参附注射液对大鼠全脑缺血再灌注时核转录因子-κB表达的影响

目的探讨参附注射液（Shenfu Injection）对大鼠全脑缺血再灌注时核转录因子-κB（nuclearfac—torkappaB，NF-κB）表达的影响及其治疗作用。方法采用四血管阻塞的方法复制出大鼠全脑缺血再灌注模型，采用免疫组化法、原位杂交法和原位末端标记法分别检测假手术组（A组）、全脑缺血再灌注组（B组）、参附注射液治疗组（C组）海马CA1区NF-κB、肿瘤坏死因子-amRNA（TNF-amRNA）表达及细胞凋亡数的变化。结果与假手术组比较，全脑缺血再灌注组海马CAl区NF-κB和TNF-amRNA的表达及细胞凋亡数明显增加（P〈0．01）；NF-κB和TNF-amRNA的表达分别于再灌注6h和12h达到高峰，并持续到48h；细胞凋亡数随再灌注时间的延长而逐渐增多（P〈0．01）。参附注射液治疗后NF-κB和TNF-amRNA的表达下降，细胞凋亡数减少（P〈0．01）。结论在脑缺血再灌注救治过程中参附注射液可能通过抑制NF-κB与TNF-amRNA的表达，进而减少细胞凋亡而发挥脑保护作用。     

α-硫辛酸对大鼠脑缺血再灌注损伤炎性机制的影响

目的探讨α-硫辛酸对大鼠脑缺血再灌注损伤炎性反应机制的影响。方法将SD大鼠随机分为假手术组(A)、脑缺血再灌注组(B)和硫辛酸预干预组(C),应用线栓法制备大脑中动脉闭塞2 h后再灌注模型,分别于再灌注6 h、24 h、3 d、7 d采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑梗死灶大小,采用免疫组织化学和原位杂交的方法检测大鼠核转录因子(nuclear factor kappa B,NF-κB)、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的表达。结果与假手术组比较,模型组NF-κB的表达于再灌注6 h即增高(P<0.05),TNF-α的表达于24 h增高(P<0.05),3 d时均达高峰(均P<0.05),7 d时仍高于假手术组(均P<0.05);与模型组比较,硫辛酸预干预组于脑缺血再灌注24 h、3 d、7 d时NF-κB和TNF-α表达降低;α-硫辛酸预干预组NF-κB各时点组内比较未见统计学差异(F=2.245,P>0.05)。与模型组比较,α-硫辛酸预干预组梗死体积明显减小,神经功能缺损评分降低(P<0.05)。结论α-硫辛酸对大鼠脑缺血再灌注损伤具有一定的保护作用,其机制可能是α-硫辛酸抑制NF-κB、TNF-α的表达,从而抑制大鼠脑缺血再灌注损伤后的炎症反应有关。     

NF-κB信号通路在小鼠脑缺血再灌注细胞凋亡中的作用

目的初探核因子-κB(nuclear factor-kappa B,NF-κB)信号通路在小鼠脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)细胞凋亡中的作用。方法通过夹闭小鼠双侧颈总动脉建立动物模型。模型组分为3 h、6 h、12 h、1 d、3 d、7 d、14 d、21 d,共8组。TTC染色确定脑缺血损伤区域,TUNEL法检测不同时间缺血区细胞凋亡数,原位杂交法和Western blotting检测NF-κB信号通路靶基因/蛋白Bcl-2和C-myc mRNA及蛋白表达。PDTC抑制NF-κB信号通路后,TUNEL法和原位杂交检测建模后1 d、7 d、14 d神经细胞凋亡数及Bcl-2和C-myc mRNA变化情况。结果各模型组海马CA3区凋亡细胞数量、Bcl-2 mRNA和C-myc mRNA阳性细胞数及二者蛋白表达量高于正常组和假手术组(P0.05);PDTC抑制NF-κB信号通路后,各抑制组与对应时间点模型组相比,Bcl-2 mRNA阳性细胞数减少,TUNEL阳性细胞数和C-myc mRNA阳性细胞数增加(P0.05)。结论 CIRI早期即出现有凋亡细胞数的增加,并在其后较长一段时间内细胞凋亡过程仍存在着持续性的进展;NF-κB信号通路在CIRI病程进展中对神经细胞凋亡起抑制作用,其机制可能通过Bcl-2诱导和C-myc抑制共同发挥调节作用。     

