Role of Cannabinoid CB1 Receptor in Object Recognition Memory Impairment in Chronically Rapid Eye Movement Sleep-deprived Rats

Objective We aimed to investigate whether antagonism of the cannabinoid CB1 receptor(CB1R)could affect novel object recognition(NOR)memory in chronically rapid eye movement sleep-deprived(RSD)rats.Methods The animals were examined for recognition memory following a 7-day chronic partial RSD paradigm using the multiple platform technique.The CB1R antagonist rimonabant(1 or 3 mg/kg,i.p.)was administered either at one hour prior to the sample phase for acquisition,or immediately after the sample phase for consolidation,or at one hour before the test phase for retrieval of NOR memory.For the reconsolidation task,rimonabant was administered immediately after the second sample phase.Results The RSD episode impaired acquisition,consolidation,and retrieval,but it did not affect the reconsolidation of NOR memory.Rimonabant administration did not affect acquisition,consolidation,and reconsolidation;however,it attenuated impairment of the retrieval of NOR memory induced by chronic RSD.Conclusions These findings,along with our previous report,would seem to suggest that RSD may affect different phases of recognition memory based on its duration.Importantly,it seems that the CB1R may,at least in part,be involved in the adverse effects of chronic RSD on the retrieval,but not in the acquisition,consolidation,and reconsolidation,of NOR memory.

Novel Method for Synthesis of Diarylpyrazole Derivatives as Cannabinoid CB_1 Receptor Antagonists

A novel and efficient method was developed for the synthesis of diarylpyrazole derivatives as cannabinoid CB1 receptor antagonist via four step reactions. The key step was the synthesis of a diarylpyrazole skeleton, which involved initial condensation of the sodium salt of compound 12 with diazonium compounds, and further cyclization by heating at reflux in acetic acid. Eight diarylpyrazole derivatives and nine new synthesized compounds were characterized by 1H NMR, IR, MS, and elemental analysis. The reaction conditions were mild and the overall yields of the target compounds ranged from 26% to 44%.

AB059.Expression patterns of CB1R,NAPE-PLD,and FAAH in the primary visual cortex of vervet monkeys

Background:The expression,localization,and function of the endocannabinoid system has been well characterized in recent years in the monkey retina and in the primary thalamic relay,the lateral geniculate nucleus(dLGN).Few data are available on cortical recipients’structures of the dLGN,namely the primary visual cortex(V1).The goal of this study is to characterize the expression and localization of the metabotropic cannabinoid receptor type 1(CB1R),the synthesizing enzyme N-acyl phosphatidyl-ethanolamine phospholipase D(NAPE-PLD),and the degradation enzyme fatty acid amide hydrolase(FAAH)in the vervet monkey area V1.Methods:Using Western blots and immunohistochemistry,we investigated the expression patterns of CB1R,NAPE-PLD,and FAAH in the vervet monkey primary visual cortex.Results:CB1R,NAPE-PLD,and FAAH were expressed in the primary visual cortex throughout the rostro-caudal axis.CB1R showed very low levels of staining in cortical layer 4,with higher expressions in all other cortical layers,especially layer 1.NAPE-PLD and FAAH expressions were highest in layers 1,2 and 3,and lowest in layer 4.Conclusions:Interestingly enough,CB1R was very low in layer 4 of V1 in comparison to the other cortical layers.The visual information coming from the dLGN and entering layer 4Calpha(magno cells)and 4Cbeta(parvo cells)may be therefore modulated by the higher expression levels of CB1R in cortical layers 2 and 3 on the way to the dorsal and ventral visual streams.This is further supported by the higher expression of NAPE-PLD and FAAH in the outer cortical layers.These data indicate that CB1R system can influence the network of activity patterns in the visual stream after the visual information has reached area V1.These novel results provide insights for understanding the role of the endocannabinoids in the modulation of cortical visual inputs,and hence,visual perception.

