自噬水平变化对心脏骤停心肺复苏后大鼠海马神经元凋亡的影响

目的探讨自噬水平变化对心脏骤停心肺复苏(CA/CPR)后大鼠海马神经元凋亡的影响。方法将40只大鼠随机分为假手术组(sham)、CA/CPR模型组(model)、雷帕霉素(Rapa)组(CA/CPR+Rapa)及3-甲基腺嘌呤(3-MA)组(CA/CPR+3-MA)。采用呼气末夹闭气管窒息法复制大鼠CA/CPR动物模型,分别给予自噬激动剂Rapa 0.2 mg/kg及自噬抑制剂3-MA 10 mg/kg进行干预。采用神经功能缺陷评分(NDS)评价CA/CPR大鼠神经功能;用TUNEL染色法检测大鼠海马神经元的凋亡变化;用RT-PCR和Western blotting法检测大鼠海马内微管蛋白轻链3(LC3)、Beclin-1、Bax、Bcl-2及Caspase-3 mRNA和蛋白的表达水平。结果与假手术组比较,CA/CPR模型组大鼠NDS评分明显降低;海马神经元TUNEL染色阳性细胞数明显增多,凋亡率显著升高;海马内LC3、Beclin-1、Caspase-3、Bax表达上调,Bcl-2表达下调(P<0.05,P<0.01)。与模型组比较,CA/CPR+Rapa大鼠NDS评分明显降低,而海马神经元凋亡率明显有所增加,海马内LC3、Beclin-1、Caspase-3、Bax表达明显上调,而Bcl-2表达则明显有所下降;CA/CPR+3-MA大鼠NDS评分明显升高,而海马神经元凋亡率下降,海马内LC3、Beclin-1、Caspase-3、Bax表达明显下调,而Bcl-2表达则有所升高(P<0.05,P<0.01)。结论 CA/CPR后自噬水平升高促进海马神经元凋亡,自噬水平降低抑制海马神经凋亡,两者相互作用共同参与CA/CPR的病理过程。     

穴位埋线对血管性痴呆大鼠海马CA1区神经细胞凋亡的影响

目的 观察穴位埋线疗法对血管性痴呆（VD）大鼠海马CA1区神经细胞凋亡和Bcl-2 mRNA及Bax mRNA表达的影响,探讨穴位埋线对VD大鼠脑缺血性损伤的神经保护机制。 方法 采用改良Pulsinelli’s 4血管阻断法建立VD大鼠模型,随机数表法分为VD模型组、穴位埋线组、尼莫地平组,并设置假手术组作为对照。两个治疗组分别施行穴位埋线和尼莫地平治疗。Morris 水迷宫检测各组大鼠学习记忆能力后,取含海马的脑组织,TUNEL法检测海马CA1区神经细胞凋亡,原位杂交法检测其Bcl-2 mRNA及Bax mRNA表达,观察并比较各组大鼠海马神经细胞凋亡、蛋白表达的变化。 结果 VD模型组大鼠海马CA1区可见大量凋亡细胞、Bcl-2 mRNA 表达降低、Bax mRNA 的表达增高,与假手术组比较差异均有统计学意义（P＜0.01）。VD模型组表现出明显的学习记忆障碍,与假手术组相比差异有统计学意义（P＜0.01）;与VD模型组比较,穴位埋线组大鼠学习记忆能力显著提高（P＜0.01）,CA1区凋亡细胞明显减少（P＜0.01）,可见一定数量的Bcl-2 mRNA阳性细胞表达,而Bax mRNA阳性细胞表达较少。 结论 穴位埋线可通过上调VD大鼠海马CA1区Bcl-2 mRNA的表达、下调Bax mRNA 的表达,抑制海马神经细胞凋亡,改善VD大鼠学习记忆能力。     