雌激素对大鼠脑缺血再灌注损伤脑组织NF—κB、iNoS表达和细胞凋亡的影响

目的 研究雌激素对大鼠局灶性脑缺血再灌注(I/R)损伤后NF-κB、iNOS蛋白和细胞凋亡变化的影响.方法 采用线栓法复制局灶性脑I/R模型.应用免疫组化检测NF-κB、iNOS蛋白表达;利用原位缺口末端标记法(TUNEL )研究凋亡细胞的变化.结果 与对照组大鼠相比,雌激素组脑缺血再灌注后NF-κB、iNOS表达明显减弱(P<0.01);凋亡细胞显著减少(P<0.01).结论 雌激素能抑制脑I/R后NF-κB、iNOS表达,减少细胞凋亡,这可能是其脑保护作用的机制之一.     

长春西汀对大鼠脑缺血再灌注损伤炎性反应机制的影响

目的探讨长春西汀对大鼠脑缺血再灌注损伤炎性反应机制的影响。方法将Wistar大鼠随机分为假手术组、脑缺血再灌注损伤组和长春西汀干预组,应用线栓法制备大脑中动脉闭塞2h后再灌注模型,分别于再灌注6h、24h、3d、7d采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑梗死灶大小,采用干、湿重法检测脑组织含水量以评价脑水肿的程度,采用免疫组织化学和原位杂交的方法检测大鼠核转录因子(nuclear factor-kappa B,NF-κB)、肿瘤坏死因子α(TNF-α)的表达。结果与假手术组比较,模型组NF-κB的表达于再灌注6h即增高(P<0.05),TNF-α的表达于24h增高(P<0.05),3d时均达高峰(均P<0.05),7d时仍高于假手术组(均P<0.05);与模型组比较,长春西汀干预组于再灌注后24h、3d、7d时NF-κB和TNF-α表达降低。长春西汀干预组NF-κB各时点组内比较未见统计学差异(F=2.324,P>0.05)。与假手术组比较,模型组和长春西汀干预组脑组织水含量于脑缺血再灌注损伤6h即增高,3d时达到高峰(均P<0.05),7d时仍高于假手术组(均P<0.05)。结论长春西汀对大鼠脑缺血再灌注损伤炎性反应有明显的抑制作用。     

肠道菌群失调加重小鼠局灶性脑缺血再灌注后炎性反应

目的探讨肠道菌群失调对小鼠局灶性脑缺血再灌注后炎性反应的影响。方法小鼠随机分为假手术组、缺血再灌注组(对照组)和菌群失调组。假手术组和对照组每天胃内注射同体积生理盐水;菌群失调组胃内注射头孢曲松钠,均2次/d,共持续4 d。第5天,行脑缺血再灌注手术,术后72 h,Zea Longa法对小鼠进行神经功能评分,干湿重法测量缺血侧脑组织含水量; RT-PCR法检测缺血侧半球IL-6 mRNA和TNF-αmRNA表达;采用Western blot法检测缺血侧半球NF-κB和ZO-1的表达。结果缺血再灌注72 h后,与对照组比较,菌群失调组小鼠神经功能评分较高,缺血侧半球脑含水量、IL-6、TNF-αmRNA和NF-κB表达显著增加,ZO-1表达显著减少,差异均有统计学意义(P 0. 05)。结论肠道菌群失调加重了小鼠局灶性脑缺血再灌注损伤后炎性反应。     

大鼠局灶性脑缺血再灌注后Ref-1、NF-κB及IκB的表达

目的研究大鼠局灶性脑缺血再灌注损伤后Ref-1蛋白、NF-κB及IκB的动态变化。方法采用大脑中动脉线栓法制作局灶性脑缺血再灌注模型,应用免疫组化法检测Ref-1、NF-κB、IκB的动态表达。结果与对照组及假手术组相比,脑缺血再灌注后,随着再灌注时间的延长,Ref-1蛋白、IκB阳性细胞数逐渐减少(P<0.05),NF-κB蛋白阳性细胞数逐渐增加(P<0.05)。结论脑缺血区再灌注损伤可导致Ref-1蛋白酶活性下降、IκB降解,引起NF-κB的激活,进一步导致自由基大量产生,加重了细胞内DNA的损伤,从而促进损伤区的神经元凋亡。     

