万拉法新对强迫游泳大鼠下丘脑和海马c-fos及c-jun蛋白表达的下调作用

目的　探索新一代抗抑郁药万拉法新对大鼠下丘脑和海马内ｃｆｏｓ 和ｃｊｕｎ 蛋白表达的影响。方法　采用特异性抗体的原位免疫细胞化学方法,在强迫游泳大鼠抑郁模型上,观察万拉法新慢性给药（ 腹腔内注射每日１ 次,连续７ 次）对大鼠游泳不动时间和下丘脑及海马核团ｃｆｏｓ 和ｃｊｕｎ 表达的影响;用图像分析技术对大鼠下丘脑室旁核（ Ｐ Ｖ Ｎ） 、视上核（ Ｓ Ｏ Ｎ） 和海马齿状回（ Ｄ Ｇ） 内的ｆｏｓ 和ｊｕｎ 阳性细胞的相对切面面积比和平均目标灰度进行分析。结果　强迫游泳可使大鼠下丘脑和海马内多个核团的ｃｆｏｓ 和ｃｊｕｎ 蛋白表达水平增加,而万拉法新明显缩短了强迫游泳大鼠的不动时间。图像分析结果提示,万拉法新使强迫游泳大鼠下丘脑 Ｐ Ｖ Ｎ 和 Ｓ Ｏ Ｎ 及海马 Ｄ Ｇ 内ｆｏｓ 和ｊｕｎ 阳性细胞相对切面面积比明显降低（ Ｐ＜００５） ,而平均目标灰度显著增加（ Ｐ＜ ００１） 。结论　下丘脑 Ｐ Ｖ Ｎ、 Ｓ Ｏ Ｎ 和海马 Ｄ Ｇ 可能是介导抗抑郁药抑制大鼠绝望行为的重要中枢核团,ｆｏｓ 和ｊｕｎ 蛋白可能是抗抑郁药发挥受体后作用的传导物质。     

鞘内注射曲马多对手术致痛大鼠脊髓c-fos蛋白表达的影响

目的在大鼠手术致痛模型中研究鞘内注射曲马多对脊髓c-fos蛋白的影响,探讨曲马多的超前镇痛效应。方法选取鞘内置管成功的32只雄性大鼠随机分为4组,每组8只,分别为假手术组(S组),实验对照组(EC组),术前曲马多组(TP20组,20μg/10μl)以及术后曲马多组(PT20组,20μg/10μl)。按照Brennan法制成大鼠切口痛模型,致痛2 h后,深麻醉大鼠,灌注固定,取脊髓L4～L6节段做c-fos免疫组化染色,观察大鼠脊髓c-fos蛋白表达的变化情况。结果 3个实验组的IOD值与假手术组相比较,均有不同程度的升高,差异有统计学意义(P0.05);TP20组、PT20组的IOD值与EC组相比较,均弱于实验对照组,差异有统计学意义(P0.05);TP20组的IOD值与PT20组相比较,差异有统计学意义(P0.05)。结论手术致痛前或后,鞘内注射曲马多均能抑制大鼠脊髓背角c-fos蛋白的表达;曲马多预先给药,抗伤害作用更好,具有超前镇痛效应。     

青藤碱对偏头痛模型大鼠脑干c-fos、c-jun表达的影响

目的 观察青藤碱对偏头痛模型大鼠脑于c-fos、c-jun表达的影响,了解青藤碱有无治疗偏头痛的作用.方法 60只健康Wistar大鼠随机分为空白对照组、模型组、舒马普坦对照组和青藤碱低、中、高剂量治疗组.皮下注射硝酸甘油(NTG)复制实验性偏头痛动物模型,药物干预4h后断头取脑.免疫组化法检测脑干c-fos、c-jun表达水平,显微镜下计数阳性细胞数.结果 与空白对照组相比较,模型组脑干c-fos、c-jun表达明显增多,差异有显著统计学意义(P＜0.01);与模型组相比较,青藤碱高、中、低剂量组和舒马普坦组脑干c-fos、c-jun表达明显减少,差异有显著统计学意义(P＜0.01);与舒马普坦组相比较,青藤碱高剂量组脑干c-fos、c-jun表达无差异,无统计学意义(P＞0.05).结论 青藤碱可能通过某些作用机制治疗偏头痛,阻断疼痛刺激信号传入脑干,抑制脑干c-fos、c-jun表达,减轻颅内疼痛长时程反应.     

