Effects of acute and chronic administration of MK-801 on c-Fos protein expression in mice brain regions implicated in schizophrenia and antagonistic action of clozapine

Objective To investigate the effects of acute and chronic administration of the non-competitive NMDA receptor antagonists MK-801 on c-Fos protein expression in different brain regions of mice and an-tagonistic action of clozapine.Methods Immunohistochemistry was used to detect the expression of c-Fos protein.Results MK-801(0.6 mg·kg-1)acute administration produced a significant increase in the expression of c-Fos protein in the layers Ⅲ-Ⅳ of posterior cingulate and retrosplenial(PC/RS)cortex,which was consistent with the previous reports.Moreover,we presented a new finding that MK-801(0.6 mg·kg-1)chronic administration for 8 days produced a significant increase of c-Fos protein expression in the PC/RS cortex,prefrontal cortex(PFC)and hypothalamus of mice.Among that,c-Fos protein expression in the PC/RS cortex of mice was most significant.Compared acute administration with chronic administration,we found that MK-801 chronic administration significantly increased the expression of c-Fos protein in the PC/RS cortex,PFC and hypothalamus.Furthermore,pretreatment of mice with clozapine significantly decreased the expression of c-Fos protein induced by MK-801 acute and chronic administration.Conclusions Marked expression of c-Fos protein induced by MK-801 is associated with neurotransmitters' change noted in our previous studies,and c-Fos protein,the marker of neuronal activation,might play an important role in the chronic pathophysiological process of schizophrenic model induced by NMDA receptor antagonist.

EXPRESSION AND SIGNIFICANCE OF ONCOPROTEIN p16 AND FOS IN OSTEOSARCOMA

Objective: To investigate p16, C-fos protein expression and their relationships in osteosarcoma. Methods: Immunohistochemical technique (SABC) was used to detect p16 and C-fos protein expression in 41 cases of osteosarcoma. Results: The positive rates of p16 and C-fos protein expression were 51.2% and 82.9% respectively. Their expression was not correlated to pathological subtype, but correlated to clinic grade, and the latter was associated with tumor metastasis. There was a negative correlation between p16 and C-fos protein expression. Conclusion: The alteration of p16 and C-fos protein expression may be related to the tumorigenesis and development of osteosarcoma, and C-fos proteins may take part in osteosarcoma metastasis. These data will offer useful helpness to determine the prognosis of osteosarcoma.

MicroRNA-185-5p mediates regulation of SREBP2 expression by hepatitis C virus core protein

AIM: To investigate the molecular mechanism for regulation of cholesterol metabolism by hepatitis C virus(HCV) core protein in Hep G2 cells.METHODS: HCV genotype 1b core protein was cloned and expressed in Hep G2 cells. The cholesterol content was determined after transfection. The expression of sterol regulatory element binding protein 2(SREBP2) and the rate-limiting enzyme in cholesterol synthesis(HMGCR) was measured by quantitative real-time PCR and immunoblotting after transfection. The effects of core protein on the SREBP2 promoter and 3'-untranslated region were analyzed by luciferase assay. We used different target predictive algorithms, micro RNA(mi RNA) mimics/inhibitors, and site-directed mutation to identify a putative target of a particular mi RNA.RESULTS: HCV core protein expression in Hep G2 cells increased the total intracellular cholesterol level(4.05 ± 0.17 vs 6.47 ± 0.68, P = 0.001), and this increase corresponded to an increase in SREBP2 and HMGCR m RNA levels(P = 0.009 and 0.037, respectively) and protein expression. The molecular mechanism studyrevealed that the HCV core protein increased the expression of SREBP2 by enhancing its promoter activity(P = 0.004). In addition, mi R-185-5p expression was tightly regulated by the HCV core protein(P = 0.041). Moreover, overexpression of mi R-185-5p repressed the SREBP2 m RNA level(P = 0.022) and protein expression. In contrast, inhibition of mi R-185-5p caused upregulation of SREBP2 protein expression. mi R-185-5p was involved in the regulation of SREBP2 expression by HCV core protein. CONCLUSION: HCV core protein disturbs the cholesterol homeostasis in Hep G2 cells via the SREBP2 pathway; mi R-185-5p is involved in the regulation of SREBP2 by the core protein.