脑组织TOLL样受体2的激活与大鼠局灶性脑缺血再灌注损伤的关系

目的 探讨TOLL样受体2(TLR2)的激活与脑组织缺血再灌注损伤的关系.方法 线栓法制造局灶性脑缺血再灌注损伤大鼠模型,采用免疫组织化学法观察TLR2和核因子-κB(NF-κB)在不同再灌注时间点缺血脑组织的分布和表达量,同时应用酶联免疫方法检测肿瘤坏死因子-α(TNF-α)在不同再灌注时间点缺血脑组织内浓度,并分析其表达相关性.结果 大鼠局灶性脑缺血再灌注损伤过程中,TLR2蛋白在缺血半影区及皮质区、纹状体区有表达,且在灌注早期(即1h)即有增强表达,在3 h即达高峰;皮质区和纹状体区NF-κB表达从6 h开始逐渐增强,于再灌注24 h达高峰;缺血侧半球皮质内TNF-α变化规律与NF-κB相似,但表达高峰在再灌注12 h;TLR2蛋白的表达与NF-κB和TNF-α表达呈正相关.结论 TLR2在脑缺血再灌注过程中被激活,可能通过NF-κB导致炎性因子TNF-α等的大量表达介导参与了脑缺血再灌注损伤的过程.     

美满霉素抑制血管性认知功能损伤大鼠海马星型胶质细胞激活和神经炎症(英文)

目的观察美满霉素(minocycline)对血管性认知功能损伤大鼠海马组织GFAP、COX-2、NF-κB、IL-1β和TNF-α表达的影响,探讨美满霉素对血管性认知功能损伤脑保护作用的机制。方法Wistar大鼠随机分为假手术组(S组)、血管性认知功能损伤模型组(M组)和美满霉素治疗组(MT组)。免疫组织化学法检测大鼠海马组织COX-2和NF-κB的表达,蛋白质印迹和免疫组织化学法检测大鼠海马组织GFAP的表达,ELISA法检测大鼠海马组织IL-1β和TNF-α的表达。结果MT 组 GFAP、COX-2、NF-κB、IL-1β和 TNF-α表达较 M 组均降低(P<0.01) ;MT 和 M 组GFAP、COX-2、NF-κB、IL-1β和 TNF-α表达均显著高于 S 组(P<0.01)。结论美满霉素能降低血管性认知功能损伤大鼠海马组织中GFAP、COX-2、NF-κB、IL-1β和TNF-α的表达,抑制血管性认知功能损伤大鼠海马星型胶质细胞活化和神经炎症,发挥脑保护作用。     

1,25(OH)_2D_3减轻小鼠局灶性脑缺血再灌注后炎性反应

目的探讨1,25(OH)_2D_3对小鼠局灶性脑缺血再灌注后炎性反应的作用及其机制。方法造模前,通过一个月低维生素D饮食喂养,小鼠随机分为假手术组、局部缺血再灌注组(模型组)和1,25(OH)_2D_3组(治疗组)。造模前3 d始,假手术组和模型组每天腹腔注射2.4%乙醇,治疗组腹腔注射1,25(OH)_2D_3,共持续6 d。再灌注72 h后,Zea Longa法对鼠进行神经功能评分,干湿重法测量缺血侧脑组织含水量,RT-PCR法检测缺血侧半球IL-1βmRNA和TNF-αmRNA表达,采用Western blot法检测缺血侧半球NF-κB p65和Claudin-5的表达。结果与模型组比较,缺血再灌注后72 h,治疗组小鼠神经功能评分较低,缺血侧半球脑含水量、IL-1βmRNA、TNF-αmRNA和NF-κB p65表达显著减少,Claudin-5表达显著增加,差异均有统计学意义(P0.05)。结论 1,25(OH)_2D_3减轻小鼠局灶性脑缺血再灌注损伤后炎性反应,其机制通过抑制NF-κB的活化有关。     