AB007. Expression and localization of CB1R,NAPE-PLD,and FAAH in the primary visual cortex of vervet monkeys

Background:The goal of this study is to determine the expression and localization of the cannabinoid receptor type 1(CB1R),the synthesizing enzyme N-acyl phosphatidyl-ethanolamine phospholipase D(NAPE-PLD),and the degradation enzyme fatty acid amide hydrolase(FAAH)in the vervet monkey area V1 to better understand the mechanisms underlying the effects of eCB system modulation on cortical visual processing.Methods:Using Western blots and immunohistochemistry,we investigated the laminar and cellular expression patterns of CB1R,NAPE-PLD,and FAAH across the rostrocaudal axis of the vervet monkey(Chlorocebus sabaeus)primary visual cortex.Results:CB1R,NAPE-PLD,and FAAH were expressed in V1 throughout the rostrocaudal axis.CB1R showed very low staining in layer(L)4,with higher expression in all other layers,especially L1,followed by L2 and L3.NAPE-PLD and FAAH expression patterns were similar,but not quite as low in L4.CB1R,NAPE-PLD,and FAAH were localized in vGlut2-positive cells,representing glutamatergic projection neurons,and in somatostatin(SST)-positive cells,a class of GABAergic interneurons.Conclusions:The low level of CB1R in L4 indicates less direct endocannabinoid modulation of V1 afferents from the dLGN,but that greater modulation may occur via the higher expression of CB1R in L2 and L3 on the way to the dorsal and ventral visual streams.This is further supported by the higher expression of NAPE-PLD and FAAH in these layers.Expression in vGlut2-positive and SST-positive cells represents a role at both glutamatergic and GABAergic neurons.These data indicate that CB1R may influence the network of activity patterns in the visual streams after the visual information has reached V1,and thus may influence visual perception.

利莫那班对肝纤维化C57小鼠肝组织大麻素受体1及α-SMA表达的影响

目的研究大麻素受体1(CB1)拮抗剂利莫那班对肝纤维化模型C57小鼠肝组织中CB1、α-平滑肌肌动蛋白(α-SMA)表达的影响,及其抗肝纤维化的作用机制。方法 30只C57小鼠随机分为3组,分别为正常对照组、模型对照组及利莫那班组,每组10只。采用四氯化碳腹腔注射诱导形成小鼠肝纤维化模型。造模2周后于继续造模同时,正常对照组和模型对照组每天生理盐水灌胃,利莫那班组用利莫那班灌胃。第8周造模结束时处死小鼠,留取肝脏组织标本,分别进行HE和Masson三色染色,应用免疫组织化学方法检测肝组织中CB1和α-SMA的表达,并进行肝组织纤维化评分(S评分)。结果模型对照组和利莫那班组肝组织S评分、CB1和α-SMA阳性表达量均高于正常对照组(均P<0.05),利莫那班组均低于模型对照组(均P<0.05);正常对照组、模型对照组和利莫那班组CB1评分、α-SMA评分与S评分相互之间均呈正相关(均P<0.05)。结论肝组织CB1的激活可促进肝纤维化的形成,CB1拮抗剂利莫那班通过抑制CB1表达,进而抑制肝星状细胞的增殖和激活,从而起到抗肝纤维化的作用。

CB 1 cannabinoid receptor participates in the vascular hyporeactivity resulting from hemorrhagic shock in rats

Background Vascular hyporeactivity, which occurs in the terminal stage of hemorrhagic shock, is believed to be critical for treating hemorrhagic shock. The present study was designed to examine whether the CB1 cannabinoid receptor （CB1 R） was involved in the development of vascular hyporeactivity in rats suffering from hemorrhagic shock. Methods Sixteen animals were randomly divided into two groups （n=8 in each group）： sham-operated （Sham） and hemorrhagic shock （HS） groups. Hemorrhagic shock was induced by bleeding. The mean arterial pressure （MAP） was reduced to and stabilized at （25±5） mmHg for 2 hours. The vascular reactivity was determined by the response of MAP to norepinephrine （NE）. In later experiments another twelve animals were used in which the changes of CB1R mRNA and protein in aorta and superior mesenteric artery （SMA） were analyzed by RT-PCR and Western blotting. In addition, we investigated the effects of a CB1R antagonist on the vascular hyporeactivity and survival rates in rats with hemorrhagic shock. Survival rates were analyzed by the Fisher＇s exact probability test. The MAP response was analyzed by one-way analysis of variance （ANOVA）. Results Vascular hyporeactivity developed in all animals suffering from hemorrhagic shock. The expression of CBIR mRNA and protein in aorta and 2-3 branches of the SMA were significantly increased in the HS group after the development of vascular hyporeactivity when compared to those in Sham group. When SR141716A or AM251 was administered, the MAP response to NE was （41.75±4.08） mmHg or （44.78±1.80） mmHg respectively, which was higher than that in saline groups with （4.31±0.36） mmHg （P 〈0.01）. We also showed an increased 4-hour survival rate in the SR141716A or AM251-treated group with 20% or 30%, but with a statistically significant difference present between the AM251-treated and saline groups （P 〈0.05）. Conclusions CBIR is involved in vascular hyporeactivity resulting from hemorrhagic shock in rats, and CB1R antagonist may be useful in treating patients with traumatic, hemorrhagic shock who need field-rescue or initial treatment.

