电针对束缚应激大鼠下丘脑神经元活化的调节作用

目的 观察应激大鼠下丘脑神经元的活化程度,以及电针足三里穴对它的调节作用。方法将大鼠随机分为对照组、束缚应激组、束缚应激加针刺组,以标志神经元活化的c fos蛋白作为检测指标,用Western印迹杂交法观察三组大鼠下丘脑内c fos蛋白的变化。结果 束缚应激大鼠下丘脑内c fos蛋白表达水平明显高于对照组(P<0. 01)。束缚应激大鼠经针刺足三里穴后下丘脑内c fos蛋白与应激组相比明显回落,在针刺结束后的各个时间点(0h、1h、3h)下丘脑内c fos蛋白均比应激组有所下降,其差异均具有显著性意义(P<0. 05)。结论 束缚应激能够引起下丘脑c fos蛋白的表达增加,针刺足三里穴能够下调下丘脑c fos蛋白的表达水平,针刺可能在预防应激性疾病中具有重要作用。     

慢性心理应激大鼠下丘脑环核苷酸和不同脑区核团c-fos蛋白表达及加味四逆散的影响

目的 观察慢性心理应激反应大鼠下丘脑环核苷酸系统和不同脑区核团c -fos蛋白表达的变化以及加味四逆散(JWSNS)的影响。方法 采用束缚制动的方法建立慢性心理应激反应大鼠模型,放射免疫法和免疫组织化学法检测大鼠下丘脑cAMP、cGMP含量和不同脑区核团c -fos蛋白表达的情况。结果 慢性心理应激可使大鼠下丘脑cAMP、cGMP含量显著升高 ( 1. 309±0. 62和 0. 136±0. 05 ) (P均 <0. 01),并可诱导c- fos在大鼠海马、大脑皮质、杏仁核、下丘脑室旁核(Pa)、室周核(Pe)以及弓状核(Arc)内广泛持续表达,其中以大脑皮质、杏仁核最为密集。JWSNS可明显降低心理应激大鼠下丘脑cAMP、cGMP含量(0. 877±0. 05和 0. 051±0. 02) (P均<0. 05)和上述脑区核团fos样免疫阳性神经元的数目,减少fos蛋白的表达。结论 cAMP、cGMP和c -fos参与慢性心理应激反应的机制;而JWSNS可能通过影响环核苷酸系统和c- fos蛋白的表达对慢性心理应激反应进行调控。     

大鼠蛛网膜下腔出血不同时相脑组织C-fos表达实验研究

目的：探讨大鼠蛛网膜下腔出血不同时相的症状及行为表现与脑组织C-fos表达的关系及其生物学意义.方法：选用Wistar大鼠70只,建立大鼠蛛网膜下腔出血模型,随机分为蛛网膜下腔出血30 min组、1 h组、2 h组、6 h组、24 h组、72 h组、7 d组,共7组,每组6只.对照组（生理盐水对照组）共7组,每组4只.取脑做连续切片后,采用HE染色,光镜下观察并进行定位.采用C-fos免疫组化染色,在显微镜下观察C-fos于不同时相在脑膜、脑皮质区及相关核团如尾壳核、苍白球、海马、背侧丘脑腹后外侧核、小细胞网状核、背侧旁巨细胞核的阳性细胞数.结果：对照组C-fos在尾壳核、苍白球、海马、背侧丘脑腹后外侧核、小细胞网状核、背侧旁巨细胞核表达极低;30 min、1 h、2 h、6 h组C-fos表达较高,与对照组比较差异均有统计学意义（P＜0.001）;24 h、72 h和7 d组C-fos表达较低,但是与对照组比较差异也均有统计学意义（P＜0.001）.结论：（1）大鼠蛛网膜下腔出血后不同时相C-fos在脑膜、脑皮质区及尾壳核、苍白球、海马、背侧丘脑腹后外侧核、小细胞网状核、背侧旁巨细胞核中都有表达且表达量随时相改变;（2）大鼠蛛网膜下腔出血后相关症状与不同时相C-fos在脑中表达之间存在着相关联系,具有规律性.     

腹腔神经节毁损对胃扩张兔脊髓c-fos表达的影响

目的:观察高强度聚焦超声(High Intensity Focused Ultrasound,HIFU)毁损腹腔神经节(Celiac ganglion,CG)对胃扩张兔脊髓c-fos表达的影响,探讨其治疗疼痛的可能机制.方法:健康成年新西兰大白兔20只,随机分为4组:非扩张非毁损组(nDnB)、扩张非毁损组(DnB)、扩张毁损组(DB)、非扩张毁损组(nDB),每组各5只.在无菌操作下暴露CG,DB、nDB组用HIFU毁损神经节,nDnB、DnB组假手术,术后7天,均在麻醉下安置胃扩张装置.安置后7天,nDB、nDnB组在麻醉下灌注固定取胸段6～8(T6～8)脊髓,DB和DnB组采用反复梯度扩张胃1h后,灌注固定取脊髓.用免疫组织化学方法检测脊髓前角和后角Fos样免疫反应阳性神经元(Fos-like immunoreactivity neuron,FLIN)计数的变化.结果:各组FLIN比较,毁损腹腔神经节和(或)扩张胃均不能使脊髓后角及前角FLIN明显地增加(P＞0.05).结论:HIFU毁损CG对胃扩张兔脊髓c-fos表达无明显影响,这与其它内脏神经毁损对伤害性刺激引起的脊髓c-fos表达的影响不同,可能是因为对胃伤害性信息的介导存在特殊的中枢机制.     

