the latter case—by startle responses (especially their habituation) during stress challenge and by cardiovascular recovery after stress, and by cortisol, DHEA and cytokine responses. Importantly, they should be used in combination to enhance predictive power.     

青少期社会隔离和不确定性应激对大鼠成年后情绪行为的影响

目的 分别观察青少期社会隔离和不确定性应激对大鼠成年后情绪行为的影响.方法在青少期(出生后28 d～41 d)分别给予研究组大鼠2周社会隔离应激或不确定性复合应激,应激后经3周恢复至所有大鼠进入成年期后检测其情绪行为改变.采用糖水偏好测试和强迫游泳检测大鼠的抑郁样行为,采用旷场试验和高架十字迷宫试验检测焦虑样行为.结果 与群养组相比,青少期社会隔离组大鼠成年后的所检测的各项情绪行为均无明显差异(P>0.05).经历青少期不确定性应激的成年大鼠与其对照组相比糖水偏好指数没有明显改变[(0.86±0.19) vs (0.86±0.03),P>0.05],但在强迫游泳测试中表现出更多的绝望行为(强迫游泳不动行为增加)[(29.88±3.37) vs (19.90±3.19),P<0.05]、更少的主动行为(挣扎行为减少)[(24.00±1.67) vs (32.90±3.09),P<0.05],提示该应激并不影响大鼠成年后偏好糖水的本能行为,但导致其急性应激应对能力受损,对成年应激事件更容易发展出次级的应对无能或绝望的抑郁样行为.不确定性应激研究组的大鼠成年后在高架十字迷宫测试中较其对照组闭合臂停留时间明显增加[(176.90±17.01) vs (136.48±9.47),P<0.05]以及开放臂进入次数明显减少[(3.00±0.93) vs (5.90±1.08),P<0.05],提示该应激增加了大鼠成年后的焦虑样行为水平.结论 青少期社会隔离应激和不确定性应激对大鼠成年后情绪行为的影响存在差异,青少期不确定性应激能够有效诱导大鼠多项情绪行为显著和持续的改变,提供了一种青少期应激增加成年大鼠应激性抑郁易感性的动物模型.     

Effects of shear stress on nitric oxide levels of human cerebral endothelial cells cultured in an artificial capillary system

Nitric oxide (NO) is an important vasodilator with various activities in the cerebral vasculature. Although the response of NO levels to shear stress has been investigated in various models using systemic endothelium, no study has evaluated human cerebral endothelial cells (HCE). We determined the NO levels of HCE cultured in an artificial capillary system in response to changes in shear stress. With direct measurement by a porphyrinic microsensor, we found that NO levels increased immediately with a peak at 7 h after changes in shear stress, and by 24 h dropped to a constant elevated baseline. Shear stress-mediated increases in NO levels were confirmed by the measurement of citrulline, an indirect measure of NO. Furthermore, NO levels by HCE were shown to decrease with decreasing shear stress levels. This study presents a novel system to study NO production by microvascular HCE, and indicates a linear relationship between shear stress and NO levels. As cerebral vessels age and lose transmural compliance, shear stress-mediated production of NO may play a greater role in cerebrovascular function and dysfunction.     

七叶皂苷钠对大鼠脑缺血再灌注损伤的保护作用

目的 探讨七叶皂苷钠对大鼠脑缺血再灌注损伤的作用及机制.方法 60只成年SD大鼠随机分为假手术组、模型组和七叶皂苷钠组,各20只.采用线栓法建立大鼠局灶性大脑中动脉闭塞再灌注损伤模型,七叶皂苷钠组建模成功后1h腹腔注射七叶皂苷钠(2.8 mg/kg),假手术组和模型组腹腔注射等量生理盐水.造模后24 h,使用Longa...     

Oxidative stress in prefrontal cortex of rat exposed to MK-801 and protective effects of CAPE

MK-801 was shown to be one of the most neurotoxic non-competitive NMDA receptor antagonists. It is known that repeated injection of MK-801 was proposed in an animal model in psychosis. The aims of this study are to investigate the contributing effect of oxidative stress in MK-801-induced experimental psychosis model, and to show that prevention of oxidative stress may improve prognosis. Furthermore, there is evidence that oxygen free radicals play an important role in the pathophysiology of schizophrenia. In this study, Wistar Albino rats were divided into three groups: 1st group: Control, 2nd group: MK-801, 3rd group: MK-801+CAPE (Caffeic acid phenethyl ester) group. MK-801 was given intraperitoneally at the dose of 0.5 mg/kg/day for 5 days. CAPE was given to the treatment group while exposed to MK-801. In control group, saline was given intraperitoneally at the same time. After 7 days, rats were killed by decapitation. Prefrontal cortex (PFC) of rats was removed for biochemical and histological analyses. As a result, malondialdehyde (MDA), protein carbonyl (PC), nitric oxide (NO) levels and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and xanthine oxidase (XO) and adenosine deaminase (AD) enzyme activities were found to be increased significantly in prefrontal cortex (PFC) of MK-801 group (p<0.0001) compared to control group. In CAPE treated rats, prefrontal tissue MDA, PC, NO levels and, GSH-Px, XO, AD enzyme activities were significantly decreased when compared to MK-801 groups (p<0.0001) whereas catalase (CAT) enzyme activity was not changed. Moreover, in the light of microscopic examination of MK-801 groups, a great number of apoptotic cells were observed. CAPE treatment decreased the apoptotic cell count in PFC. The results of this study showed that MK-801-induced neurotoxicity caused oxidative stress in PFC of rats. This experimental study may also provide some evidences for the new treatment strategies with antioxidants in schizophrenia.     

