β-蜕皮甾酮对APP/PS-1转基因小鼠学习记忆能力及海马Bcl-2、Bax蛋白表达的影响

目的观察β-蜕皮甾酮对APP/PS-1转基因小鼠学习记忆能力及海马Bcl-2、Bax蛋白表达的影响。方法取C57小鼠6只及APP/PS-1转基因小鼠12只,C57小鼠为对照组(Control)、APP/PS-1小鼠12只随机分为模型组(APP/PS-1)和实验组(APP/PS-1+20HE),实验组APP/PS-1转基因小鼠按14.4mg/kg的剂量灌胃β-蜕皮甾酮,每日一次,对照组及模型组小鼠灌胃等量蒸馏水,连续灌胃8周,Morris水迷宫检测小鼠学习记忆能力,尼氏染色观察各组小鼠海马组织病理学变化,免疫组化法观察海马Bcl-2和Bax蛋白表达的变化。结果模型组小鼠与对照组小鼠相比,认知能力明显下降(P<0.01),海马Bcl-2和Bax蛋白表达均增加。实验组与模型组相比,学习记忆能力明显改善(P<0.01),Bcl-2蛋白的表达增加,Bax蛋白的表达减少。结论β-蜕皮甾酮能够改善转基因小鼠的学习记忆能力,可能与促进海马Bcl-2蛋白表达和抑制Bax蛋白表达有关。     

锌螯合剂对匹罗卡品致痫小鼠行为学及海马神经元的影响

目的:研究锌螯合剂氯碘羟喹对匹罗卡品致痫小鼠行为学及海马神经元的影响。方法:将CD1小鼠随机分成3组,对照组小鼠腹腔注射等量的生理盐水;癫痫组小鼠腹腔注射匹罗卡品300 mg/kg;氯碘羟喹治疗组(治疗组)小鼠腹腔注射匹罗卡品300 mg/kg,30 min之后腹腔注射氯碘羟喹15 mg/kg。观察各组小鼠的癫痫发作次数。Morris水迷宫实验测定逃避潜伏期和目标象限停滞时间。应用尼氏染色计数海马神经元数量;利用免疫组织化学方法、免疫印迹结合图像分析技术检测凋亡基因caspase-3在各组小鼠海马的表达变化。结果:与对照组相比,其他2组小鼠的癫痫发作次数明显增多,发作级别明显增高;其中,治疗组小鼠的发作次数和发作级别均低于癫痫组。水迷宫实验结果显示,与对照组相比,其他2组小鼠的逃避潜伏期明显延长,目标象限停滞时间明显缩短;其中,治疗组的逃避潜伏期比癫痫组短,目标象限停滞时间比癫痫组长。其他2组小鼠海马神经元数量明显少于对照组;治疗组的海马神经元数量比癫痫组多。其他2组小鼠海马caspase-3表达明显高于对照组,阳性细胞数量增多,光密度增加;治疗组海马caspase-3的表达低于癫痫组,阳性细胞数量减少,光密度降低。结论:锌螯合剂氯碘羟喹能抑制癫痫发作,保护海马神经元,提高癫痫小鼠的学习记忆能力。     

铝负荷对小鼠脑内ChAT/AchE的影响及蜕皮甾酮干预作用

目的 :观察铝负荷对小鼠脑内胆碱乙酰化酶(ChAT)和胆碱酯酶 (AchE)的影响及铝负荷对小鼠被动回避实验、空间辨别实验的影响以及蜕皮甾酮(Ecdysterone,ECR)的干预作用。方法 :NIH小鼠(♂ ) 90只随机分为对照组 (control) ,模型组 (Aβ) ,治疗组 [ECR 4mg·kg (Aβ +ECR4) ,ECR 8mg·kg(Aβ +ECR8) ,Nimodipine 7.2mg·kg(Aβ+Ni) ],侧脑室内微注射铝盐 3 μL(1 5 0 μmol·L 1,连续 5天 ,1次 /天 ) ,免疫组化SP法和化学法分别检测皮层及海马内的ChAT表达情况及AchE活性。被动回避实验用跳台法 ,空间辨别实验用水迷宫法。结果…     