Genome-wide analysis of OVATE family proteins in cucumber(Cucumis sativus L.)

OVATE family proteins(OFPs)are plant-specific proteins with a conserved OVATE domain that regulate plant growth and development.Although OFPs have been studied in several species,their biological functions remain largely unknown in cucumber(Cucumis sativus L.).This study identified 19 Cs OFPs distributed on seven chromosomes in cucumber.Most Cs OFP genes were expressed in reproductive organs,but with different expression patterns.Ectopic expression of Cs OFP12-16c in Arabidopsis resulted in shorter and blunt siliques.The overall results indicated that Cs OFP12-16c regulates silique development in Arabidopsis and may have a similar function in cucumber.

Effects of XUEZHIKANG on Oxidized Low Density Lipoprotein,C - Reactive Protein, Fibrinogen in Unstable Angina Pectoris Patients

Objectives To study the effects of XUEZHIKANG on lipid modulating and the level of oxidized low density lipoprotein(OX - LDL), C -reactive protein(CRP), fibrinogen(FIB) in serum. Methods XUEZHIKANG was given to patients with unstable angina pectoris and hyperlipidemia at a dose of 0. 6 gram bid for 2 months and with half -dose for another 2 months. Vitamin E was given to unstable angina pectoris patients with normal lipid at the dose of 0. 1 gram bid for 4 months respectively. Then compared the level of lipid and OX - LDfL, CRP, FIB in serum at beginning, first - month and second -month. Results XUEZHIKANG can reduce the serum level of total cholesterol, low density lipoprotein in 1 month , and gained better effect in 2 months. It can also reduce triglyceride and increase high density lipoprotein in 2 months. Compared with vitamin E XUEZHIKANG can reduce the level of OX - LDL, CRP, FIB significantly after treatment for 2 months. Conclusions XUEZHIKANG has significant effect in lipid modulating , and it can also inhibit the development of inflammation in coronary plaque.

电针心经穴位对急性心肌缺血模型大鼠内侧隔核白细胞介素-2、JunB蛋白及c-fos表达水平的影响

目的观察电针心经对急性心肌缺血(acute myocardial ischemia,AMI)大鼠内侧隔核白细胞介素(interleukin,IL)-2水平、JunB蛋白及c-fos表达水平的影响,探究内侧隔核在针刺抗AMI中的作用及机制。方法将大鼠分为伪手术组、模型组和电针组,每组6只;结扎冠状动脉左前降支复制AMI大鼠模型;电针组大鼠选取手少阴心经“神门—通里”段进行干预,每次30 min,每日1次,连续电针3 d,刺激电流为1 mA,频率为2 Hz;伪手术组、模型组大鼠不进行电针干预。采用ELISA法检测大鼠大脑内侧隔核IL-2水平,Western blot法检测大鼠大脑内侧隔核区JunB蛋白表达水平,免疫荧光法检测大鼠大脑内侧隔核区c-fos免疫反应阳性神经元表达水平。结果与伪手术组比较,模型组大鼠大脑内侧隔核区IL-2、JunB蛋白水平显著升高(P<0.05),c-fos免疫反应阳性神经元数和平均吸光度(optical density,OD)值显著增加(P<0.05);与模型组比较,电针组大鼠大脑内侧隔核区IL-2、JunB蛋白水平显著降低(P<0.05),c-fos免疫反应阳性神经元数和平均OD值显著减少(P<0.05)。结论内侧隔核参与电针心经抗AMI的作用,其机制可能与电针降低大脑内侧隔核区IL-2、JunB蛋白及c-fos表达水平有关。

Exosomes derived from microglia overexpressing miR-124-3p alleviate neuronal endoplasmic reticulum stress damage after repetitive mild traumatic brain injury

We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.

