Effect of tetrahydropalmatine analogs on Fos expression induced by formalin pain 1

目的：研究四氢巴马汀（ＴＨＰ）同类物对福尔马林致痛诱导的Ｆｏｓ蛋白表达的影响,以阐明ＴＨＰ同类物的镇痛机制．方法：在右后肢脚掌皮下注射５％福尔马林５０μＬ,诱发炎性疼痛,用免疫组织化学方法观察Ｆｏｓ蛋白表达．结果：腹腔注射ＴＨＰ同类物和Ｄ２受体拮抗剂螺哌隆诱导的Ｆｏｓ蛋白表达主要位于纹状体和伏膈核．Ｄ２受体激动剂喹吡罗可阻滞ｌＴＨＰ和螺哌隆诱导的Ｆｏｓ蛋白表达．ＴＨＰ同类物明显增加脑干下行痛觉调制系统的Ｆｏｓ蛋白表达,并能明显抑制福尔马林诱导的脊髓背角浅层和深层的Ｆｏｓ蛋白表达．结论：ＴＨＰ同类物通过阻滞纹状体和伏膈核的Ｄ２受体,加强脑干下行痛觉调制系统的功能,抑制外周痛觉信息在脊髓水平的传入,达到它们的镇痛作用．     

NMDA receptors mediating Fos expression in rat spinal cord induced by subcutaneous injection of formalin

目的：研究ＮＭＤＡ和非ＮＭＤＡ受体在疼痛刺激诱发脊髓Ｆｏｓ表达中的作用．方法：大鼠单侧后足跖部皮下注射２％福尔马林，免疫组化显示Ｆｏｓ的表达．结果：注射福尔马林２ｈ后，Ｆｏｓ阳性神经元集中分布在同侧脊髓背角Ⅰ层的内侧和Ⅱ层的浅部．脊髓鞘内给予ＮＭＤＡ受体拮抗剂ＡＰＶ（１０μＬ，００１，０１，１ｇ·Ｌ－１）剂量依赖性引起福尔马林诱发的背角Ｆｏｓ阳性细胞数量减少（Ｐ＜００１）；非ＮＭＤＡ受体拮抗剂ＤＮＱＸ对Ｆｏｓ表达无明显影响．结论：ＮＭＤＡ受体介导福尔马林致痛诱导的脊髓Ｆｏｓ表达．     

脊髓以上多巴胺D2受体介导左旋四氢巴马汀的镇痛作用

目的：研究ＤＡ受体与左旋四氢巴马汀（ｌ－ＲＨＰ）镇痛作用的关系,以阐明ｌＴＨＰ的镇痛机制。方法：腹腔（ＩＰ）与鞘内（ＩＴＨ）给药,以大鼠甩尾反应观测热伤害性致痛阈。结果：ｉｐ ｌ－ＴＨＰ或Ｄ２受体拮抗剂螺哌隆产生剂量依赖性镇痛效应,并能被Ｄ２受体激动剂喹吡罗翻转,蛤不被纳洛酮翻转。而ｉｔｈ ｌＴＨＰ或螺哌隆无镇痛效应,但它们能拮抗ｉｔｈ喹吡罗引起的镇痛效应。结论：激动脊髓Ｄ２受体或阻滞脊髓以上水平     

Effects of tetrahydroprotoberberines on dopamine D_2 receptors in ventral tegmental area of rat