β-蜕皮甾酮促进小鼠前成骨细胞体外增殖及诱导成骨分化

文题释义：β-蜕皮甾酮：又称蜕皮激素、20-羟基蜕皮甾酮,分子式C₂₇H₄₄O₇,为黄棕色至白色粉末,味苦;主要存在于昆虫、蚕、露水草、牛膝、川牛膝等动植物体内,具有调节血糖血脂、促进胶原蛋白合成、抗疲劳、促进细胞生长和刺激真皮细胞分裂等作用,目前广泛应用于化妆品、医疗以及养殖业等领域。骨形态发生蛋白质2：是从脊椎动物骨骼基质中分离提纯的蛋白质,具有内肽酶活性、表皮生长因子模体,同源二聚体之间以二硫键相连,属于转化生长因子β家族,能诱导骨与软骨形成。骨桥蛋白：存在于矿化和活性沉积区,是一种含有Arg-Gly-Asp(RGD)结构的酸性糖蛋白,它与诱导成骨细胞成熟表型表达和矿化骨基质形成的活性蛋白密切相关。在成矿阶段,骨连接素、纤连蛋白与骨桥蛋白的表达高度相关。Ⅰ型胶原是钙盐沉积和细胞附着的支架,可促进细胞附着并刺激细胞分化。背景：β-蜕皮甾酮作为“植物类雌激素”不仅具有刺激蛋白质合成,促进碳水化合物和脂质代谢,缓解高血糖、高脂血症,以及保护内皮细胞免于凋亡并诱导其增殖的多种生物活性,而且有学者报道其在治疗骨质疏松症、骨折和其他骨骼炎症性疾病方面也有着重要的作用。 目的：观察β-蜕皮甾酮对小鼠前成骨细胞(MC3T3-E1细胞)体外增殖的影响,以及在安全剂量下,β-蜕皮甾酮是否对该细胞具有诱导成骨分化的作用。方法：取第4代MC3T3-E1细胞在成骨诱导分化培养基中培养7,10,14,21,28 d后,检测细胞不同时间段成骨分化蛋白(碱性磷酸酶、Ⅰ型胶原蛋白、骨桥蛋白以及钙化结节)的表达量,以鉴定该细胞是否具有成骨分化的能力;然后将MC3T3-E1细胞接种于含不同终浓度β-蜕皮甾酮(0.01,0.1,1,10,100 µmol/L)的诱导培养基中,分别于第1,2,3,4,5,6,7天利用CCK8法检测细胞的增殖活性;最后设置对照组(普通诱导培养基组)和实验组(普通诱导培养基+β-蜕皮甾酮),在相同条件下进行培养,并测定不同时间段各组细胞成骨标志蛋白的表达量。结果与结论：①MC3T3-E1细胞在成骨诱导培养基刺激下,第10天碱性磷酸酶染色以及Ⅰ型胶原蛋白荧光染色表达较高,同时碱性磷酸酶活性检测也验证了这一结果(P < 0.05);诱导培养第14天骨桥蛋白免疫细胞化学染色也有明显表达;茜红素染色显示成骨诱导后的细胞较对照组结节数量明显增加,第28天比第21天的钙结节形成数目更多、直径更大、颜色更深;②CCK 8法测得β-蜕皮甾酮对MC3T3-E1细胞作用5 d后增殖活性达到最佳,促增殖活性最佳剂量为0.01 µ mol/L、0.1 µmol/L,2种浓度之间差异无显著性意义(P > 0.05);③实验组细胞经β-蜕皮甾酮诱导培养第10天较对照组细胞碱性磷酸酶、Ⅰ型胶原蛋白表达更高;第14天实验组细胞内骨桥蛋白、骨钙素表达更高;第28天2组钙结节染色无明显差异;④结果说明,β-蜕皮甾酮能促进MC3T3-E1细胞体外增殖,且在安全剂量下能提高MC3T3-E1细胞向成骨细胞分化的能力。ORCID: 0000-0001-5641-6353(严才平) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