大鼠血管性痴呆模型动物中海马神经元凋亡和蛋白表达的研究

目的:观察不同时点分别结扎左、右颈总动脉建立大鼠血管性痴呆模型中海马CA1区神经元凋亡和Bcl-2及Bax蛋白表达的影响,探讨其在血管性痴呆发病过程中的作用。方法:采取间隔3 d分2次结扎双侧颈总动脉建立血管性痴呆模型,术后4周用TUNEL法检测海马CA1区神经元凋亡,用免疫组织化学法检测其Bcl-2及Bax蛋白表达。结果:模型组大鼠海马CA1区可见大量凋亡神经元;模型组Bcl-2及Bax蛋白表达明显增加,与假手术组比较差异均有显著意义(P<0.05)。结论:此血管性痴呆模型大鼠中海马CA1区神经元大量凋亡丢失,可能是导致血管性痴呆的病理基础。     

丁苯酞通过SIRT1/NF-κB信号通路抑制阿尔茨海默病大鼠海马神经元凋亡的机制研究

目的:探讨丁苯酞通过SIRT1/NF-κB信号通路对阿尔茨海默病(AD)大鼠海马神经元凋亡的影响及其机制。方法:采用氯化铝(AlCl_3)灌胃联合腹腔注射D-半乳糖的方法制备AD大鼠模型,分别给予25 mg/kg(低剂量)、50 mg/kg(中剂量)和100 mg/kg(高剂量)丁苯酞处理后,采用HE染色、流式细胞术和Western blot法分别观察丁苯酞对各组大鼠海马神经元形态、凋亡率及凋亡相关蛋白Bcl-2、Bax和cleaved caspase-3及SIRT1/NF-κB信号通路相关蛋白表达的影响;分别以SIRT1激动剂SRT1720和抑制剂sirtinol处理大鼠后,观察SIRT1/NF-κB信号通路在海马神经元凋亡中的作用;在给予50 mg/kg丁苯酞的基础上,再给予sirtinol作用,观察丁苯酞调控SIRT1/NF-κB信号通路对神经元凋亡的影响。结果:丁苯酞可改善AD大鼠海马神经元的形态,显著抑制AD大鼠海马神经元凋亡及Bax和cleaved caspase-3蛋白的表达,而促进Bcl-2蛋白表达和SIRT1/NF-κB信号通路的激活(P0.05)。SIRT1激动剂SRT1720可显著促进Bcl-2蛋白表达和SIRT1/NF-κB信号通路的激活,并抑制海马神经元凋亡及Bax和cleaved caspase-3蛋白的表达(P0.05);而SIRT1抑制剂sirtinol的作用则与SRT1720相反。经sirtinol处理后,丁苯酞对海马神经元凋亡及Bax和cleaved caspase-3蛋白表达的抑制作用及对Bcl-2蛋白表达的促进作用均显著减弱(P0.05)。结论:丁苯酞可通过激活SIRT1/NF-κB信号通路下调Bax和cleaved caspase-3,并上调Bcl-2蛋白的表达抑制AD大鼠海马神经元凋亡。     

高压氧治疗对癫痫大鼠血清白细胞介素1β、白细胞介素2、白细胞介素8、肿瘤坏死因子α水平及海马神经元Bax及Bcl-2表达的影响

目的：探究高压氧治疗对癫痫大鼠血清白细胞介素（IL）1β、IL-2、IL-8、肿瘤坏死因子α（TNF-α）的表 达及海马神经元细胞凋亡基因表达的影响。方法：采用随机数字法将36 只成年健康SD大鼠进行编号,1 ～ 12 号 实验动物为空白对照组,剩余动物接受匹罗卡品建立癫痫模型,根据Racine 评分作为完成标准,随机选取12 只 接受高压氧治疗,利用ELISA 法检测实验动物血清IL-1β、IL-2、IL-8、TNF-α 表达水平,利用免疫印迹和免疫 荧光染色检测实验动物海马神经元细胞凋亡蛋白Bax 和Bcl-2 表达。结果：与空白对照组相比,癫痫模型组大鼠 血清IL-1β、IL-8、TNF-α 表达水平均显著升高,IL-2 表达水平明显降低;经过高压氧治疗后,与癫痫模型组相比, 实验大鼠血清IL-1β、IL-8、TNF-α 表达水平显著降低,IL-2 表达水平明显升高。免疫印迹和免疫荧光染色结果 显示,与空白对照组相比,癫痫模型组促凋亡蛋白Bax 表达显著升高,而抗凋亡蛋白Bcl-2 表达显著下降,经过 高压氧治疗后,Bax 蛋白水平显著下调,而Bcl-2 蛋白水平显著上调。结论：高压氧治疗可减少癫痫大鼠血清IL- 1β、IL-8、TNF-α 的表达,增加IL-2 的表达, Bax 蛋白水平显著下调而Bcl-2 蛋白水平显著上调,因此本研究可作 为高压氧治疗癫痫病作用机制之一。     