桂皮醛对NIH3T3细胞c-Fos、c-Myc表达的影响

目的:研究肉桂主要成分桂皮醛刺激NIH3T3细胞后c-Fos、c-Myc蛋白在不同时点表达的规律,探讨桂皮醛促进NIH3T3细胞增殖的机制。方法:采用MTT法观察不同浓度桂皮醛对NIH3T3细胞增殖的影响;并采用免疫细胞化学法检测桂皮醛对NIH3T3细胞c-Fos、c-Myc蛋白表达的影响。结果:桂皮醛浓度在(8.8×10-2)-(8.8×10)μg/L浓度范围内对NIH3T3细胞具有促增殖作用。当其浓度为5.5μg/L时,促增殖作用最显著。在此浓度下,经桂皮醛刺激后,c-Fos和c-Myc蛋白均在2h开始表达,3h时表达明显增加。结论:桂皮醛可以上调c-Fos、c-Myc蛋白的表达,提示桂皮醛促进细胞增殖可能与其能促进c-Fos、c-Myc快速表达有关。

金雕肺的组织结构及KGF、c-Fos和Bax的表达

为了解金雕(Aguila chrysaetos)肺的组织结构特征,观察角质细胞生长因子(KGF)、c-Fos蛋白和Bax蛋白在肺中的表达情况,应用生物显微技术和免疫组织化学方法观察了金雕肺的组织结构,检测KGF、c-Fos和Bax蛋白在肺中的表达。结果表明,金雕的肺由各级支气管及其周围的呼吸毛细管组成,肺房呈辐射状开口于三级支气管周围,呼吸毛细管相互吻合成网状,周围有丰富的毛细血管网。KGF、c-Fos和Bax在三级支气管、肺房和呼吸毛细管的上皮细胞中呈免疫反应阳性,三者的表达强度有差异。KGF、c-Fos和Bax可能发挥不同的功能,它们的协同表达可能调控鸟肺细胞的生长、发育及成熟。

肠易激综合征大鼠内脏敏感性和脊髓背角5-HT和c-fos的表达

目的:探讨便秘型肠易激综合征(C-IBS)大鼠模型对直肠球囊扩张的内脏敏感性改变和脊髓背角5-羟色胺(5-HT)、c-fos的分布和异常表达.方法:C-IBS大鼠模型组应用冰水ig方法建立(A组,n=10),和正常对照大鼠(B组,n=10).所有大鼠给予直肠内球囊扩张,检测球囊扩张引起腹部收缩反射的最小容量阈值及球囊不同容量扩张时腹部收缩反射的次数.腰骶段脊髓背角5-HT和c-fos表达应用免疫组织化学染色及计算机图像分析系统半定量进行分析.结果:直肠内球囊扩张时,A组引起腹部收缩的最小容量阈值略高于B组,无明显统计学差并(0.59±0.09 vs 0.57±0.13,P>0.05).直肠球囊扩张体积1.0mL时A组腹部收缩反射次数低于B组(10.3±3.3 vs 18.3±5.5,P<0.05);体积1.5和2.0mL高容量扩张时两组无明显差异(P>0.05).A组腰骶段脊髓背角5-HT、c-fos阳性神经组织的面积均明显高于B组(5-HT面积:146.5±15.1 vs 109.3±18.5;5-HT OD:45826±2563.2 vs 29358±8965.5:c-fos面积:125.4±23.3 vs 88.7±23.2;c-fos OD:46258±4642 vs 33238±4587;均P<0.05).结论:C-IBS大鼠模型存在对直肠球囊扩张的内脏敏感性异常,脊髓背角5-HT、c-fos的异常表达可能参与C-IBS大鼠内脏敏感性异常的调节.