帕金森病模型中热休克蛋白mRNA的表达

热休克蛋白（heat shock proteins, Hsp）在多种神经变性疾病模型中表达增加,并对细胞产生保护作用,因此我们用1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）和它的脑内活性代谢物1-甲基-4-苯基-吡啶离子（MPP＋）制备帕金森病（Parkinson disease, PD）动物和细胞模型,观察Hsp70和Hsp40的人类同源物HDJ-1 mRNA的动态变化和表达部位。     

养血清脑颗粒对硝酸甘油偏头痛模型大鼠三叉神经脊束核c-Fos蛋白表达的影响

目的观察养血清脑颗粒对硝酸甘油偏头痛模型大鼠脑干三叉神经脊束核c-Fos蛋白表达变化的影响。方法共90只雄性Wistar大鼠,分别于皮下注射硝酸甘油建立偏头痛动物模型之前(预防组)、模型制备后(治疗组)接受高剂量(0.32 g/ml)或低剂量(0.16 g/ml)养血清脑颗粒治疗,行为学评价后行免疫组织化学染色检测大鼠三叉神经脊束核神经元c-Fos蛋白表达水平。结果模型组大鼠各观察时间点(30、60、180 min)三叉神经脊束核神经元c-Fos蛋白表达水平与高、低剂量治疗组之间差异无统计学意义(均P0.05);但高于高剂量预防组(P=0.031,0.000,0.000),并于制模后60和180 min时高于低剂量预防组(均P=0.000);制模后30 min时高剂量预防组大鼠三叉神经脊束核神经元c-Fos蛋白表达水平低于低剂量预防组(P=0.029)。结论预防性应用养血清脑颗粒可以显著降低三叉神经脊束核c-Fos蛋白表达水平。提示养血清脑颗粒对偏头痛的预防作用大于治疗作用,其机制可能与降低三叉神经脊束核c-Fos蛋白表达水平有关。     

大鼠短暂脑缺血后海马区c-fos mRNA和FOS蛋白表达的时间规律

目的 探讨大鼠脑缺血-再灌注后脑海马区c-fos mRNA和FOS蛋白表达规律。方法 采用左侧大脑中动脉插入丝线结扎(LMCAO)方法制作大鼠脑缺血-再灌注缺血模型，分别按照30min至7d的不同灌注时间取材，应用原位杂交方法标记各实验组大鼠脑组织海马区c-for mRNA表达数量；应用免疫组化方法标记各实验组大鼠脑组织中海马区FOS阳性神经元数量。结果 c-fos mRNA阳性细胞在MCAO缺血再灌注1h开始表达，2h达高峰，至2d时c-fos mRNA阳性细胞表达基本消失；c-fos免疫活性细胞从2h开始表达，4h达高峰，4d时表达基本消失。脑缺血—再灌注后c-fos mRNA与FOS蛋白在海马区表达具有一定的时间上规律性。结论 本研究证明了大鼠MCAO缺血—再灌注后可诱发FOS蛋白和c-fos mRNA表达增加，且基因和蛋白的表达均具有一定的时间规律性和相关性。     

纳洛酮对大鼠脑缺血bcl-2和c-fos蛋白表达的影响

目的探讨纳洛酮对脑缺血再灌注的神经保护机制.方法采用线栓法制造的大鼠局灶性脑缺血再灌注模型,通过免疫组化法检测脑缺血不同再灌注时间以及经纳洛酮(5 mg/kg)预处理后bcl-2和c-fos蛋白的表达.结果脑缺血30 min再灌注6h,bcl-2和c-fos蛋白表达显著增加;再灌注24h,bcl-2蛋白持续增高,而c-fos蛋白基本降至正常.表明纳洛酮可显著上调脑缺血后bcl-2蛋白的表达,抑制c-fos蛋白的表达.结论纳洛酮的神经保护机制可能与其上调bcl-2蛋白,抑制c-fos蛋白表达有关.     

