Antioxidant flavonoid glycosides from Evolvulus alsinoides

Oxidative damage is an established outcome of chronic stress. Thus, the present study was designed to investigate the modulatory role of ethanolic extract of Evolvulus alsinoides (EA) in terms of oxidative alterations at peripheral and central level in rats subjected to chronic unpredictable stress (CUS). CUS exposure for 7 days reduced Cu, Zn superoxide dismutase and catalase activity with increase in glutathione peroxidase activity and lipid peroxidation, while decrease in reduced glutathione level in blood plasma, frontal cortex and hippocampus regions of brain. Oral administration of EA extract at 200 mg/kg p.o. normalized these stress induced oxidative alterations with an efficacy similar to that of melatonin. Further, EA extract was taken up for detailed chemical investigation. Two new flavonol-4′-glycoside, kaempferol 4′-O-β-d-glucopyranosyl-(1 → 2)-β-d-glucopyranoside (3) and kaempferol 4′-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside (5) were isolated, along with eight known compounds (1, 2, 4 and 6–10). The structures of new compounds were established by detailed spectroscopic studies, while known compounds were characterized by direct comparison of their reported NMR data. All these compounds were evaluated for their in vitro antioxidant activity. Compounds 3, 5, 9 and 10 at 100 and 200 μg/ml showed significant in vitro antioxidant activity. Therefore, EA may hold great potential in preventing clinical deterioration in stress induced oxidative load and related disorders.     

Prey–predator dynamics in seahorses (Hippocampus guttulatus): deciphering fatty acid clues

Understanding the flow of fatty acids between trophic levels can provide important clues on prey–predator dynamics and nutritional requirements of the species. This study investigates the fatty acid flow between enrichment emulsions, Artemia nauplii and Hippocampus guttulatus juveniles, and evaluates the nutritional value of enriched and unenriched Artemia for newborn seahorses. The fatty acid profile of Artemia and seahorses generally reflected the dietary composition, but fatty acids were not linearly transferred between trophic levels. The incorporation of dietary fatty acids showed to be a more complex process involving dietary composition, predator metabolism and nutritional requirements. Artemia composition resulted from a dynamic balance between what was assimilated and metabolized by the nauplii during enrichment. Prey fatty acids were incorporated in seahorses, but HUFA, particularly DHA, were selectively retained to fulfil their high requirements. H. guttulatus newborns were not successfully reared on Artemia nauplii, not even on enriched Artemia, with low survival rates (15.0–26.7%) being observed in all feeding treatments. The high MUFA content and low DHA level of Artemia did not fulfil the high SFA and PUFA requirements of newborn juveniles, particularly their great DHA demands. Higher survivorship was obtained with enriched Artemia, but no differences were detected in juvenile growth.     

Absence of temporal lobe epilepsy pathology in dogs with medically intractable epilepsy

Epilepsy is a common neurological problem in dogs. In some dogs, seizures cannot be controlled adequately with anticonvulsant medication. Temporal lobe epilepsy is the most common type of epilepsy in adult humans, it is frequently resistant to anticonvulsant therapy, and it is commonly associated with characteristic neuropathological abnormalities in the hippocampal dentate gyrus. We sought to test the hypothesis that dogs with medically intractable epilepsy have temporal lobe epilepsy. The hippocampi of 6 dogs that were euthanized because of chronic, recurrent seizures were compared with those of 8 nonepileptic controls. In control and epileptic dogs, stereological cell counting showed similar numbers of neurons in the hilus of the dentate gyrus, somatostatin immunoreactivity identified numerous immunopositive neurons in the hilus, and Timm staining showed the normal pattern of granule cell axon projections. These findings demonstrate a lack of hilar neuron loss and granule cell axon reorganization, suggesting that temporal lobe epilepsy is not a common cause of medically intractable epilepsy in dogs.     