电针“内关"对急性心肌缺血大鼠心肌c-fos基因表达的影响

目的:探讨电针“内关”改善急性心肌缺血的机制。方法:将96只大鼠随机分为假手术组、心肌缺血模型组、心肌缺血模型加电针组,采用结扎冠状动脉左前降支建立急性心肌缺血实验模型,造模成功后,电针双侧“内关”。用生化方法检测血清心肌酶活性,逆转录聚合酶链反应(RT-PCR)方法检测心肌中c-fos基因的表达。结果:心肌缺血模型组血清心肌酶活性和心肌c-fosmR—NA表达显著高于假手术组(P<0.05),电针后降低,差异有显著性意义(P<0.05)。结论:电针“内关”改善急性心肌缺血的机制与下调心肌组织c—fosmRNA的表达有关。

氯化锂-匹罗卡品诱发大鼠癫痫持续状态后脑内c-fos蛋白表达的动态观察

目的在氯化锂-匹罗卡品诱发癫痫持续状态模型中观察c-fos蛋白表达的时程变化。方法采用氯化锂-匹罗卡品诱发癫痫持续状态大鼠模型,观察大鼠的行为改变,并用免疫组化方法观察癫痫持续状态后不同时间点脑内c-fos蛋白表达的变化。结果癫痫持续状态后6h,1d时脑内c-fos蛋白表达最高,3d时表达次之,7d时极少数表达并接近对照组表达水平。结论癫痫持续状态后脑内c-fos蛋白表达持续较长时间;在此时期应给予干预措施,以减低脑内神经元损伤可能。

灵芝孢子粉对癫痫大鼠脑组织IL-1β与c-Fos的影响

目的:观察和探讨灵芝孢子粉对癫痫(EP)大鼠脑组织白细胞介素-1β(IL-1β)和c-Fos含量变化的影响。方法:用放射免疫学方法检测IL-1β水平,用免疫组织化学方法检测c-Fos的表达。结果:灵芝孢子粉组脑皮质和海马区c-Fos阳性细胞数明显少于癫痫模型组动物,脑组织中IL-1β明显低于癫痫模型组动物。结论:灵芝孢子粉能够有效降低癫痫大鼠脑组织IL-1β水平,纠正免疫失调而起到抗癫痫作用;灵芝孢子粉能够抑制癫痫大鼠脑组织c-Fos的表达,阻断迟反应基因(LRG)以达到抗癫痫作用。

高氧、维甲酸对胎鼠肺成纤维细胞c-Jun、c-Fos表达的影响

目的 探讨维甲酸对高氧暴露下胎鼠肺成纤维细胞 c- Jun、c- Fos表达的影响。方法 取第 2代胎鼠肺成纤维细胞作体外培养 ,待生长至接近融合状态时 ,随机分为 :空气组 ,空气 +维甲酸组 ,高氧组 ,高氧 +维甲酸组。于培养30 m in和 1、 2、 6、 12、 2 4、 4 8、 72 h时 ,行细胞固定后 ,应用免疫组化方法检测 c- Jun、c- Fos表达强度并结合计算机图像处理系统进行分析。结果 与空气组相比 ,高氧 1h时 c- Jun表达明显增强 (P<0 .0 1) ,6～ 12 h时达高峰 ,以后开始下降 ,但仍高于正常水平 (P<0 .0 1) ;c- Fos的表达在高氧 30 m in即增强 (P<0 .0 1) ,2～ 6 h达高峰 ,以后开始下降 ,2 4 h后恢复到正常水平。维甲酸能明显下调高氧所诱导的 c- Jun、c- Fos高表达 ,但并不能完全阻止 c- Jun、c- Fos的过度表达。结论 c- Jun、c- Fos可能参与了高氧暴露下胎鼠肺成纤维细胞的信号转导过程 ;维甲酸可部分抑制由高氧所诱导的 c- Jun、 c-