阐明四氢原小檗碱同类物（ＴＨＰＢ）对大鼠中脑腹侧被盖区（ＶＴＡ）多巴胺（ＤＡ）受体的作用特性，并比较它们的作用强度．方法：采用大鼠在体胞外单位放电记录．结果：观察了１１个ＴＨＰＢ均可完全地翻转ＤＡ受体激动剂阿扑吗啡（２０μｇ·ｋｇ－１）所产生的放电抑制作用，为Ｄ２受体拮抗剂的作用特性．ＴＨＰＢ对Ｄ２受体的作用与Ｃ２位上的ＯＨ基团有密切的关系．它们的作用强度（ＥＤ５０，μｇ·ｋｇ－１）：ＴＨＰＢ１４３（５６）＞ＳＰＤ（８５）＞Ｉｓｏ（１７０）＞ＴＨＰ（３３）＞ＴＨＢ（４８）＞ＴＨＰＢ１８（６６）＞ＴＨＰＢ１（１７９）＞ＴＨＰＢ１９（４０８）＞ＴＨＰＢ１２６（５１０）＞ＴＨＰＢ１０４（１０１９）＞ＴＨＰＢ１０（４８１５）．结论：１１个ＴＨＰＢ均为ＶＴＡＤ２受体拮抗剂，以Ｃ２位上有ＯＨ基团的ＴＨＰＢ１４３作用最强．     

四氢原小檗碱同类物与多巴胺D2受体作用的定量构效关系

四氢原小檗碱同类物是一类新型的脑内多巴胺受体阻滞剂。本文用Ｆｒｅｅ－Ｗｉｌｓｏｎ法研究了一组四氢原小檗碱同类物与多巴胺Ｄ２受体作用的定量构效关系。结果表明：Ｃ－２位上的ＯＨ和Ｃ－１２位上的Ｃｌ使四氢原小檗碱同类物与多巴胺Ｄ２受本的亲和力增强,Ｃ－１１位上的立体位阻可能使亲和力降低,Ｃ－１０位上的取代基可能对亲和力影响不大。     

大鼠伏膈核内多巴胺受体与电针镇痛的关系

目的：研究多巴胺受体拮抗剂左旋四氢巴马汀（ｌ－ＴＨＰ）加强电针镇痛（ＥＡＡ）的原理，阐明中枢神经系统内多巴胺（ＤＡ）系统在ＥＡＡ中的作用，方法：分别将Ｄ１受激动剂ＳＫ＆Ｆ－３８３９３和Ｄ２受体激动剂ｑｕｉｎｐｉｒｏｌｅｈｙｄｒｏｃｈｌｏｒｉｄｅ（Ｑｕｉ）注射入大鼠伏膈核，观察对ＥＡＡ及ｌ－ＴＨＰ加强ＥＡＡ的作用。结果：ＳＫ＆Ｆ－３８３９３（５μｇ，１０μｇ）明显对抗ｌ－ＴＨＰ加强ＥＡＡ的作用，１０     

人TNFβ缺失体融鸽主其产物一步纯化法研究

本文交ｈＴＮＦβＮ端缺失２３ａａ的缺失体基因克隆于ｐＥＴ－２８－Ｃ＾（＋）表达戴本,构建成Ｔ７ｌａｃ启动子控制下Ｈｉｓ６－ＴＮＦβ融合表达质粒,转化到Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）,经ＩＰＴＧ诱导表达,Ｈｉｓ６－ＴＮＦβ表达量约占总菌体蛋白的２５％,Ｍｒ２０５００。表达产物大部分以包涵体形式存在。包涵体经过洗涤,７０ｍｏｌ／Ｌ脲变性溶解,用Ｍｉ＾２＋亲和层析一步快速纯化,所得Ｈｉｓ６－ＴＮＦβ的纯     

FADD研究进展

ＦＡＤＤ又名Ｍｏｒｔ１,它是一种死亡域蛋白 ,与Ｆａｓ胞浆区呈特异性结合 ,故名Ｆａｓ相关死亡域蛋白 (Ｆａｓ -ａｓｓｏｃｉａｔｅｄｗｉｔｈｄｅａｔｈｄｏｍａｉｎｐｒｏｔｅｉｎ)［１］:ＦＡＤＤ是一种胞浆蛋白 ,死亡受体配体或激动抗体与细胞表面死亡受体结合后 ,募集ＦＡＤＤ ,构成信号复合体 ,转导凋亡信号 ,因此它在凋亡信息转导过程中起着中断作用 ,故认为它是一种适配子蛋白或连续蛋白 (Ａｄａｐｔｅｒｐｒｏｔｅｉｎ) ［２］。Ｃｈｉｎｎａｉｙａｎ、Ｂｏｌｄｉｎ等［１、３］于１９９５年几乎同时筛选出一个与Ｆａｓ胞浆区能…     