蜕皮甾酮对肿瘤坏死因子致人脐静脉内皮细胞损伤的影响

目的和方法：实验以培养的人脐静脉内皮细胞（ＨＵＶＥＣ）为研究对象，应用大剂量肿瘤坏死因子（ＴＮＦ）造成ＨＵＶＥＣ损伤，观察培养上清液血管紧张素转换酶（ＡＣＥ）及ＨＵＶＥＣ的超微结构变化，研究蜕皮甾酮（ＥＤＳ）对内皮细胞损伤的影响。结果：大剂量ＴＮＦ作用于ＨＵＶＥＣ时，其培养上清液的ＡＣＥ非常明显增高，超微结构破坏；应用ＥＤＳ，则培养上清液的ＡＣＥ有所减少、超微结构的损害程度有所减轻。结论：ＥＤＳ能够减轻人血管内皮细胞的损伤，揭示了ＥＤＳ在抗血管内皮细胞损伤方面的价值。     

低频经颅磁刺激对锂-匹罗卡品致痫大鼠痫性发作的影响

目的：探讨低频经颅磁刺激（TMS）对癫痫大鼠痫性发作的影响。方法：对50只大鼠进行TMS,根据不同刺激频率将大鼠随机分为5个组（每组10只）：0Hz组（假刺激组）、0．3Hz组、0．5Hz组、0．8Hz组、1．0Hz组,分组进行低频重复TMS预处理后,制作氯化锂一匹罗卡品癫痫模型,观察各组造模后70min内痼性发作的潜伏期和痫性发作行为表现。结果：0．3Hz组、0．5Hz组、0．8Hz组及1．0Hz组的痫性发作潜伏期[分别为（16．30±3．10）min、（18．60±3．50）min、（16．40±3．60）min和（15．90±4．10）mini,均比0Hz组[（12．80±3．20）mini延长,差异均有统计学意义（P〈0．05或P〈0．01）,其中0．5Hz组最长;0．5Hz组大鼠出现的痫性行为表现为点头和（或）甩尾、须动和（或）头面抽动、前肢和（或）后肢抽动、全身抽动、窜动的次数均比0．3Hz组、0．8Hz组、1．0Hz组少,整体发作程度轻,差异有统计学意义（P〈0．05）。结论：适量的低频的TMS可延缓癫痫发作潜伏期、减少癫痫发作次数以及减轻癫痫发作程度,具有抗痢作用。     

匹罗卡品致痫大鼠脑内NF-κB及COX-2的表达

为探讨癫痫大鼠脑内核转录因子NF-κB及炎性因子环氧化酶-2(COX-2)的表达,本研究采用匹罗卡品(30mg/kg)诱导大鼠癫痫发作后,通过免疫组织化学和Westernblot方法,对癫痫大鼠脑内,主要是海马NF-κB和COX-2的表达进行了观察。结果显示:匹罗卡品诱导大鼠癫痫发作后,大鼠脑内尤其是海马神经元的胞浆和胞核内NF-κBp65免疫反应阳性增强,核移位增多,蛋白产物表达增多;COX-2免疫反应阳性明显增强,蛋白产物表达增多。以上结果提示,癫痫发作后可引起核转录因子NF-κB的活化及炎性因子COX-2的高表达。     

内质网应激介导蜕皮甾酮对H2 O2 诱导的心肌细胞损伤的保护作用

目的:探讨蜕皮甾酮(EDS)如何通过调节内质网应激对过氧化氢(H2 O2 )诱导的心肌细胞损伤起保护作用。方法:大鼠H9c2 心肌细胞随机分为对照(Control)组,正常心肌细胞;H2 O2 组,不同浓度H2 O2(1*10-3 、1*10-4 、1*10-5 mol/ L)诱导损伤细胞;EDS 组,在H2 O2 组基础上,加入不同浓度的EDS(25、50、100 μmol/ L)处理。MTT 法和流式细胞术分别检测实验各组细胞活力及细胞凋亡率。Western blot 检测各组细胞中Bcl-2、Bax、cleaved-caspase-3、caspase-4、caspase-7、caspase-12、 GRP78 和CHOP 的蛋白水平,同时检测各组细胞中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。结果:与Control 组相比,H2 O2 组的细胞活力减弱,凋亡率升高,MOD 含量升高,SOD 活性降低,GRP78 和CHOP 的表达升高(均P<0.05)。H2 O2 组加入EDS 处理后,细胞活力提升,凋亡率下降,MOD 含量降低,SOD 活性升高,GRP78 和CHOP 的表达降低(均P<0.05)。结论:通过抑制内质网的应激过程,蜕皮甾酮能减轻H2 O2 诱导的心肌细胞损伤。     