胍丁胺抑制福尔马林诱导的脊髓PKCγ,pCREB,c-Fos和c-Jun表达上调

目的研究福尔马林致炎性疼痛脊髓蛋白激酶Cγ(PKCγ),磷酸化的环磷酸腺苷反应元件结合蛋白(pCREB),即刻早期基因c-fos和c-jun表达的变化及胍丁胺对其的影响。方法♂SD大鼠,180～220g,随机分为:①生理盐水组;②福尔马林组;③胍丁胺(160mg·kg-1,ip)组。足底注射5%福尔马林50μl后10、20、120min取L4,5脊髓,通过免疫组化和免疫印迹研究,检测胍丁胺对炎性疼痛脊髓PKCγ、pCREB、c-Fos和c-Jun蛋白表达变化的影响。结果大鼠单侧足底注射5%福尔马林10min后,引起双侧脊髓膜结合PKCγ表达量升高;足底注射20min后,双侧脊髓pCREB表达量升高;足底注射2h后,注射侧脊髓背角c-Fos和c-Jun蛋白表达量升高,注射对侧脊髓c-Fos和c-Jun蛋白表达量没有改变。胍丁胺明显抑制福尔马林所引起的膜结合PKCγ、pCREB、c-Fos和c-Jun蛋白表达量的升高(P<0.01)。结论炎性疼痛引起脊髓膜结合PKCγ、pCREB、c-Fos和c-Jun蛋白表达上调可能参与疼痛及痛觉过敏的产生。胍丁胺的镇痛机制可能与抑制PKCγ、pCREB、c-Fos和c-Jun等痛觉相关的信号转导分子的表达有关。

视觉刺激诱导c-Fos在弱视成年大鼠视皮质神经元表达的研究

目的:研究光刺激诱导双眼形觉剥夺性弱视成年大鼠视皮质神经元c-Fos表达情况。方法:缝合14d龄SD大鼠双侧眼睑建立双眼形觉剥夺性弱视模型,无光照弱视组8只,光照弱视组8只。饲养至100d龄时剪开眼睑,光照弱视组暴露于外界光线中0.5h,无光照弱视组不接触光线。取材后对两组视皮质组织进行Nissle染色和c-Fos蛋白免疫组化染色,并对染色结果进行计算机图像分析。结果:光照弱视组视皮质表达c-Fos蛋白阳性神经元显著多于无光照弱视组(P<0.05)。结论:双眼形觉剥夺弱视成年大鼠视皮质内经光刺激诱导c-Fos表达增加,提示已过视觉发育敏感期成年大鼠的视皮质仍存留一定程度的视觉可塑性。

连续及部分睡眠剥夺96小时后大鼠脑干中c-Fos蛋白的表达

目的 :探索连续及部分不同睡眠剥夺条件下的脑c -Fos蛋白的表达。方法 :采用小平台水环境法建立大鼠睡眠剥夺模型 ,用Fos蛋白免疫组化的方法分别测量连续睡眠剥夺 96小时组、部分睡眠剥夺 96小时组、大平台对照组和正常单独饲养组脑干中Fos蛋白的表达 ,每组 4只。结果 :同连续睡眠剥夺组相比 ,部分睡眠剥夺组表达范围更广 ,在脑干网状结构及中缝的被盖背侧核、脑桥被盖网状核及脑桥吻侧网状核、脑桥尾侧网状核、丘脑的中线核群、板内核群和网状核均有所表达 ,但阳性程度低。结论 :部分睡眠剥夺对脑的影响要小于连续睡眠剥夺。