抗抑郁剂慢性给药对强迫游泳大鼠下丘脑c-Fos蛋白表达水平的下调作用

应用ｃ－Ｆｏｓ蛋白免疫组织化学定位观察的方法，在强迫游泳大鼠抑郁模型上，观察地昔帕明（５，２０ｍｇ·ｋｇ－１），吗氯贝胺（１０，４０ｍｇ·ｋｇ－１）和氟西汀（５，２０ｍｇ·ｋｇ－１）慢性给药（ｉｐ每日１次，连续７ｄ）对大鼠游泳不动时间和下丘脑核团ｃ－Ｆｏｓ蛋白表达水平的影响．结果表明：强迫游泳可使大鼠下丘脑多个核团的ｃ－Ｆｏｓ蛋白表达水平明显升高，而地昔帕明，吗氯贝胺，氟西汀明显缩短强迫游泳大鼠的不动时间，并选择性地使强迫游泳诱导增加的下丘脑室旁核Ｆｏｓ样免疫阳性神经元数目明显减少．提示下丘脑室旁核可能是介导抗抑郁剂抑制大鼠绝望行为的中枢部位之一．Ｆｏｓ蛋白可能是不同类型抗抑郁剂共同的受体后信号转导物质     

槲皮素对PDGF诱导的NIH 3T3细胞增殖的影响

目的研究酪氨酸蛋白激酶抑制剂槲皮素对ＰＤＧＦ诱导的ＮＩＨ３Ｔ３细胞增殖的影响。方法采用ＭＴＴ比色法、Ｇｉｅｍｓａ染色、流式细胞术（测定ＤＮＡ含量的变化及增殖细胞核抗原的表达）、及ＷｅｓｔｅｒｎＢｌｏｔ分析技术对ＰＤＧＦ诱导的ＮＩＨ３Ｔ３细胞增殖进行分析。结果与对照组相比，槲皮素处理可显著地抑制ＰＤＧＦ诱导的ＮＩＨ３Ｔ３细胞增殖，而且主要是细胞周期Ｓ期抑制，ＷｅｓｅｔｅｒｎＢｌｏｔ分析提示槲皮素可明显地抑制ＰＤＧＦ诱导的ＮＩＨ３Ｔ３细胞酪氨酸磷酸化程度。结论酪氨酸蛋白激酶抑制剂可显著地抑制ＰＤＧＦ诱导的ＮＩＨ３Ｔ３细胞增殖，可能是通过抑制酪氨酸蛋白激酶活性来完成。     

Involvement of Adenosine A2A Receptors in the Induction of C-Fos Expression by Clozapine and Haloperidol

Acute administration of the atypical antipsychotic clozapine induced a regional pattern of c-fos expression characterized by an increase in Fos–like-immunoreactivity (FLI) in the prefrontal and prelimbic/infralimbic cortices, nucleus accumbens, and lateral septum and a weak activation of FLI in the striatum. Haloperidol, similarly to clozapine, increased FLI in the nucleus accumbens and lateral septum, but it did not induce FLI in prefrontal and prelimbic/infralimbic cortices. Moreover, haloperidol increased FLI in the striatum. To gain insight into the mechanism by which clozapine and haloperidol induced FLI in these brain structures, we evaluated whether blockade of adenosine A_2A receptors could influence these effects. The selective and high-affinity A_2A receptor antagonist SCH 58261 (5 mg/kg) completely abolished FLI induced by clozapine (20 mg/kg) in all subdivisions of the nucleus accumbens (rostral pole, shell and core) and striatum, but did not affect the number of Fos-like positive neurons in the prefrontal, prelimbic/infralimbic cortices, and lateral septum. SCH 58261 (5 mg/kg) reduced FLI induced by haloperidol (0.1 mg/kg) in the striatum, lateral septum, and all nucleus accumbens subdivisions. In contrast, FLI induced by 0.5 mg/kg of haloperidol in the shell and core of the nucleus accumbens was not affected by SCH 58261. The results show that adenosine A_2A receptors participate in the induction of FLI by clozapine and haloperidol and support the concept that A_2A receptors are involved in the mediation of antipsychotic effects.     