Denervation study of synapses of organ of corti of old world monkeys

Fine structural characteristics of synapses in the spiral organ of Corti were examined, with reference to differences between inner and outer haircell systems, and to location of neurons of origin of efferent axons. Surgical interruption of crossed olivocochlear bundle, of vestibular nerve, of facial nerve, and excision of superior cervical ganglia were used to determine the pathways of efferent axons. Interruption of the vestibular nerve near the brainstem results in degeneration of all efferent terminals on outer hair cells. Mid-line lesions at, and caudal to, the facial colliculus result in degeneration of about half of these efferent terminals. Efferent synaptic bulbs to the inner hair-cell system are small, of the order of one micron, and form type 2 junctions with afferent dendrites. They tend to have more large dense-core vesicles (about 80 nm) than the large efferent terminals of the outer hair-cell system, and appear to be the terminals of axons in the habenula perforata, which exhibit varicosities laden with large dense core vesicles. The varicosities are unaffected by excision of the superior cervical ganglia. So far as our material can reveal, it appears that the varicosities in the habenula perforata do not survive vestibular root interruption, nor do the efferent processes in the internal spiral bundle or at the base of inner hair cells. Most interestingly, the afferent processes of the inner hair-cell system, as identified for example by their relation to pre-synaptic bodies in the inner hair cells, are subject to a trans-synaptic reaction after severance of the vestibular root. They undergo a dramatic cytological transformation, characterized by increase of volume, engorgement with microtubules, microfilaments, microvesicles of various sizes, and clusters of lysosomes. Thus, both the efferent and afferent terminals of the inner hair-cell system show marked cytological differences from the corresponding terminals of the outer hair cell system.     

Mechanism of action of tubocurarine on nicotinic cholinoreceptors of neurons of the sympathetic ganglia of rats

Tubocurarine (Tc) effect on membrane currents elicited by acetylcholine (ACh) was studied in isolated superior cervical ganglion neurons of rat using patch-clamp method in the whole-cell recording mode. The "use-dependent" block of ACh current by Tc was revealed in the experiments with ACh applications, indicating that Tc blocked the channels opened by ACh. Mean lifetime of Tc-open channel complex, tau, was found to be 9.8 +/- 0.5 s (n = 7) at -50 mV and 20-24 degrees C. tau exponentially increased with membrane hyperpolarization (e-fold change in tau corresponded to the membrane potential shift by 61 mV). Inhibition of the ACh-induced current by Tc (3-30 microM/1) was completely abolished by membrane depolarization to the level of 80-100 mV. Inhibition of ACh-induced current was augmented at increased ACh doses. It is concluded that the open channel block produced by Tc is likely to be the only mechanism for Tc action on nicotinic acetylcholine receptors in superior cervical ganglion neurons of rat.     

The effect of age of onset of PD on risk of dementia

Background Dementia occurs in the majority of patients with Parkinson’s disease (PD). Late onset of PD has been reported to be associated with a higher risk for dementia. However, age at onset (AAO) and age at baseline assessment are often correlated. The aim of this study was to explore whether AAO of PD symptoms is a risk factor for dementia independent of the general effect of age. Methods Two community-based studies of PD in New York (n = 281) and Rogaland county, Norway (n = 227) and two population-based groups of healthy elderly from New York (n = 180) and Odense, Denmark (n = 2414) were followed prospectively for 3–4 years and assessed for dementia according to DSM-IIIR. All PD and control cases underwent neurological examination and were followed with neurological and neuropsychological assessments. We used Cox proportional hazards regression based on three different time scales to explore the effect of AAO of PD on risk of dementia, adjusting for age at baseline and other demographic and clinical variables. Findings In both PD groups and in the pooled analyses, there was a significant effect of age at baseline assessment on the time to develop dementia, but there was no effect of AAO independent of age itself. Consistent with these results, there was no increased relative effect of age on the time to develop dementia in PD cases compared with controls. Interpretation This study shows that it is the general effect of age, rather than AAO that is associated with incident dementia in subjects with PD. Received in revised form: 22 December 2005     

Limits of deinstitutionalization--perspectives of relatives of psychiatric patients

After a hopeful beginning, the social process of the reintegration of those with severe mental illness has come to a standstill. I am led to wonder whether "the community" really wants to live together with people suffering from severe mental illness, and if so, how closely? As long as the medical treatment of mental illness provided by the general practitioners is fundamentally deficient, as they are not able to prescribe the necessary interventions--such as out-patient psychiatric nursing, and service providers in the out-patient sector are content with offering increasingly intensive forms of care for the less seriously ill at the cost of the Social Welfare System--the reintegration of those with serious mental illness remains an illusion--which is mainly to the benefit of providers of residential care in homes and hostels.