大麻素CB1受体对大鼠视网膜神经节细胞诱发动作电位的作用(英文)

激活大麻素CB1受体(CB1Rs)通过调控多种离子通道,从而调节脊椎动物视网膜的功能。本文旨在利用膜片钳全细胞记录技术,在大鼠视网膜薄片上研究CB1Rs对神经节细胞兴奋性的作用。结果显示,在电流钳制状态下,灌流CB1R激动剂WIN55212-2(WIN,5μmol/L)对神经节细胞的自发动作电位发放频率和静息膜电位均没有显著影响。在灌流液中加入CNQX,D-APV,bicuculline和strychnine以阻断神经节细胞的兴奋性和抑制性输入,灌流5μmol/L WIN对正向电流注入(+10pA到+100pA)诱发的动作电位的频率也没有显著影响。位相分析结果显示,触发动作电位的阈值电位和触发第一个动作电位的延迟时间在加入WIN前后也没有显著改变;然而,WIN显著降低动作电位的上升和下降相速率(±dV/dtmax),而且该作用可被CB1R拮抗剂SR141716所阻断。此外,在阻断突触输入的情况下,WIN对神经节细胞的膜电位也没有显著影响。以上结果提示,激活大麻素CB1Rs通过调控诱发动作电位,从而调节大鼠视网膜神经节细胞的兴奋性。

The endocannabinoid system:A new pharmacological target for obesity treatment?

Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and surgery, among which the diet is still the most common way. However, none of these therapeutic measures available is ideal, making it necessary to find an effective medical treatment. The endocannabinoid system, which is well known for its contributions in certain mental processes such as relaxation, amelioration of pain and anxiety, and sedation initiation, has been recently reported to play an essential role in regulating appetite and metabolism to maintain energy balance, leading to the belief that endocannabinoid system is closely related to obesity. This new discovery deepens our understanding of obesity, and provides us with a new direction for clinical obesity treatment. Rimonabant is an antagonist for CB1, and has entered the market in some countries. However, although effective as an anti-obesity drug, rimonabant also causes obviously adverse side-effects, thus is being doubted and denied for medical usage.

ACEA改善线粒体复合体活性诱导神经保护作用研究

目的:探讨线粒体复合体活性对大麻素CB1受体选择性激动剂ACEA神经保护作用的影响。方法:将原代大鼠皮层神经元分为4组:对照组(Control)、氧糖剥夺组(OGD)、ACEA+OGD组和溶剂(Vehicle)+OGD组,分别检测各组神经元损伤程度和线粒体复合体Ⅰ、Ⅱ和Ⅳ的活性。为进一步证实线粒体复合体活性对ACEA神经保护的影响,将原代大鼠皮层神经元分为5组:对照组(Control)、氧糖剥夺组(OGD)、ACEA+OGD组、线粒体复合体Ⅰ抑制剂(rotenone)+ACEA+OGD组和线粒体复合体Ⅱ抑制剂(TTFA)+ACEA+OGD组,检测和比较各组神经元细胞的损伤情况。结果:在OGD后24小时,ACEA明显增加神经元活性,减少LDH释放,降低神经元凋亡率(P<0.05),改善OGD损伤后线粒体复合体Ⅰ和Ⅳ的活性(P<0.05),而对复合体Ⅱ的活性没有影响;rotenone可以部分逆转ACEA的神经保护作用(P<0.05),但TTFA却没有这一作用。结论:ACEA可以诱导神经保护作用,其机制是与改善线粒体呼吸链复合体活性有关。