慢性应激抑郁模型大鼠强迫游泳后海马中c-fos的表达

目的 研究慢性应激对强迫游泳大鼠海马神经元c-fos表达的影响。方法 采用特异性抗体的免疫组织化学方法,在大鼠慢性应激抑郁模型基础上,观察急性强迫游泳应激后海马神经元c-fos的表达情况。结果 抑郁模型大鼠接受急性强迫游泳应激后,海马CA3区和齿状回(DG)内FOS蛋白的表达在各主要时点均比对照组降低,图像分析提示,上述两区域的FOS阳性神经元对应切面面积比明显降低(P<0.05),平均目标灰度值显著增加(P<0.05)。结论 慢性应激使大鼠在急性强迫游泳应激后海马CA3区和DG内c-fos的表达降低。     

类固醇肌病大鼠模型的建立及其骨骼肌病变的形态学研究

目的探讨不同剂量皮质类固醇激素以及合成代谢类激素对大鼠骨骼肌的影响。方法用不同剂量地塞米松(dexamethasone,DX),分别为5mg/kg腹腔注射,每日1次,连用14d和10、20、40、60、90mg/kg腹腔注射,每日1次,连用5d诱导类固醇肌病(steroidmyopathy,SM)的大鼠模型,观察其骨骼肌的病理改变;观察苯丙酸诺龙(NPP)对类固醇肌病的预防效果。结果DX5mg/kg腹腔注射,每日一次连续3天即足以造成大鼠的实验性SM模型,随剂量和疗程的增加,肌肉病变加重;病肌显示肌纤维萎缩和结构破坏,以Ⅱ型纤维为主,Ⅰ型纤维亦可受累,但程度较轻;Ⅰ型肌纤维在DX40、90mg/kg组中比例增高;NPP与DX5mg/kg同步使用未能防止SM的发生,但减轻了肌纤维的坏变程度。结论大鼠实验性SM模型的病肌以肌纤维萎缩为主,伴纤维结构破坏,病变随激素剂量和疗程的递增而加剧;Ⅰ型和Ⅱ型肌纤维均可受累;合成代谢类激素不能防止SM的发生。     

神经调节蛋白/表皮生长因子受体通路在偏头痛大鼠模型中的作用

目的 探讨神经调节蛋白(neuregulin,NRG)/表皮生长因子受体(epidermal growth factor re-ceptor,ErbB)通路在偏头痛大鼠模型中的作用.方法 将Wistar大鼠随机分为对照组、模型组、空载(空慢病毒载体)组、NRG沉默(含NRG基因干扰片段的慢病毒载体)组、NRG1(0.0...     

Regulation of c-fos gene expression by lipopolysaccharide and cycloheximide in C6 rat glioma cells

The effect of lipopolysaccharide (LPS) on the expression of immediate early genes, such as c-fos and c-jun, was examined in C6 rat glioma cells. LPS (1 microg/ml) alone did not affect c-fos mRNA level. LPS, however, transiently increased c-jun mRNA level. Cycloheximide (CHX, 20 microM), a protein synthesis inhibitor, alone caused increases of c-fos and c-jun mRNA levels. LPS showed a potentiating effect in the regulation of c-fos mRNA level, whereas LPS showed an additive action for the regulation of CHX-induced c-jun mRNA expression. To determine if CREB and mitogen-activated protein kinases (MAPKs) are involved in the regulation of c-fos mRNA expression by LPS and CHX, Western blot was carried out using the phosphorylated form of antibodies against ERK, JNK, p38, and CREB. LPS transiently increased the phosphorylation of p38-MAPK and CREB. In addition, LPS alone elevated phosphorylation of ERK (p44/p42) MAPK in a time-dependent manner. Furthermore, LPS plus CHX enhanced phosphorylation of ERK, p38, and CREB in a synergistic manner. Our results suggest that the phosphorylation of ERK, p38, and CREB may be involved in the regulation of synergistic c-fos mRNA expression induced by LPS plus CHX in C6 rat glioma cells.     