Effects of perospirone, a novel 5-HT2 and D2 receptor antagonist, on Fos protein expression in the rat forebrain.

The effects of perospirone, a novel 5-HT2 and D2 receptor antagonist, on Fos protein expression in the nucleus accumbens (NA) and dorsolateral striatum (DLSt) were compared with those of typical (i.e., haloperidol and fluphenazine) and atypical (i.e., clozapine and risperidone) antipsychotics using immunohistochemical techniques in rats. Perospirone and other antipsychotics tested at doses that exerted D2 blocking actions increased Fos-like immunoreactivity both in the NA and DLSt. However, the levels of Fos expression in the DLSt induced by perospirone and clozapine were less than those induced by haloperidol and fluphenazine. When compared the differences in numbers of Fos-positive neurons between in the NA and DLSt, perospirone, clozapine, and risperidone preferentially increased Fos expression in the NA. These findings suggest that perospirone has a preferential action on the mesolimbic (vs. nigrostriatal) dopaminergic system in inducing Fos protein in the rat brain, which may be related to its atypical antipsychotic properties.     

No change in neostriatal D-2 dopamine receptors after NMDA lesions of rat prefrontal cortex

D-2 dopamine receptor function in rat nucleus accumbens and anterior corpus striatum was examined 6-8 days following N-methyl-D-aspartate lesions confined to medial prefrontal cortex. Lesions failed to alter the affinity or density of D-2 receptors labelled by [3H]spiperone in membrane preparations of both subcortical areas. Locomotor activity and stereotyped behaviours induced by the D-2 agonist, LY-171555, were also not significantly altered in lesioned animals. These results suggest that D-2 dopamine receptors of nucleus accumbens and anterior corpus striatum are not localized on corticofugal afferents from medial prefrontal cortex.     

鞘内注射大麻酚类抑制大鼠脊髓内有毒刺激引起的Fos蛋白样免疫反应及其与吗

AIM: To determine whether cannabinoids suppress noxious stimulus-evoked Fos protein-like immunoreactivity (FLI) through direct actions at the spinal level. METHODS: Rats were implanted with intrathecal (ith) catheters at least one week prior to evaluation in the formalin test. Effects of the cannabinoid agonist, CP55,940 (80 micrograms ith) on formalin pain and FLI in rat spinal cord were compared with that of the prototypic narcotic analgesic, morphine (20 micrograms ith). CP55,940 suppressed pain behavior and FLI induced by intraplantar formalin. The cannabinoid suppressed Fos in the neck region of the dorsal horn and in the ventral horn, but not in the nucleus proprius. The efficacy of the cannabinoid in suppressing FLI in these laminae and pain behavior was comparable to morphine administered via the same route. However, only morphine suppressed FLI in the superficial dorsal horn relative to vehicle treatment. CONCLUSION: Cannabinoids suppress nociceptive processing, in part, through actions at the spinal level. However, morphine showed greater potency and efficacy than CP55,940 in suppressing formalin-induced FLI following spinal administration.     

Effects of l-stepholidine on forebrain Fos expression: comparison with clozapine and haloperidol.

