核因子-κB对哮喘患者T淋巴细胞血红素氧合酶-1表达的调控

探讨核因子κB（NF－κB）对哮喘患者T淋巴细胞HO－1表达的转录调节机制。分离18例急性发作期哮喘患者外周血T淋巴细胞，并分成3组培养：对照组、加入NF－κB激动剂肿瘤坏死因子－α（TNF－α）组、同时加入TNF－α和NF－κB抑制剂二硫代氨基甲醇吡咯烷（PDTC）组。培养6h后留取细胞，用逆转录聚合酶链反应（RT－PCR）检测血红素氧合酶－1（HO－1）的mRNA。培养24h后留取细胞用Western印迹法检测HO－1的表达。发现TNF－α组T淋巴细胞HO－1蛋白和mRNA表达水平显著高于对照组（q=44.48、29.94,P均＜0.01)，而同时加入TNF－α和PDTC培养组T淋巴细胞HO－1蛋白和mRNA表达水平显著低于TNF－α组（q=43.23、27.99,P均＜0.01)。可见哮喘患者T淋巴细胞HO－1基因转录可能是通过激活NF－κB进行调控。T淋巴细胞NF－κB－HO氧化激活途径可能是哮喘的发病机制之一。     

雷公藤抑制哮喘豚鼠模型IL-5、GM-CSF 表达以及核因子-κB的作用

目的 探讨雷公藤对哮喘豚鼠模型IL－5、GM－CSF表达的影响及其机制。方法 采用卵蛋白（OA）致敏复制哮喘豚鼠模型,用原位杂交（ISH）就雷公藤甲素（TP）对其肺组织和外周血单个核细胞（PBMC）IL－5、GM－CSF mRNA表达的影响进行检测。并通过凝胶电泳迁移试验（EMSA）检测TP对伴刀豆蛋白（ConA)或佛波脂（PMA）刺激的人T细胞核因子－κB（NF-κB）的DNA结合活性的影响。结果 哮喘豚鼠肺组织和PBMC IL－5、GM－CSF mRNA表达显著高于正常组（P＜0．01）和TP处理组（P＜0．05或0．01）。ConA或PMA刺激可使T细胞NF－κB的DNA结合活性增强,活性增强的NF－κB为多克隆抗体P65和P50的异源二聚体,TP可以降低该NF－κB的DNA结合活性。结论 雷公藤通过抑制NF－κB的DNA结合活性,进而抑制IL－5、GM－CSF mRNA的转录,可能是其治疗哮喘的机制之一。     

内源性一氧化碳对低氧大鼠肺动脉转化生长因子-β3和Ⅰ型胶原表达的影响

目的:探讨内源性一氧化碳(CO)对低氧大鼠肺动脉胶原代谢的作用及其机制。方法:采用常压低氧大鼠肺动脉高压模型,观察血红素氧合酶(HO)抑制剂锌原卟啉-Ⅸ(ZnPP-Ⅸ)对肺动脉平均压(PAMP)和肺组织匀浆碳氧血红蛋白(HbCO)含量的影响,并用免疫组织化学和核酸原位杂交法分别观察ZnPP-Ⅸ对肺动脉转化生长因子-β₃(TGF-β₃)、Ⅰ型胶原蛋白的表达和肺动脉TGF-β₃mRNA、Ⅰ型前胶原mRNA和金属蛋白酶组织抑制因子(TIMP-1)mRNA表达的影响。结果:ZnPP-Ⅸ使低氧大鼠PAMP明显升高,肺组织匀浆CO含量明显降低;ZnPP-Ⅸ能促进TGF-β₃蛋白表达和TGF-β₃mRNA表达,显著促进低氧大鼠肺动脉Ⅰ型胶原蛋白表达和Ⅰ型前胶原mRNA表达,上调TIMP-1mRNA的表达。结论:内源性CO可能通过抑制TGF-β₃mRNA和TGF-β₃蛋白表达而抑制胶原蛋白的合成,促进胶原的降解,从而对低氧大鼠肺动脉胶原代谢发挥重要的调节作用。     