Post-ischemic and kainic acid-induced c-fos protein expression in the rat hippocampus

The c-fos protein is a gene regulatory third messenger involved in long-term responses of cells to various stimuli. It can be used as a marker of neuronal activity. In the present immunohistochemical study the presence of c-fos protein (FP) in the rat brain from 1 h to 14 days after 10 min of cerebral ischemia was compared with that 3 h after an intraventricular injection of kainic acid. The kainic acid injection resulted in staining of dentate hilar cells, granule cells and hippocampal interneurones. The postischemic changes at Day 1 were sporadic CA1 pyramidal cells expressing the FP. At Day 2 FP was expressed with variable intensity in many pyramidal cells in the CA1. At Day 3 many necrotic CA1 pyramidal cells were seen. They did not express the FP, and the expression was less intense and found in fewer cells than at Day 2. At Days 3, 7 and 14 there was increasing gliosis without c-fos expression in the CA1. The study demonstrates a delayed postischemic synthesis of the gene regulatory protein c-fos preceding the necrosis in the selectively vulnerable CA1 region.     

Regulation of expression of cytochrome P-450 2D mRNA in rat brain with steroid hormones

Cytochrome P-450 2D is a subfamily of the cytochrome P-450-dependent mixed function oxidase system which is widely distributed in the various tissues of mammals. Sex steroid hormones have been shown to affect the expression of CYP2D in rat brain. Testosterone treatment of ovariectomized female rats elicits a dramatic increase in CYP2D expression, estrogen treatment brings about a modest increase in brain CYP2D expression and reduces the increase in CYP2D expression elicited with testosterone when the two hormones are coadministered. Polymerase chain reaction (PCR) has been used in our laboratory, as well as other laboratories, to measure the low levels of message for various P-450s in brain [Hodgson, A.V., White, T.B., White, J.W., Strobel, H.W., 1993. Expression analysis of the mixed function oxidase system in rat brain by the polymerase chain reaction. Mol. Cell. Biochem. 120, 171–179; Omiecinski, C.J., Redlich, C.A., Costa, P., 1990. Induction and developmental expression of cytochrome P450IA1 messenger RNA in rat and human tissues: detection by the polymerase chain reaction. Cancer Res. 50, 4315–4321]. In this study, competitive PCR (cPCR) approaches have been used to determine effects of progesterone and testosterone on CYP2D expression levels in brains of intact and ovariectomized female rats. When administered for seven treatments, testosterone significantly increases the expression of CYP2D in brain from intact female rats, while repeated treatment with progesterone elicits the opposite effect. Coadministration of testosterone and progesterone causes an intermediate effect such that the net result is an increase in expression only slightly above control levels. Interestingly, when ovariectomized female rats treated with testosterone and progesterone are used as a source of brain tissue for RNA preparation a similar trend toward an intermediate value is seen but the net result is an expression level of CYP2D below the control value. This approach utilizes cPCR to analyze the levels of CYP2D mRNA, semi-quantitatively and quantitatively, in the brains of female intact and ovariectomized Sprague–Dawley rats treated with testosterone, progesterone, a combination of the two or corn oil.     

顺行性灌注二磷酸果糖对深低温停循环大鼠脑组织c-fos mRNA表达的影响

目的研究大鼠深低温停循环后脑组织中c-fos mRNA表达的时效关系。方法建立大鼠深低温停循环模型，用RT-PCR方法检测不同时间点大鼠海马组织内c-fos mRNA表达的强弱。结果c-fos mRNA的表达增强，在复温后即刻达高峰，与对照组相比，差异无统计学意义；在复温1h后开始减弱，6h最低，但均强于对照组，P〈0.05。结论（1）深低温停循环引起c—fos mRNA的表达；（2）含有1．6二磷酸果糖的冷脑保护液间断灌流，可以增强c—fos基因的表达。     