Postconditioning modulates TLR4 expression in hippocampus of tree shrews with thrombotic cerebral ischemia

AIM: To observe the effects of thrombotic cerebral ischemia and postconditioning on the expression of toll-like receptor 4 (TLR4) in hippocampus of tree shrews.METHODS: The model of thrombotic focal cerebral ischemia was established by photochemical reaction.Four hours after the onset of photochemical reaction, ischemic postconditioning was induced by 3 repeated cycles of carotid artery occlusion for 5 min and reperfusion for 5 min. The histological changes of hippocampus (by HE staining), TLR4 protein level (by Western blotting) and TLR4 mRNA expression (by semiquantitative RT-PCR) were observed.RESULTS: The extensive neuronal degeneration in hippocampus was observed from 4 h to 72 h and peaked at 24 h after cerebral ischemia, but was significantly attenuated after postconditioning. Cerebral ischemia caused a progressive increase in the expression of TLR4 protein at 4 h and 24 h (P<0.05), and decreased at 72 h (P<0.05). In contrast to ischemia groups, postconditioning decreased the expression of TLR4 protein at 4 h and 24 h (P<0.05), but an increase in the expression of TLR4 at 72 h (P<0.05) was observed. Simultaneously, the level of TLR4 mRNA in hippocampus showed the tendency of approximate variation in accordance with the protein expression.CONCLUSION: The expression of TLR4 increases by cerebral ischemia. The protection mechanisms of postconditioning may be associated with modulating TLR4 expression.     

海马多糖提取工艺优化与抗氧化活性研究

以日本海马（Hippocampus mohnikei）为原料,采用响应面试验优化超声辅助提取粗多糖工艺条件,通过柱层析进行纯化,测定海马多糖的单糖组成,并评价其抗氧化活性。结果表明：超声时间47 min、浸提温度87 ℃、液料比23 mL/g为最佳提取条件,粗多糖HP得率为9.91%;通过柱色谱分级纯化得到2种多糖HP-1和HP-2,HP-1主要由葡萄糖、半乳糖、甘露糖以摩尔比11.32: 6.39: 5.64组成,并含有少量阿拉伯糖、木糖、岩藻糖、鼠李糖,HP-2主要由氨基葡萄糖、葡萄糖醛酸、半乳糖醛酸、半乳糖、葡萄糖、甘露糖以摩尔比 29.03: 18.18: 8.48: 6.30: 4.88: 2.70组成,并含有少量岩藻糖和木糖;HP-1、HP-2具有良好DPPH自由基清除能力,且HP-2最强,同时HP、HP-1及HP-2均具有还原力,表明海马多糖具有良好抗氧化活性。     

Phosphodiesterase 4 inhibitor rolipram prevents chronic alcoholism and withdrawal-induced depression-like behaviors in mice

AIM: To investigate the effect of phosphodiesterase 4 (PDE4) inhibitor rolipram on the levels of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP response element-binding protein (CREB), phosphorylated CREB (p-CREB) and brain-derived neurotrophic factor (BDNF) in the hippocampus and the prefrontal cortex (PFC) of alcoholism model mice.METHODS: The mice (n=60) were randomly divided into control group, control+rolipram group, alcoholism model group, and alcohol+rolipram (0.1, 0.5 and 1 mg/kg) groups. The mice were given alcohol preference test on days 6, 13, 20 and 27. After the test, the mice received withdrawal of alcohol for 1 d. On day 28, the mice were given behavior test of depression, and after the test, the mice were sacrificed. The cAMP levels in the hippocampus and PFC were detected by ELISA, and the protein levels of PKA, CREB, p-CREB and BDNF were detected by Western blot.RESULTS: The mice showed an obvious drinking phenomenon (P<0.01), and the immobility time of forced swimming test and tail suspension test was significantly increased (P<0.01), with increasing drinking days and withdrawal times. However, chronic treatment with rolipram for 28 d reversed this phenomenon. Moreover, the cAMP levels in the hippocampus and PFC were significantly decreased after 28 d alcohol treatment (P<0.01), and pretreatment with rolipram (1 mg/kg) obviously reversed this decrease (P<0.01). Parallel to these changes of cAMP, the protein levels of PKA, p-CREB and BDNF were also decreased in the hippocampus and PFC (P<0.01), and 28 d rolipram administration inhibited the decreased cAMP, PKA, p-CREB and BDNF levels in the hippocampus. Moreover, 28 d rolipram administration also reversed decreased cAMP, PKA and p-CREB in the PFC.CONCLUSION: Rolipram treatment protects against alcohol-induced depression-like behaviors, and also reduces alcohol drinking. These effects may be related to PDE4-cAMP-PKA-CREB-BDNF pathway.     