一氧化碳促进肺动脉平滑肌细胞血红素氧合酶基因的表达

目的:观察低浓度一氧化碳(CO)和低氧处理肺动脉平滑肌细胞(PASMC)后,血红素氧合酶(HO)基因表达的状况。方法:培养大鼠PASMC成功后并进行鉴定,应用免疫组织化学、逆转录聚合酶链式反应(RT-PCR)和Westernblot的方法研究低浓度CO和低氧作用后PASMC的HOmRNA及HO蛋白质含量。结果:①HO-1抗体免疫组织化学染色低氧组细胞胞浆呈黄色,中等强度;CO处理组的细胞胞浆呈棕黄色,强阳性,比低氧组的颜色显著深。HO-2抗体染色各组细胞胞浆均呈淡黄色,弱阳性。②低氧组细胞HO-1mRNA的表达水平为1.25±0.37,明显高于常氧组细胞(0.12±0.04);低浓度CO组细胞HO-1mRNA的表达水平为3.52±0.68显著高于低氧组。各组细胞HO-2mRNA的水平无明显差异。③低氧48h组细胞HO-1的蛋白含量为1.07±0.15,高于低氧24h组细胞(0.52±0.04);低浓度CO组细胞HO-1的蛋白含量为3.65±0.43,显著高于低氧组细胞,48h蛋白质含量为最高。结论:低浓度CO和低氧处理PASMC,可以促进HO-1mRNA和蛋白质的表达,而HO-1在细胞低氧损害中发挥调节作用,部分减轻低氧的损害。     

一氧化碳抑制缺氧所致大鼠肺动脉平滑肌细胞增殖

目的:研究内源性及外源性一氧化碳(CO)对缺氧状态下肺动脉平滑肌细胞(PASMC)增殖的影响。方法:选用第3-5代培养的大鼠PASMC,分别在常氧及缺氧条件下用HO酶促进剂氯血红素(hemin)、CO清除剂牛血红蛋白(Hb)和CO气体进行干预。通过MTT比色、增殖细胞核抗原(PCNA)蛋白的免疫细胞化学检测上述条件对PASMC代谢及增殖的影响,用流式细胞术检测不同条件对PASMC细胞周期的影响。结果:PASMC在缺氧时MTT比色、PCNA表达及S期、G₂/M期细胞所占比例明显高于对照组(P＜0.01),使用hemin和CO后上述指标显著低于缺氧组(P＜0.01或P＜0.05),而用Hb干预则PASMC代谢和增殖较缺氧组进一步增高(P＜0.01或P＜0.05)。结论:缺氧时PASMC产生的内源性CO以自分泌方式抑制PASMC自身增殖,使用hemin促进内源性CO生成或直接使用外源性CO气体均可抑制PASMC在缺氧状态下的增殖。     

低氧对肺动脉平滑肌和内皮细胞蛋白激酶C α mRNA表达的影响

目的：探讨低氧对肺动脉平滑肌和内皮细胞蛋白激酶C(PKC)αmRNA表达的影响。方法：应用原位杂交技术了大鼠不同节段肺动脉平滑肌细胞和内皮细胞PKCαmRNA的分布及低氧对在体和离体肺动脉平滑肌细胞及内皮细胞PKCαmRNA表达的影响。结果：正常大鼠各级肺动脉平滑肌细胞和内皮细胞均有PKCαmRNA的表达,腺泡内肺动脉平滑肌细胞中的表达明显高于肌型肺动脉（P＜0．01）,低氧14d和28d肌型动脉和腺泡内肺动脉内皮细胞的表达均明显增高（P＜0．01）,腺泡内腺动脉平滑肌细胞的表达较对照组明显升高（P＜0．01）,低氧14d肌型肺动脉平滑肌细胞表达升高不明显,但低氧28d明显升高（P＜01）,正常条件下离体培养猪肺动脉平滑肌细胞和内皮细胞均有PKCαmRNA的表达,低氧1h对其表达无明显影响,48,72h表达明显升高,以72h升高最显著（P＜0．001）,结论：低氧可促进肺动脉平滑肌细胞和内皮细胞PKC amRNA 的表达,而以腺泡内肺动脉平滑肌细胞的变化最明显,PCKα在低氧性肺动脉高压的形成中可能起一定作用。     