l-Stepholidine (SPD) is a tetrahydroprotoberberine alkaloid and a mixed dopamine D1 agonist/D2 antagonist. Preliminary clinical trials suggest that SPD improves both positive and negative symptoms of schizophrenia without producing significant extrapyramidal side effects. Here, we report that SPD mimics the effect of the atypical antipsychotic drug clozapine, preferentially increasing Fos expression in corticolimbic areas. Thus, at 10 mg/kg (i.p.), SPD induced Fos expression in the medial prefrontal cortex (mPFC), nucleus accumbens (NAc), and lateral septal nucleus (LSN) without significantly affecting the dorsolateral striatum (DLSt). At higher doses (20-40 mg/kg), SPD also increased Fos expression in the DLSt. The increase, however, was less pronounced than the increase seen in the NAc. Within the NAc, SPD also induced more Fos expression in the shell than in the core. In all subcortical areas examined, the Fos expression induced by SPD was mimicked by the D2 antagonist sulpiride and reversed by the D2 agonist quinpirole, suggesting that the effect is due to blockade of D2-like receptors by SPD. In the mPFC, however, the effect was not mimicked by sulpride or reversed by quinpirole. It was also not mimicked by the D1 agonist SKF38393 or SKF38393 plus sulpride, and not reversed by the D1 antagonist SCH23390. These results suggest that, in the mPFC, SPD may induce Fos expression through a non-DA mechanism. Whether the mechanism involves an interaction of SPD with other neurotransmitters such as 5-HT and norepinephrine remains to be determined.     

Pseudoephedrine, a sympathomimetic agent, induces Fos-like immunoreactivity in rat nucleus accumbens and striatum.

The pharmacological properties of the ephedrine derivative pseudoephedrine were investigated at the nuclear level. Following intraperitoneal injection of Sprague Dawley rats with pseudoephedrine, Fos induction was measured in various brain areas by Western blots and immunocytochemistry. Pseudoephedrine induced Fos-like immunoreactivity in the nucleus accumbens and striatum in a time and concentration-dependent manner with maximal effect at 60 mg/kg 2 h after injection. Immunocytochemical studies confirmed that the majority of the signal was detectable in the nucleus accumbens and striatum. Pre-injection with the D1 dopamine receptor antagonist SCH23390 partially and completely blocked pseudoephedrine-induced Fos-like immunoreactivity in the striatum and nucleus accumbens, respectively, suggesting that the action of pseudoephedrine is mediated via dopamine release and results in the activation of D1 dopamine receptors. With the exception of the higher doses required, the actions of pseudoephedrine were similar to those previously described for the psychostimulant amphetamine.     

Effects of chronic treatment of MPTP monkeys with bromocriptine alone or in combination with SKF 38393

Eight monkeys developed a severe parkinsonian syndrome after i.v. administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Treatment with bromocriptine (5 mg/kg) relieved the parkinsonian symptoms, but the efficacy of this treatment appeared to decrease slightly with time. The addition of SKF38393 (5 mg/kg) to the bromocriptine treatment in four monkeys prevented and even reversed the tendency to decreased efficacy with an increased response in three out of four animals. Neither of these two treatments induced dyskinesia in these monkeys. Dopamine D1 and D2 receptors, assayed in the caudate nucleus, the putamen and nucleus accumbens with [3H]SCH 23390 and [3H]spiperone binding respectively, were not significantly different in MPTP monkeys treated with bromocriptine or with bromocriptine plus SKF38393. Monkeys in the two treatment groups had a similar extent of denervation of the striatum and accumbens as assessed by the content of dopamine and its metabolites. These results suggest that in MPTP monkeys, the behavioral response to the D2 agonist bromocriptine can be enhanced by concomitant activation of the D1 receptors while this combination of agonists does not induce dyskinesia. The addition of the D1 agonist does not appear to cause further alteration of the D1 or D2 receptors.     

Adenosinergic modulation of 3α-hydroxy-5α-pregnan-20-one induced catalepsy in mice