兔肝外胆道内脏感觉传导途径的实验研究

目的:通过动物造模观察与内脏痛相关核团中C-fos、一氧化氮合酶(NOS)的表达,探讨内脏感觉传导途径。方法:新西兰大白兔36只,随机分为假手术a组、假手术b组、胆管注射大肠杆菌组、胆囊内注射大肠杆菌组、胆管扩张组、扩张Oddis括约肌组共6组,每组6只,取脊髓、脑干以及间脑作连续切片后,采用HE染色光镜定位及免疫组织化学技术检测各组脑和脊髓不同部位C-fos、NOS的表达,并进行比较。结果:胆管注射大肠杆菌组、胆囊内注射大肠杆菌组在脊髓(C5-7、T6-9)后角、薄束核、楔束核、丘脑腹后外侧核、孤束核中NOS的阳性表达明显高于假手术a组(P<0.05);胆管扩张组脊髓(C5-7、T6-9)后角、薄束核、楔束核、孤束核、丘脑腹后外侧核中C-fos的阳性表达明显高于假手术b组(P<0.05);扩张Oddis括约肌组脊髓后角(C5-7)和孤束核中C-fos的阳性表达明显高于假手术b组(P<0.05)。结论:胆囊和胆管引起的疼痛均与脊髓(C5-7、T6-9)后角、薄束核、楔束核、丘脑腹后外侧核、孤束核有关,推测由交感和迷走神经双重支配;Oddis括约肌引起的疼痛与孤束核有关,推测由迷走神经传入。     

电针对血管性痴呆小鼠海马神经元c-fos表达的影响

目的观察电针对血管性痴呆(VD)小鼠海马神经元凋亡及相关基因cfos表达的影响。方法以反复脑缺血再灌注法复制VD小鼠模型,电针肾俞、膈俞和百会穴,对照组灌胃喜得镇,分别于治疗1d和7d后,以FCM法检测小鼠海马细胞凋亡率、cfos表达。结果术后1d,模型小鼠海马细胞凋亡率(6.86±1.10)较假手术组(3.59±0.70)明显增高(P<0.01),术后7d(13.49±1.72)较术后1d还高(P<0.01)。术后1d,模型小鼠海马细胞cfos表达(1.08±0.33)较假手术组(1.00±0.25)有升高趋势(P>0.05),7d时(2.65±0.29)则显著升高(与术后1d和假手术组比较,均P<0.01)。电针在两个时点均可上调小鼠海马细胞cfos的表达(2.26±0.31,3.76±0.41),降低细胞凋亡率(3.68±1.48,6.36±1.52),与模型组相比均差异有显著性(P<0.01)。结论电针具有抑制细胞凋亡、保护神经元的作用,这可能是电针改善VD小鼠智能的作用机理之一。     

Nitric oxide synthase-containing magnocellular neurons of the rat hypothalamus synthesize oxytocin and vasopressin and express Fos following stress stimuli

We investigated the chemical and anatomical features of nitric oxide synthase (NOS)-containing neurons in the paraventricular and supraoptic nuclei in the rat hypothalamus using combinations of enzyme histochemistry, in situ hybridization and immuno-histochemistry. Neurons expressing NOS mRNA completely overlapped with NADPH-diaphorase-positive neurons. Topographical distribution of NOS was segregated from that of CRF-containing parvicellular neurons in the posterior paraventricular nucleus but overlapped with that of magnocellular neurons. In the paraventricular nucleus, 70% of oxytocin neurons contained NOS, which corresponded to one half of NOS neurons. About one third of vasopressin-immunoreactive neurons were NADPH-diaphorase-positive and the same proportion of NADPH-diaphorase-positive neurons were vasopressin-immunoreactive. In the supraoptic nucleus, 50% of oxytocin neurons were NADPH-diaphorase-positive, which corresponded to 40% of NOS neurons. About 25% of vasopressin neurons were NADPH-diaphorase-positive, and 30% of NADPH-diaphorase-positive neurons were vasopressin-immunoreactive. When NADPH-diaphorase histochemistry was performed first, subsequent immunostaining was markedly perturbed. Using fluoro-gold as a retrograde tracer, 4% of NADPH-diaphorase-positive neurons were shown to contribute to the descending projection to the spinal cord. About 40%–50% of NADPH-diaphorase-positive neurons exhibited Fos immunoreactivity after injection of lipopolysaccharide or hypertonic saline, while only 10%–15% of these neurons expressed Fos in response to immobilization or pain. Endogenous NO may be involved in the regulation of magnocellular functions, especially when the internal environment is disturbed.     

Expression of the c-fos oncogene in chemically-induced mouse tumours and in human skin tumours.

Using a polyclonal antibody to fos oncoprotein and an immunofluorescent technique, we investigated expression of the fos oncogene in chemically-induced mouse tumours and human premalignant and malignant skin lesion. In the chemically induced tumours, the nuclei of almost all carcinoma cells stained uniformly with this antibody, while positive cells were observed in the outermost layers in the benign papillomas. In human tumours, a greater degree of nuclear staining was observed in cases of squamous cell carcinoma than in tissues from patients with Bowen's disease. Basal cell carcinoma and malignant melanoma with histological evidence of invasiveness of the tumour cells showed a higher expression of the fos gene product than that seen in histologically circumscribed tumour nests. Thus, a higher expression of the fos oncogene is closely related to the malignant progression of tumour cells, in particular, the extent of invasiveness.