氯化锂匹罗卡品诱导癫痫大鼠海马神经元GPER1表达的变化

目的:观察G蛋白偶联雌激素受体1(GPER1)在癫痫大鼠海马神经元中表达的变化。方法:成年雄性SD大鼠分成对照组(control)和癫痫组(epilepsy),利用腹腔注射氯化锂-匹罗卡品方法制备癫痫模型,分别在1、2、3、7、14 d和28 d,利用Morris水迷宫检测大鼠学习记忆能力,利用尼氏染色观察大鼠海马神经元形态变化;利用免疫组化和Western Blot技术观察GPER1在海马的表达。结果:水迷宫结果显示,与Control组相比,造模14 d的大鼠逃逸潜伏时间明显延长(P 0. 05),穿越目标象限区域的次数较Control组显著降低(P 0. 05)。尼氏染色结果显示:与Control组相比,造模1 d和2 d的大鼠CA1及CA3区锥体细胞层细胞及DG区颗粒细胞层细胞体积缩小,细胞间距增加,尼氏染色减弱;造模3和7 d的大鼠细胞体积明显缩小,细胞间隙明显增大,尼氏染色加深,CA1及CA3细胞数量明显减少;造模14 d和28 d的大鼠神经元体积逐渐向正常恢复,但仍较Control组小。免疫组化结果显示:GPER1免疫阳性细胞以海马锥体细胞和齿状回颗粒细胞为主,主要分布在细胞膜。与Control组相比,造模2 d和3 d的大鼠海马CA1及CA3区GPER1表达增加(P 0. 05),7 d后增加最明显(P 0. 01),14、28 d后表达下降;在DG区,造模3 d及7 d的大鼠GPER1表达增加(P 0. 05),14 d后表达下降(P 0. 05),28 d大鼠无显著差异。Western Blot结果显示:与Control组比较,造模2 d和3 d的大鼠GPER1相对表达量开始增高,7 d后明显增高(P 0. 05),14 d及28 d的大鼠表达降低。结论:GPER1在海马神经元的表达随着神经元损伤的加重而增高,随着神经元损伤的恢复逐渐降低,提示其表达变化与神经元的损伤与修复有关。     

氯碘羟喹抑制癫痫小鼠海马神经元凋亡的实验研究

目的研究氯碘羟喹对癫痫持续状态模型小鼠海马神经元凋亡的影响。方法 18只CD1小鼠随机分为3组:对照组(Con)给予腹腔注射生理盐水;癫痫持续状态组(SE)给予腹腔注射匹罗卡品(300mg/kg);氯碘羟喹治疗组(Cli)在腹腔注射匹罗卡品(300mg/kg)后30min给予腹腔注射氯碘羟喹(15mg/kg·d)。每天观察动物的行为学变化,于7d后取海马组织。应用尼氏染色和TUNEL染色检测氯碘羟喹对癫痫持续状态小鼠海马神经元凋亡的影响。结果癫痫持续状态组动物萎靡,自发性痫性发作频繁;氯碘羟喹治疗组动物萎靡状态明显改善,自发性痫性发作次数明显减少。尼氏染色和TUNEL染色发现,与癫痫持续状态组相比,氯碘羟喹治疗组能改善海马神经元的细胞损伤,降低凋亡阳性细胞百分比(<0.01)。结论氯碘羟喹能抑制癫痫持续状态小鼠海马神经元凋亡。     

阿魏酸钠对SNP诱导的凋亡海马神经元bcl-2、bax基因表达的影响

目的：探讨阿魏酸钠(SF)对一氧化氮（NO）供体硝普钠(SNP)引起的大鼠海马神经元凋亡及bcl-2、bax基因表达的影响。方法:采用SD大鼠海马神经元原代培养，经终浓度分别为10、20、40、80、120、160 μmol/L SF预处理后，用50 μmol/L SNP处理24 h，采用MTT法检测细胞存活率，Hoechst 33258荧光染色及DNA琼脂糖凝胶电泳分析等方法检测凋亡，Western blotting及RT-PCR检测bcl-2、bax基因表达。结果:不同剂量SF (10-160 μmol/L)预处理6 h可显著提高神经元的存活率，减少SNP引起的核固缩、凝聚和碎裂现象；DNA凝胶电泳图谱未见典型的“梯状”改变；增加bcl-2 mRNA及蛋白的表达, 降低bax mRNA及蛋白的表达。结论:SF 抑制NO供体SNP诱导的海马神经元凋亡，其机制可能与其增加Bcl-2蛋白表达，降低Bax蛋白表达，增高Bcl-2/Bax的比值有关。     