母体肢体缺血预处理对宫内窘迫胎鼠复氧后海马神经元凋亡的影响*

 目的： 观察母体的肢体缺血预处理(LIP)对宫内窘迫胎鼠复氧后海马神经元凋亡的影响。方法: 采用微动脉夹阻断母鼠通向子宫和卵巢的动静脉15 min后开放以制备胎鼠宫内窘迫模型。孕19 d SD大鼠12只,随机分为4组：空白对照（S）组、LIP组、胎儿窘迫（FD）组和LIP+FD组。各组母鼠再灌注2 d时剖宫取活胎鼠12只断头取脑。TUNEL法测定胎鼠海马CA1区神经元凋亡情况,计算细胞凋亡指数;免疫组化法和Western blotting法测定Bcl-2和Bax蛋白的表达。结果: 与S组比较,FD组和LIP+FD组胎鼠海马CA1区神经元凋亡指数增加（P＜0.05）,Bcl-2蛋白表达增加,Bax蛋白表达增加,Bcl-2/Bax比值降低;与S组比较,LIP组胎鼠海马CA1的 Bcl-2、Bax蛋白表达及Bcl-2/Bax蛋白表达的比值无明显改变（P＞0.05）;与FD组比较,LIP+FD组细胞凋亡指数降低（P＜0.05）,Bcl-2/Bax比值增加。结论: 母体肢体缺血预处理减轻了宫内窘迫胎鼠复氧后海马神经元的凋亡,其机制可能与Bcl-2蛋白表达的上调相关。     

丁苯酞减轻阿尔茨海默病大鼠海马神经元凋亡

目的探讨丁苯酞对阿尔茨海默病(AD)大鼠海马神经元凋亡的影响。方法将大鼠分为对照组、AD模型组(腹腔注射D-半乳糖和三氯化铝)、丁苯酞低、中和高剂量组(25、50和100 mg/kg)为灌胃干预组,每组12只。流式细胞仪检测海马神经元凋亡,HE观察海马CA1区神经元形态变化; real-time-PCR检测海马神经元中Tau蛋白及凋亡因子Bcl-2、bax和caspase-3 mRNA表达; Western blot检测海马神经元中mTOR、AKT和GSK-3β蛋白表达。结果与对照组比较,模型组大鼠第1～5天逃避潜伏期及第1次找到原始平台时间明显延长,跨越原始平台次数明显减少,海马神经元凋亡率均明显升高,模型组Tau、Bax、caspase-3 mRNA表达升高; Bcl-2 mRNA表达及mTOR、AKT、GSK-3β蛋白表达量均降低(P0. 05);与模型组比较,丁苯酞低、中和高剂量组第3～5天逃避潜伏期及第一次找到原始平台时间明显缩短,跨越原始平台次数明显增加,海马神经元凋亡率均明显降低,Tau、Bax、caspase-3 mRNA降低,Bcl-2 mRNA及mTOR、AKT、GSK-3β蛋白表达量均升高(P0. 05)。结论丁苯酞具有显著的抑制AD大鼠海马神经元凋亡作用,其作用机制可能与抑制Tau蛋白、调节凋亡因子和激活mTOR/AKT/GSK-3β信号通路有关,但具体机制还有待进一步分析。     