首发精神分裂症PBMCNF-κB活性及其mRNA表达与IL-1β、IL-6、TNF-αmRNA表达的关系

目的：探讨首发精神分裂症患者外周血单个核细胞（PBMC）NF-κB的活性及其mRNA表达与IL-β、IL-6、TNF-α mRNA表达的相互关系，为临床应用NF-κB调节剂提供理论依据。方法：应用NF-κB活性ELISA试剂盒，检测精神分裂症组和健康对照组PBMC中NF-κB活性，并采用RT-PCR方法，测定两组PBMC中NF-κB mRNA表达及IL-1β、IL-6、TNF-α mRNA表达。结果：①精神分裂症组PBMC中NF-κB的活性及NF-κB mRNA表达量与对照组相比均明显增高，差异有显著性（P〈0．05）；②精神分裂症组PBMC中IL-1β、IL-6、TNF-α mRNA表达量与对照组相比均明显增高，差异有显著性（均P〈0．05）；③在精神分裂症组及对照组中，PBMC中NF-κB的活性与其mRNA表达量无明显相关（均P〉0．05）；④在精神分裂症组和对照组中，PBMC中NF-κB的活性与IL-1β、TNF-α mRNA均呈正相关（均P〈0．05）；无论是在精神分裂症组或对照组中，PBMC中NF-κB的活性与IL-6 mRNA均无明显相关（均P〉0．05）。结论：精神分裂症患者PBMC中NF-κB mRNA表达及活性均增高，对NF-κB活性的检测更能反映NF-κB的转录调控功能情况；活化的NF-κB对IL-1β、TNF-α的基因转录起了重要的调控作用。     

低氧诱导因子1在低氧致肺动脉平滑肌细胞增殖中的作用

目的: 探讨低氧对肺动脉平滑肌细胞(PASMC)增殖和凋亡的影响,以及HIF-1α、P-ERK1/2、iNOS蛋白表达变化在其中的作用与意义。方法: 体外培养大鼠PASMC，设计常氧组、低氧组及ADM、L-NAME、PD98059干预组，用MTT比色法和PCNA的免疫组化法测定细胞增殖反应，用流式细胞仪检测细胞凋亡，用Western blotting法检测HIF-1α、P-ERK1/2、iNOS的蛋白表达。结果: （1）低氧24 h组的A值明显高于常氧组(P<0.01)，而PD98059及ADM干预组明显低于低氧组(P<0.01)， L-NAME干预组明显高于低氧组和常氧组(P<0.01)。（2）免疫组化表明，低氧24 h组呈阳性表达(P<0.01)。PD98059、ADM抑制了PCNA的表达(P<0.01)， L-NAME促进了PCNA的表达(P<0.01)。（3）各组在低氧培养24 h后，凋亡指数差异无显著(均P＞0.05)。（4）Western blotting表明常氧组少量HIF-1α、iNOS、 P-ERK1/2表达，低氧4 h后均表达增高(P<0.01),8 h仍维持在高峰(P<0.01)，而HIF-1α、P-ERK1/2在低氧24 h后表达下调。L-NAME促进了HIF-1α表达(P<0.01),PD98059部分抑制了HIF-1α、iNOS及P-ERK1/2表达(P<0.01)；ADM部分抑制了HIF-1α表达，促进iNOS表达(P<0.01)。结论: 低氧能促进肺动脉平滑肌细胞增殖，对细胞的凋亡无影响；HIF-1在低氧诱导肺动脉平滑肌细胞增殖中起重要作用。     