Interrelationship between Epstein-Barr virus infection in gastric carcinomas and the expression of apoptosis-associated proteins

AIMS: Epstein-Barr virus (EBV) and its associated proteins may be protective against the occurrence of apoptosis that would normally inhibit cancer development and progression. Alternatively, the viral infection may cause altered or mutated expression of oncogenes or tumour suppressor genes that are necessary for tumour development, an action that may also involve apoptosis. In this study, a relationship was sought between occurrence of EBV infection, expression of apoptosis-associated proteins (tumour suppressor gene p53 and oncogenes c-myc and bcl-2) and levels of cell death (apoptosis or necrosis) in 119 cases of gastric carcinoma. METHODS AND RESULTS: The EBV status of the gastric carcinomas (using the EBV-encoded small RNA I (EBER-1) and in-situ hybridization), stage and grade of tumour and sex of patients were compared for bcl-2, p53 and c-myc expression patterns. EBER-1 was detected in approximately 20% of cases studied. There was no significant correlation between levels of cell death in the tumour tissue and EBV status. In the protein analyses, development and progression of gastric carcinoma, with or without EBV infection, was independent of bcl-2 expression. However, in gastric cancers with EBV infection, p53 overexpression was inhibited and c-myc expression was increased in early stage cancers, in comparison with decreased c-myc expression in late stage cancers. CONCLUSIONS: The p53 and c-myc expression patterns indicate that EBV-infected gastric carcinomas are less likely to have a natural regression via apoptosis at an early stage and explain, in part, the resistance to treatment of late stage of gastric cancers.     

儿童急性白血病骨髓活检组织中bcl-2、CD20、CD79a的表达及临床意义

目的 探讨儿童急性白血病骨髓活检组织中bcl-2、CD20、CD79a的表达与发病和预后的关系.方法 对31例儿童急性白血病及10例对照组病例骨髓活检组织进行bcl-2、CD20、CD79a免疫组化检测.结果 bcl-2、CD20、CD79在初发白血病骨髓活检组织中的表达均增高,与对照组差异存在统计学意义(P＜0.05).缓解组骨髓活检组织6例bcl-2阳性表达中2例临床复发(33.3%),4例临床缓解(66.7%),13例bcl-2阴性表达临床均CR(P＜0.05).CD20、CD79a阳性表达与临床复发/CR差异无显著性意义(P＞0.05).结论 儿童急性白血病骨髓活检组织中bcl-2的检测可作为协助临床疗效判断、指导治疗的参考依据.     

Deletion of puma protects hippocampal neurons in a model of severe status epilepticus

Prolonged seizures (status epilepticus) can activate apoptosis-associated signaling pathways. The extent to which such pathways contribute to cell death might depend on the insult intensity, whereby the programmed or apoptotic cell death component is reduced when seizures are more severe or protracted. We recently showed that mice lacking the pro-apoptotic Bcl-2 homology domain 3-only protein Puma (Bbc3) were potently protected against damage caused by status epilepticus. In the present study we examined whether Puma deficiency was protective when the seizure episode was more severe. Intra-amygdala microinjection of 1 μg kainic acid (KA) into C57BL/6 mice triggered status epilepticus that lasted about twice as long as with 0.3 μg KA prior to lorazepam termination. Hippocampal damage was also significantly greater in the higher-dose group. Over 80% of degenerating neurons after seizures were positive for DNA fragmentation assessed by terminal deoxynucleotidyl dUTP nick end labeling (TUNEL). Microscopic analysis of neuronal nuclear morphology in TUNEL-positive cells revealed the proportion displaying large rounded clumps of condensed chromatin was ∼50% lower in the high-dose versus low-dose KA group. Nevertheless, compared to heterozygous and wild-type mice subject to status epilepticus by high-dose KA, neuronal death was reduced by ∼50% in the hippocampus of Puma-deficient mice. These data suggest aspects of the apoptotic component of seizure-induced neuronal death are insult duration- or severity-dependent. Moreover, they provide further genetic evidence that seizure-induced neuronal death is preventable by targeting so-called apoptosis-associated signaling pathways and Puma loss likely disrupts caspase-independent or non-apoptotic seizure-induced neuronal death.     