Aproteic diet decreases hypothalamic catecholamine turnover in adult male rats

Previous reports indicate that malnutrition reduces reproductive functions. We have demonstrated that protein deprivation in the diet also causes reproductive dysfunction by reducing hypothalamic GnRH secretion. Noradrenaline and nitric oxide are modulators of GnRH secretion. Noradrenaline stimulates GnRH secretion and nitric oxide inhibits catecholamine release. This work studies the hypothalamic catecholaminergic and nitrergic neuron activity in Wistar adult male rats fed on an aproteic diet (AP) during 21 days; this treatment was started when rats were 70 days old. Our first experiment studied catecholamine turnover rate after inhibition of tyrosine hydroxylase activity by injecting (i.p.) 400 mg/kg alpha-methyl-p-tyrosine. Our second experiment studied in vitro hypothalamic nitric oxide synthase (NOS) activity in animals under the same diet. AP diet significantly decreased both noradrenaline (P<0.05) and dopamine (P<0.05) hypothalamic turnover rate. Noradrenaline turnover in cerebral cortex was not altered by the aproteic diet. However, hypothalamic NOS activity was not affected in animals fed on an AP diet. These results indicate that the lack of protein in the diet reduces catecholaminergic neuron activity in adult male rats by a NO-independent mechanism, thus suggesting that a decrease in noradrenergic activity may be involved in the reduction of GnRH secretion induced by an AP diet.     

Immobilization-induced stress activates neuronal nitric oxide synthase (nNOS) mRNA and protein in hypothalamic-pituitary-adrenal axis in rats

The purpose of this study was to determine whether immobilization stress can cause changes in the enzyme activity and gene expression of neuronal nitric oxide synthase (nNOS) in the hypothalamus, pituitary, and adrenal gland in rats. NOS enzyme activity was measured as the rate of [³H]arginine conversion to citrulline, and the level of nNOS mRNA signal was determined using in situ hybridization and image analysis. NOS-positive cells were also visualized using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase) histochemistry and by immunohistochemistry using an anti-nNOS antibody. A significant increase of NOS enzyme activity in the anterior pituitary, adrenal cortex, and adrenal medulla (1.5-, 3.5-, and 2.5-fold) was observed in the stressed animals (immobilization of 6 h) as compared to non-stressed control rats. Up-regulation of nNOS mRNA expression in anterior pituitary and adrenal cortex was already detectable after stress for 2 h with 1.5- and 2-fold increase, respectively. The nNOS mRNA signals in hypothalamic paraventricular nucleus (PVN) significantly increased after the stress for 6 h. This increase in NOS enzyme activity was confirmed using NADPH-diaphorase staining and immunostaining in the PVN and adrenal cortex. An increase of NOS enzyme activity in adrenal medulla after immobilization for 6 h posited by far longer than in the adrenal cortex and anterior pituitary. The present findings suggest that psychological and/or physiological stress causes NO release in hypothalamic-pituitary-adrenal (HPA) axis and in sympatho-adrenal system. It is suggested that NO may modulate a stress-induced activation of the HPA axis and the sympatho-adrenal medullary system. The different duration of stress-induced NOS activity in HPA axis and the adrenal medulla may suggest NO synthesis is controlled by separate mechanism in the two HPA and the sympatho-adrenal systems.     

Investigation into the cross-correlation of salivary cortisol and alpha-amylase responses to psychological stress

Stress is a multidimensional construct. To accurately represent stress physiology, multiple stress measures across multiple stress-related systems should be assessed. However, associations may be masked given that different systems underlie different time courses. Salivary cortisol and alpha-amylase (sAA) are reliable biological stress markers of the sympathetic nervous system (SNS) and the hypothalamus pituitary adrenal (HPA) axis, respectively. Studies examining the link between sAA and cortisol levels in response to stress have produced inconsistent results. Here, we investigated whether the covariance of stress-induced sAA and cortisol release is dependent on the distinct temporal dynamics of the two stress markers. A total of 50 male participants were exposed to a psychological laboratory stressor with high frequency (2-min interval) saliva sampling in two independent studies. Synchronized time series of sAA and cortisol measures before, during and after stress induction were obtained. Cross-correlation analysis was applied to test for the association of sAA and cortisol levels at various stages relative to the onset of the stressor. Positive and negative cross-correlations between lagged pairs of sAA and cortisol measures were found in both studies. The strongest correlation was found for sAA preceding cortisol release by 13.5 min (r = .27, p < .001). With a smaller effect size cortisol also significantly preceded sAA by 13.5 min (r = -.16, p < .001). We suggest that sAA and cortisol stress responses are reliably associated at various time lags throughout a stressful situation. As a possible connection site between HPA axis and SNS that may underlie sAA-cortisol associations, we discuss CRF neurons of the hypothalamus involved in sympathetic regulation.     

Synaptic plasticity to estrogen in the lateral septum of the adult male and female rats

There is no sex difference in the number of axodendritic synapses in the lateral septum of adult rats. However, the treatment of adult females with estradiol (E2) for 4 weeks increases significantly the number of synapses, whereas E2 treatment fails to increase the synaptic number in males, suggesting a possible sexually dimorphic synaptic response to estrogen.     

Relationship between brain zinc and transient learning impairment of adult rats fed zinc-deficient diet

Because of the reported presence of both CART peptide and NOS activity in the same hypothalamic nuclei, their colocalization was examined. Eighteen percent of the neurons in the supraoptic nuclei, and 16% of the neurons in the paraventricular nucleus contained both CART immunoreactivity and NOS activity. Many other neurons in these regions stained for only one marker although they were often close by. Thus, CART peptides and NO may interact in these regions.