γ射线辐射对大鼠胸主动脉平滑肌细胞肾上腺髓质素和内皮素生成的影响

为观察^60COγ射线辐射对培养的大鼠血管平滑肌细胞（VSMC）肾上腺髓质素（Adm)内皮素（ET）生成的影响，以探讨辐射防治再狭窄的机制，采用逆转录－竞争性聚合酶链反应观察经0,1,,24,25Gy辐射的培养大鼠血管平滑肌细胞Adm和ET mRNA水平；用放免方法测定相应的Adm和ET的生成。发现1Gy辐射对VSMC Adm和ET的生成及基因表达均无显著影响。14Gy辐射后VSMC生成的Adm较对照组增加270％（P＜0．05），ET的生成较对照组降低27．3％（P＜0．01）；25Gy辐射使VSMC生成的Adm和ET含量分别增加233％（P＜0．05）和降低58．0％（P＜0．01）。14和25Gy辐射分别使细胞Adm mRNA水平较对照组增加82．4％（P＜0．01）和101％（P＜0．01），使ET mRNA水平较对照组降低47．1％（P＜0．01）及40．2％（P＜0．01）。表明^60Coγ射线辐射可以促进Adm的蛋白及基因表达，而抑制ET的蛋白及基因表达，增加了Adm/ET比值。     

低分子肝素对缺血再灌注大鼠肾组织核因子-κB表达的影响

探讨低分子肝素对缺血再灌注大鼠肾组织核因子-κB（NF-κB）表达的影响。建立大鼠IRI模型，健康WistaI大鼠80只随机分为正常对照组、假手术组、模型未治疗组、LMWH治疗组，后两组又分别分为术后1、3、6、24h组。检测各组血清肌酐（Scr）水平及中性粒细胞（PMNs）细胞间黏附分子-1（ICAM-1）表达；通过光镜和免疫组织化学方法观察各组大鼠肾组织形态学及趋化因子NF-κB表达变化。结果表明：（1）肾缺血再灌注未治疗组造模后1h，Scr水平虽然没有明显变化，但ICAM-1、NF-κB表达增多，肾小管坏死积分值亦较假手术组明显增加（P〈0．01）；（2）缺血再灌注6h以后，两组Scr浓度明显增高（P〈0．01），但LMWH治疗组SCr、ICAM-1、NF-κB表达水平及肾小管坏死积分值均明显低于模型未治疗组（P〈0．05）；（3）肾组织中NF-κB表达与肾小管损伤积分值呈现良好的相关性（r=0．71，P〈0、01）；而NF-κB与ICAM-1间则呈现显著正相关（r=0．62，P〈0．05）。由此说明：（1）ICAM-1、NF-κB在肾缺血再灌注早期的瞬时表达，可能参与了炎症早期的白细胞迁移与浸润，与肾损伤的发生密切相关；（2）LMWH可通过减少ICAM-1及NF-κB的表达，阻抑炎症反应过程，减轻肾组织损伤。     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.     

Counterpointing the functional role of the forebrain and of the brainstem in the control of the sleep-waking system