Effects of astragalan on neurotransmitters and expression of c-fos mRNA in hippocampus after ischemic brain injury in rats

AIM: To explore the effects of astragalan (AG) on the neurotransmitters,acetylcholine (ACh), norepinephrine (NE) and 5-hydroxytryptamine (5-HT), and the expression of c-fos mRNA in hippocampus after ischemic brain injury in rats. METHODS: Male Wistar rats (180~220 g) were randomly divided into 10 groups (n=10): sham-operated group (SOG), 3 model groups (MG 1 d, 3 d, 7 d) and 3 low- or high-dose AG treatment groups (L/H-AGTG 1 d, 3 d, 7 d), respectively. The middle cerebral artery of the rats in MG group and AGTG group were blocked by operation to induced brain injury. The cerebral blood vessels of the animals were blocked on day 1, day 2 and day 7, respectively, after the L/H-AGTG were treated with AG (5 mg/kg and 15 mg/kg, ip). The content of ACh,5-HT and NE was determined using their respective ELISA kits, and the expression of c-fos mRNA in the hippocampus homogenate was semiquantitative analyzed by RT-PCR after neurologic impairment (NIP) was scored. RESULTS: AG attenuated the injury in hippocampus by cerebral ischemia in a dose-dependent manner. The content of ACh, 5-HT and NE in L-AGTG 7 d,H-AGTG 3 d and 7 d groups was significantly higher than that in MG group, but was lower in SOG group (P<0.05 or P<0.01). The mRNA expression of c-fos in SOG group was lower than that in MG group (P<0.05 or P<0.01), indicating that reinforcement expression of c-fos mRNA by cerebral ischemia and the expression of downstream genes may be beneficial for protecting the neurons. The mRNA expression of c-fos in H-AGTG 3 d/7 d groups was higher than that in MG group (P<0.05). CONCLUSION: AG attenuates the damage of neurons and improves the functions of hippocampus under the condition of cerebral ischemia/reperfusion by increasing the content of ACh, NA and 5-HT, and the mRNA expression of c-fos in hippocampus.     

铜暴露对小鼠铜代谢相关酶表达及氧化应激的影响

[目的]研究长期低剂量铜暴露对小鼠海马组织铜含量、铜代谢相关酶及氧化应激的影响,为深入研究低剂量铜暴露的神经毒性作用提供基础数据。[方法]选择野生型小鼠,随机分为对照组和试验组对照组小鼠饮用蒸馏水,试验组在饮用水中添加0.13 mg/L氯化铜,连续饮用3个月,研究长期低剂量铜暴露对小鼠海马组织中铜含量、铜代谢相关酶表达及氧化应激的影响。[结果]与对照组相比,试验组小鼠海马中的自由铜及结合铜含量均显著升高,而铁、锌、钙离子含量在海马中均无显著差异;与对照组相比,试验组小鼠海马组织中铜蓝结合蛋白表达显著降低,蛋白硝基酪氨酸水平显著升高,8-羟基脱氧鸟嘌呤荧光表达水平显著升高。[结论]长期超低剂量铜暴露可导致铜在脑组织中聚集,但不影响铁、锌、钙和镁的含量;长期低剂量铜暴露可导致氧化应激。     

池养大海马的摄食_生长和生态转换效率

对大海马 (平均体长 92 .82± 2 .0 5 2 6mm ,平均体重 6 .6 9± 0 .4 5 70g)在养殖条件下 (放养密度为 2 0 0ind·m-3 ,溶解氧保持在 5 .5mg·L-1以上 ,生物耗氧量低于 3mg·L-1,pH值 7.5～ 8.2 ,透明度 5 0～ 85cm ,盐度 15 .2～ 2 6 .5 )的摄食、生长和食物生态转换效率进行了定量研究。通过系统取样 ,测量海马的全长、体重和消化道内食物重量 ,计算其增长速度、摄食率、消化道排空率、生态转换效率等数值 ,发现大海马的摄食每天有两个摄食高峰 ,即 12 :0 0和 18:0 0 ,晚上不摄食或很少摄食。大海马的日摄食量为 16 .6 342± 0 .782 0g·(10 0g) -1,日摄食率为 1177.2 5cal·ind-1,排空率 0 .14 4 4g·(10 0g) -1·h-1,食物转换效率 2 0 .0 4 % ,能量转换效率 31.4 2 %。体长日平均增长速度和日均增长率分别为 1.176 6mm和 1.2 7% ;体重日平均增重速度和日均增重率分别为0 .182 0g和 2 .2 7%。通过多元回归分析建立摄食率与体重及温度的相关关系 ,得出下列关系式 :C =6 .75 4 4+0 .30 14T +0 .2 190W (F =83.5 96 3,F0 .0 1=18.0 0 ,F >F0 .0 1)。