脑缺血再灌注后脑组织c—fos基因表达与东菱克栓酶的影响

本实验采用线栓法制成大鼠大脑中动脉缺血再灌注模型,用地高辛精标记的c—fos探针进行原位杂交。结果示缺血再灌注组栓塞侧皮层及海马c—fos基因表达显著增多,图像分析灰阶值为118.6±5.1,对侧为159.6±3.1(P<0.001),东菱克栓酶组栓塞侧皮层及海马c—fos基因表达亦增多,灰阶为131.0±4.5,对侧为164.6±4.0(P<0.001)。克栓酶组与缺血再灌注组比较,栓塞侧东菱克栓酶组c—fos基因表达显示低于缺血再灌组(P<0.05),而两组栓塞对侧比较无显著差异。本实验表明,脑缺血再灌注后脑组织的c—fos基因表达显著增多,东菱克栓酶能抑制缺血后c—fos基因的表达,这可能是其治疗缺血性脑血管病的机理之一。     

局灶脑缺血后胰岛素对c-fos、c-jun基因表达的影响

目的　观察胰岛素对局灶缺血性脑组织 c- fos、c- jun基因表达的影响。方法　 30只肾血管性高血压大鼠随机分 4组 ,在大脑中动脉闭塞 (MCAO)后即予胰岛素 (2 .1IU / kg,A组 )、葡萄糖和胰岛素 (2 .1IU / kg,B组 ;4.5 IU / kg,C组 )、生理盐水 (D组 ) ,采用原位杂交技术检测各组 MCAO后 3h脑组织 c- fos、c- jun基因的表达。结果　与其它组比较 ,A组血糖下降明显 (P<0 .0 1) ,MCAO后 1h、2 h、3h分别为 3.80± 0 .3mm ol/ L、3.6 8± 0 .3mm ol/L、3.6 9± 0 .3mm ol/ L。 c- fos m RNA被诱导出现于缺血侧皮层和尾壳核 ,C组 c- fos m RNA明显增加 (P<0 .0 1) ,皮层和尾壳核的灰度为 12 3.5 8± 8.72、141.0 8± 8.49;c- jun m RNA仅出现于缺血侧皮层 ,A、B、C组的 c- jun m RNA均增加明显 ,灰度分别为 171.82± 7.33、172 .5 6± 7.91、15 5 .0 9± 7.91,其中 C组的增加更为显著 (P<0 .0 1)。结论　胰岛素促进缺血脑组织 c- fos、c- jun基因表达可能是其神经保护作用机制之一。     

Amphetamine-induced c-fos mRNA expression is altered in rats with neonatal ventral hippocampal damage

To further characterize the mechanisms underlying enhanced dopamine-related behaviors expressed during adulthood in rats with neonatal excitotoxic ventral hippocampal (VH) damage, we studied the expression of c-fos mRNA in these rats after a single saline or amphetamine (AMPH) (10 mg/kg, i.p.) injection using in situ hybridization. The VH of rat pups was lesioned with ibotenic acid on postnatal day 7 (PD7). At the age of 90 days, rats were challenged with AMPH or saline, and the expression of c-fos mRNA using an oligonucleotide probe was assessed 30, 90, and 180 min later. AMPH significantly increased c-fos mRNA expression in medial prefrontal cortex, piriform cortex, cingulate cortex, septal region, and dorsolateral and ventromedial striatum in control and lesioned rats. However, this response to AMPH was attenuated 30 min after AMPH injection in all of these regions in the lesioned as compared to the sham-operated rats. No significant changes were seen at other time points. These results indicate that the neonatal VH lesion alters time-dependent intracellular signal transduction mechanisms measured by AMPH-induced c-fos mRNA expression in cortical and subcortical brain regions. Changes in c-fos mRNA expression in this putative animal model of schizophrenia may have implications for long-term alterations in cellular pheno type because of altered regulation of certain target genes. (This article is a US Government work and, as such, is in the public domain in the United States of America.) © 1996 Wiley-Liss, Inc.