This paper reviews the lifetime contributions of the author to the field of sleep-wakefulness (S-W), reinterprets results of the early studies, and suggests new conclusions and perspectives. Long-term cats with mesencephalic transection show behavioral/polygraphic rapid eye movement sleep (REMS), including the typical oculo-pupillary behavior, even when the section is performed in kittens prior to S-W maturation. REMS can be induced as a reflex. Typical non-rapid eye movement S (NREMS) is absent and full W/arousal is present only after a precollicular section. The isolated forebrain (IF) rostral to the transection exhibits all features of W/arousal and NREMS [with electroencephalographic (EEG) spindles and delta waves], arousal to olfactory stimuli, and including the appropriate oculo-pupillary behaviors. These features also mature normally after neonatal transection. REMS is absent from the IF. After deprivation there is NREMS pressure and rebound in the IF, but the decerebrate cat only shows pressure for REMS. Most IF reactions to pharmacologic agents are within expectations, except for the tolerance/withdrawal effects of long-term morphine use which are absent. In contrast, these effects are supported by the brainstem (i.e. seen in the decerebrate cat). In cats with ablation of the telencephalon, or diencephalic cats, delta waves are absent in the thalamus. EEG thalamic spindle waves are seen triggering S for only 4-5 days after ablation. Therefore, true NREMS is absent in chronic diencephalic cats although pre- and postsomniac behaviors persist. These animals are hyperactive and show a pronounced, permanent insomnia; however, a low dose of barbiturate triggers a dramatic REMS/atypical NREMS rebound. Cats without the thalamus (athalamic cats), initially show a dissociation between behavioral hyperactivity/insomnia and the neocortical EEG, which for 15-20 days exhibits only delta and slower oscillations. Fast, low-voltage W rhythms appear later on, first during REMS, but spindle waves and S postures are absent from the start, such that these cats also display only atypical NREMS. Athalamic cats also show barbiturate-sensitive insomnia. Cats with ablation of the frontal cortices or the caudate nuclei remain permanently hyperactive. They also show a mild, but significant hyposomnia, which is permanent in afrontal cats, but lasts for about a month in acaudates. The polygraphic/behavioral features of their S-W states remain normal. We conclude and propose that: (a) the control of the S-W system is highly complex and distributed, but is organized hierarchically in a well-defined rostro-caudal manner; the rostral-most or highest level (telencephalon), is the most functionally complex/adaptative and regulates the lower levels; the diencephalic/basal forebrain, or middle level, has a pivotal role in inducing switching between S and W and in coordinating the lowest (brainstem) and highest levels; (b) W can occur independently in both the forebrain and brainstem, but true NREMS- and REMS-generating mechanisms exist exclusively in the forebrain and brainstem, respectively; (c) forebrain and brainstem S-W processes can operate independently from each other and are preprogrammed at birth; this helps understanding normal and abnormal polygraphic/behavioral dissociations in humans and normal dissociations/splitting in aquatic mammals; (d) NREMS homeostasis is present in the IF, but only REMS pressure after deprivation persists in the decerebrate cat; (e) the thalamus engages in both NREMS and W; (f) insomnia in diencephalic cats is the result of an imbalance between antagonistic W- and S-promoting cellular groups in the ventral brain (normally modulated by the telencephalon); (g) the EEG waves, which are signature for each S-W state, appear to truly drive the concomitant behaviors, e.g. a hypothetical human IF could alternate between behavioral NREMS and W/arousal/awareness; (h) a role for REMS is to keep the individual sleeping at the end of the self-limiting NREMS periods. The need for accelerating research on telencephaling NREMS periods. The need for accelerating research on telencephalic S-W processes and downstream control of the lower S-W system levels is emphasized.     

Ultrastructure of the endothelium of the drainage system of the eye

The endothelium of the ocular drainage system (Schlemm’s canal, collector tubules, and aqueous veins) in primary juvenile glaucoma undergoes degenerative dystrophic changes with compensatory hypertrophy and proliferation at the initial stages of the glaucomatous process and atrophy and desquamation at advanced and terminal stages. Progressive decrease in the pinocytous function of endotheliocytes, reduction of the protein-synthesizing and mitochondrial compartments of the cytoplasm, and formation of autophagosomes reflect the process of endotheliocyte degeneration in general. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 574–577, May, 2008     

The development of the ligament of the head of the femur

The hip joints of 30 human male and female fetuses and stillborns between 20 mm and 350 mm crown-rump length were studied by light microscopy. The ligament of the head of the femur developed in situ as a condensation of mesenchyme at the end of the second month of intra-uterine life (IUL), and was vascularized by branches of acetabular vessels early in the fourth month. In the majority of fetuses older than 5.5 months IUL, vessels in the ligament passed a short way into the femoral head within cartilage canals, to supply a small region around the fovea capitis. The remainder of the head was supplied by vessels in canals from around the upper part of the neck. The ligament changed from predominantly cellular to fibrous during the last 4 months of IUL. This increase in strength suggested significant mechanical functions in utero: limitation of adduction-flexion and opposition to postero-superior dislocation were the most likely.     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Involvement of the striatum in the central regulation of the hormonal functions of the gonads

Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms. Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 85, No. 4, pp. 594–597, April, 1999.