远志对糖尿病大鼠海马神经细胞凋亡相关蛋白表达的影响

目的:探讨远志对糖尿病(DM)大鼠海马神经细胞的影响.方法:大鼠随机分为正常对照组、糖尿病模型组、远志治疗组和远志预防组.除正常对照组外均建立链脲佐菌素致糖尿病模型.远志预防组大鼠在建模同时给予远志灌胃6周;此后给予远志治疗组大鼠远志灌胃6周.采用尼氏染色法观察大鼠海马CA1区神经细胞形态并计数神经细胞数量;采用免疫印迹检测大鼠海马组织Bcl-2和Bax的表达.结果:与正常对照组比较,糖尿病模型组大鼠海马CA1区神经细胞形态异常,数量减少,Bcl-2表达降低,Bax表达升高;与糖尿病模型组比较,远志治疗组与远志预防组大鼠海马CA1区生存神经细胞数量增多,Bcl-2表达升高,Bax表达降低.结论:远志可通过上调糖尿病大鼠海马Bcl-2的表达,并减少Bax的表达,抑制海马神经细胞凋亡,促进其存活,从而发挥对糖尿病大鼠海马损伤的预防保护作用.     

3-甲基腺嘌呤对Aβ(25-35)引起的海马神经元凋亡的影响

目的: 探讨阿尔茨海默病(AD)大鼠海马神经元自噬对凋亡的影响。方法: SD大鼠随机分成模型组、自噬抑制剂3-甲基腺嘌呤(3-MA)预处理组和对照组。模型组大鼠用立体定位技术对海马CA1区微量注射Aβ(25-35)造成AD模型;3-MA预处理组大鼠在注射Aβ(25-35)之前对海马CA1区微量注射3-MA。Morris水迷宫检测大鼠记忆水平;行为学测试后,观察海马神经元超微结构变化、自噬泡的形成、beclin-1的表达以及细胞凋亡情况。结果: 3-MA预处理组与模型组比较,大鼠学习记忆能力显著下降(P<0.05),海马神经元凋亡率显著增加(P<0.05),而beclin-1的表达量减少;模型组海马神经元可见双层膜包裹形成的自噬泡,神经元破坏程度明显轻于3-MA预处理组。结论: 抑制神经元自噬水平增加神经元凋亡;诱导神经元自噬可能是防治阿尔茨海默病的一个潜在方法。     

颈动脉狭窄影响大鼠认知功能及海马神经元凋亡

目的:研究改善颈动脉狭窄对大鼠认知功能、海马神经元凋亡及bcl-2、Bax蛋白表达的影响.方法:采用SD大鼠制作颈动脉狭窄模型,2周后将颈动脉狭窄解除,4周后各组采用Morris水迷宫检测记忆能力、HE染色观察神经元形态学变化、TUNEL法观察海马神经元凋亡、免疫组织化学检测Bcl-2、Bax蛋白表达.结果:对照组与假手术组比较,大鼠记忆能力明显下降(P<0.01),神经细胞凋亡率明显增高(P<0.01),Bcl-2和Bax免疫阳性细胞数增多(P<0.01),改善狭窄组较对照组记忆能力明显改善(P<0.01),神经细胞凋亡率明显低于对照组(P<0.05),Bcl-2免疫阳性细胞数明显增多(P<0.05),Bax免疫阳性细胞数明显减少(P<0.05).结论:改善颈动脉狭窄可提高大鼠海马Bcl-2蛋白的表达,抑制海马神经细胞凋亡,改善认知功能障碍.     

Cure and care of neurosis. I. Cure.

Behavioural psychotherapy has long historical roots. Recently it has led to effective treatment for selected neuroses, including phobic, obsessive-compulsive and sexual disorders. Potent therapy has become a tool of experimental psychopathology which advances theory and practice. A pervasive principle is exposure of the patient to those stimuli which evoke his discomfort until this subsides. Level of arousal during exposure does not affect outcome. Theoretical issues are reviewed which decide when exposure will be sensitizing or habituating. Both psychoanalytical and conditioning models of neurosis are out of date, and models derived more directly from clinical experiment are becoming possible. The aetiology of phobias and rituals can be seen as failed extinction rather than enhanced acquisition. Relevant phylogenetic and biological factors are discussed. At the other extreme, well-documented faith-healing indicates huge gaps in our knowledge of psychotherapy.     