  Rationale: Neurosteroid 3α, 5α THP, a positive allosteric modulator of the GABA_A receptor Cl^– ionophore complex, induces catalepsy-like dopamine antagonists, adenosine agonists or GABA agonists. Adenosine and dopamine receptors are co-localized on GABAergic neurons in the striatum and regulate GABA-mediated neurotransmission. Moreover, the antagonistic interactions between specific subtypes of adenosine and dopamine receptors are involved in motor depressant or motor stimulant effects of adenosine receptor agonists or antagonists, respectively. Such interaction may modulate neurosteroid-induced catalepsy. Objective: This study examined the modulation of 3α, 5α THP-induced catalepsy by adenosinergic agents. Methods: Catalepsy induced by 3α, 5α THP (2–8 μg, ICV) was assessed by bar test periodically up to 3 h in mice. Adenosine A₁, A_2A or A₃ receptor agonists or antagonists were given IP or ICV prior to 3α, 5α THP. Some animals received IP dopamine D₂ receptor agonist or antagonist 30 min prior to above combination treatment. Results: Adenosine A₁, A_2A, and A₃ receptor agonists potentiated, whereas adenosine A_2A receptor antagonists, but not A₁ antagonists, reversed 3α, 5α THP-induced catalepsy. These effects of adenosine agonists and antagonists were abolished by prior administration of bromocriptine, the dopamine D₂ receptor agonist and spiperone, the dopamine D₂ receptor antagonist, respectively. Conclusions: These findings suggest specific adenosine-dopamine receptor interaction in the striatum to modulate 3α, 5α THP-induced catalepsy. Received: 1 November 1998 / Final version: 5 January 1999     

Armodafinil promotes wakefulness and activates Fos in rat brain

Modafinil increases waking and labeling of Fos, a marker of neuronal activation. In the present study, armodafinil, the R-enantiomer of racemic modafinil, was administered to rats at 30 or 100 mg/kg i.p. about 5 h after lights on (circadian time 5 and near the midpoint of the sleep phase of the sleep:wake cycle) to assess its effects on sleep/wake activity and Fos activation. Armodafinil at 100 mg/kg increased wakefulness for 2 h, while 30 mg/kg armodafinil only briefly increased wakefulness. Armodafinil (30 and 100 mg/kg) also increased latencies to the onset of sleep and motor activity. Armodafinil had differential effects in increasing neuronal Fos immunolabeling 2 h after administration. Armodafinil at 100 mg/kg increased numbers of Fos-labeled neurons in striatum and anterior cingulate cortex, without affecting nucleus accumbens. Armodafinil at 30 mg/kg only increased numbers of light Fos-labeled neurons in the anterior cingulate cortex. In brainstem arousal centers, 100 mg/kg armodafinil increased numbers of Fos-labeled neurons in the tuberomammillary nucleus, pedunculopontine tegmentum, laterodorsal tegmentum, locus coeruleus, and dorsal raphe nucleus. Fos activation of these brainstem arousal centers, as well as of the cortex and striatum, is consistent with the observed arousal effects of armodafinil.     

Definition of the in-vivo accumulation of [3H]spiperone in brain using haloperidol and sulpiride to determine functional dopamine receptor occupation

The in-vivo administration of [3H]spiperone caused an accumulation of radioactivity in the substantia nigra, tuberculum olfactorium, nucleus accumbens, striatum and frontal cortex when compared with cerebellar levels. Haloperidol (0.01-1.0 mg kg-1 i.p.) dose-dependently prevented the accumulation of [3H]spiperone in the substantia nigra, tuberculum olfactorium, striatum and nucleus accumbens. Sulpiride (10-160 mg kg-1 i.p.) dose-dependently prevented the accumulation of [3H]spiperone only in the substantia nigra. The effects of sulpiride on other areas were not consistent; there was a suggestion of a reduction in the accumulation of [3H]spiperone in tuberculum olfactorium and striatum, but not in nucleus accumbens. Neither haloperidol (0.01-1.0 mg kg-1 i.p.) nor sulpiride (10-160 mg kg-1 i.p.) caused displacement of [3H]spiperone from the frontal cortex. Both haloperidol (0.01-0.5 mg kg-1) and sulpiride (10-80 mg kg-1) increased striatal and mesolimbic HVA concentrations. Haloperidol potently blocked apomorphine-induced stereotypy but sulpiride was only effective at the highest dose employed. The functional effect produced by haloperidol correlated with its ability to define [3H]spiperone binding in-vivo to dopamine receptors in the substantia nigra, striatum and tuberculum olfactorium. In contrast, there was no correlation between functional effect of sulpiride and its ability to define [3H]spiperone binding in-vivo.