老年抑郁大鼠海马CA3区c-fos和Caspase-3蛋白的表达

目的:探讨慢性应激老年抑郁大鼠海马CA3区c-fos和Caspase-3蛋白的表达,以探讨老年抑郁症的发病机制。方法:将20只Wistar老年大鼠随机分为实验组和对照组,每组10只。实验组大鼠每天强迫游泳20 min,并每周给予电刺激和强光照射各1次,经过21 d应激制作老年大鼠慢性应激抑郁模型,对照组正常喂养,不给予上述处理。采用特异性抗体标记的免疫组织化学方法,检测实验21 d后老年大鼠海马CA3区c-fos和Caspase-3蛋白的表达情况,并观察大鼠开场行为和排便情况。结果:①实验组老年大鼠中央格停留时间延长,水平穿越格数减少,修饰行为减少,排便次数增多;②c-fos平均灰度值:实验组(166.56±9.73),对照组(149.62±12.91),有显著差异P=0.000;③阳性细胞数:实验组(41.12±4.72),对照组(31.12±3.25),P=0.000;④caspase-3平均灰度值:实验组(202.59±2.64),对照组(179.74±8.24),有显著差异P=0.000;⑤阳性细胞数:实验组(24.73±2.57),对照组(18.86±1.13),有显著差异P=0.000。结论:慢性应激老年抑郁大鼠海马CA3区c-fos和caspase-3蛋白表达增加,行为表现异常,可能与慢性应激所致细胞凋亡增强,海马区脑组织损伤有关。

全脑暂时性缺血诱导c-fos原癌基因蛋白(FOS)在大鼠脑内的表达

用免疫组化(ABC)方法,普查了血管阻塞后全脑暂时性缺血刺激所诱导的c-fos原癌基因蛋白(FOS)在大鼠全脑各部的表达,观察了其分布、时间发展过程和变化。脑缺血再循环后15分钟至72小时内,在脑的各级水平的多数核团和结构出现不同程度的FOS表达。在不同的部位和功能区,FOS表达出现的时间、达到高峰时间和消退时间不同,表达的细胞多少和强度也不相同。在各脑室的室管膜细胞、触液神经元和下丘脑(特别是视上核和室旁核)最先出现,并呈现一过性的FOS快速表达。海马的FOS表达在2h后出现、主要局限在齿状回、下托和CA_4、CA_3区。在边缘系统和嗅脑的扣带皮质、梨状皮质、杏仁、隔核和内侧缰核、嗅球外颗粒层和僧帽层以及前嗅核呈现高水平的持久表达。缺血后2～48h内,新皮质Ⅱ—Ⅵ层诱导出广泛的高水平表达。丘脑、基底核、中脑的FOS表达则出现较晚且分布稀疏。FOS在脑干的表达因不同核团而异。本文结果提示:全脑暂时性缺血刺激后,脑内不同部位、不同核团和功能区激活FOS表达的机制不同,其功能意义也可能有所不同。

剥夺性弱视猫外侧膝状体c-fos基因及Fos蛋白表达

为研究生后关键期内,正常猫及剥夺性弱视猫外侧膝状体神经元功能形态的变化规律,对正常视发育敏感期的幼猫行左眼睑缝合术,造成剥夺性弱视模型,采用Nissl染色法及电镜观察正常猫及剥夺性弱视猫外侧膝状体神经元的形态改变,并用免疫组化和原位杂交技术检测Fos蛋白及cfos基因的表达情况。结果显示:(1)剥夺猫剥夺层外侧膝状体神经元的截面积比非剥夺层及正常猫的明显变小,差异有显著性(P<0.05);(2)超微结构显示:剥夺性弱视猫的剥夺层出现大量变性的细胞,剥夺4周时最为显著;(3)正常猫外侧膝状体神经元Fos蛋白和cfos原位杂交的阳性细胞数及OD值随着时间的延长而降低。同龄猫剥夺层外侧膝状体Fos蛋白和cfos原位杂交阳性细胞数及OD值较正常猫减少(P<0.05)。结果提示:猫视觉的可塑性敏感期为生后4周到8周。剥夺性弱视猫外侧膝状体剥夺层及非剥夺层神经元形态及功能与正常猫相比较均发生改变。Fos法可以成为衡量弱视猫外侧膝状体兴奋性状态与形态学定位相结合的实验方法,但必须注意神经可塑性对其的影响。