Immunogenicity of surfactant. II. Porcine and bovine surfactants.

Protein-containing surfactants of human and animal origin are being used increasingly to treat neonatal and adult respiratory distress syndromes. This trend led us to examine the antigenicity of two important preparations of animal surfactant, cow lung surfactant extract (CLSE) and a porcine surfactant preparation, Curosurf. We describe here 15 monoclonal antibodies against Curosurf and four against CLSE. Antibodies were studied by Western blot analysis to determine their ability to recognize protein components of their respective surfactant preparations. They were also tested for their ability to inactivate surfactant in vitro, assayed using the pulsating bubble surfactometer. Several antibodies directed against CLSE or Curosurf functionally inactivate the surfactant to which they were raised. We determined the degree of immunologic cross-reactivity between antibodies directed to CLSE and Curosurf against the other surfactant and also against human surfactant, both by Western blot and by examining functional inactivation in vitro. Antibodies to these animal surfactants that are commonly used therapeutically may inactivate the specific animal surfactant to which they were raised, as well as human and other surfactants. Generally, when antibodies inactivate surfactant from more than one animal species, they inactivate heterologous surfactants comparably to the extent to which they inactivate the surfactant to which they are directed. Immune complexes between anti-surfactant antibodies and surfactant have been described in the course of neonatal respiratory distress syndrome. The potential pathophysiological importance of anti-surfactant antibodies may therefore lie in their ability to inactivate administered surfactant, other similar surfactants and endogenous surfactant. In so doing, these antibodies may potentiate surfactant deficiency or pulmonary injury initiated by other stimuli.     

A review of biointeractions of Ni and Mg. II. Immune system and oncology.

Reports of interactions, in vivo and in vitro, between Ni and Mg in humoral and cellular immunity, hypersensitivity and inflammation, and in tumourigenesis are explored from a mechanistic viewpoint. Although Mg is present in much larger concentrations in normal mammalian systems than Ni, similar chemical and physical properties may allow Ni to exchange for Mg at reactive sites with damaging consequences to living organisms. Consequences of such exchanges could involve reduced immunocompetence and related carcinogenic transformation of cells. Mg status and environmental exposure to Ni are conceivable antecedents to possible biological sequelae in humans.     

Gel casting of resorbable polymers. 1. Processing and applications.

A gel casting technique based on resorbable, synthetic, alpha-polyesters (or lactide-glycolide polymers) is described for producing medical implants such as bone graft substitutes and timed-release carriers for medication. This solution-based method enables production of thick-section solid and microporous materials, and blending of polymers and particulate fillers. Implant degradation rate, for example, may be adjusted by variation of polymer type, molecular weight range, crystallinity and morphology. Gel casting conditions are reported for solid and microporous materials and processing characteristics are interpreted in terms of established crystallization and dissolution behaviour of polymers.     

Freeze-fracture ultrastructure of rat C.N.S. and P.N.S. nonmyelinated axolemma

Summary The axolemma of nonmyelinated fibres from the corpus callosum and cerebellar cortex (C.N.S.) and the vagus nerve (P.N.S.) was investigated with freeze-fracture electron microscopy. The major observations of this study are as follows: (1) there is a highly asymmetrical distribution of intramembranous particles between the E- and P-fracture faces in both C.N.S. and P.N.S. fibres; (2) the total number of particles on the P-faces of all axonal types studied is considerably greater than that on the E-face; (3) the number of particles on the E-faces of C.N.S. axons is greater than that on the E-faces of P.N.S. axons; and (4) the percentage of large (>9.6 nm) particles is greater on the E-face than on the P-face regardless of the axon studied. The results are compared with previous freeze-fracture investigations on the nodal and internodal membranes of myelinated fibres.