蝎毒抗帕金森病小鼠多巴胺能神经元凋亡

研究蝎毒对帕金森病 (PD)小鼠脑内多巴胺能神经元的抗凋亡作用及其机制。用C5 7BL/ 6 品系小鼠 ,每日于颈部皮下注入 1 甲基 4 苯基 1,2 ,3,6 四氢吡啶 (MPTP ,2 0mg/kg)复制帕金森病模型 ,随机分为模型组及蝎毒 (SV)提取液高 (2 0 0mg/kg)、低剂量 (2 0mg/kg)治疗组 ;另设对照组 (NS)和空白给SV高、低剂量组。各组均为 8只。采用爬杆、游泳行为学检测小鼠的运动功能障碍 ;应用DNA断裂点标记法 (TUNEL)检测黑质致密部 (SNc)多巴胺 (DA)能神经元的凋亡 ;利用免疫细胞化学法检测Bcl 2 /Bax基因的表达。结果表明 :SV能改善PD小鼠运动功能障碍 ;模型组SNc部可见大量TUNEL阳性细胞 (P <0 0 1) ,治疗组TUNEL阳性细胞数均明显减少 (P <0 0 1) ;模型组SNcBcl 2免疫反应活性 (IR)阳性神经元数明显减少 (P <0 0 1) ,Bax IR阳性神经元数明显增多 (P <0 0 1) ,治疗药组未见Bcl 2 IR阳性神经元数明显减少或Bax IR阳性神经元数明显增多。提示 :SV可能通过上调Bcl 2、下调Bax基因表达抑制DA能神经元凋亡。     

海人酸致癫痫大鼠海马结构中noggin表达变化

目的:研究海人酸(kainic acid,KA)侧脑室注射并诱发癫痫持续状态(status epilepticus,SE)后大鼠海马结构中noggin基因在大鼠海马的表达变化。方法:大鼠在侧脑室注射KA后1、3、7、14、30和60d等不同时间,采用RT-PCR研究noggin mRNA含量变化,采用免疫组化观察noggin蛋白表达变化。结果:在正常大鼠海马结构中noggin mRNA有少量表达。noggin阳性细胞主要位于齿状回及CA3、CA1区,数量较少。侧脑室注射KA诱发SE后,noggin表达持续升高,3d达到高峰;7d在脑内的表达开始降低;注射后2个月,noggin表达降至术前水平,仅见散在的阳性细胞。结论:侧脑室注射KA并诱发SE后,大鼠海马结构中noggin表达明显增加。     

Prognostic value of apoptosis and apoptosis-associated proteins in salivary gland adenoid cystic carcinoma

The purpose of the present study was to investigate the level of apoptosis and expressions of p53, mdm2 and bcl-2 proteins in salivary gland adenoid cystic carcinoma (ACC) to determine potential relationships among apoptosis, apoptosis-associated proteins and clinical cumulative survival. Thirty-nine formalin-fixed, paraffin-embedded cases, cribriform (17), tubular (13) and solid (9), were studied by immunohistochemistry. Apoptosis detection and analysis were determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL). There was an inverse significance between the apoptotic index (AI) and bcl-2 expression (P = 0.018), whereas no correlation was found between the AI and either p53 or mdm2 expression (P = 0.416 and P = 0.456). Co-expression of p53 and mdm2 was found in 22 cases (P = 0.037). Patients with p53-positive tumors had a worse prognosis than those with p53-negative tumors (P = 0.014). Patients with a high AI had a better cumulative survival than patients with a low AI (P = 0.038). The present study suggests that p53 expression and AI can be useful as prognostic values; bcl-2 protein plays a role in the down-regulation of apoptosis and is also potentially useful as a prognostic parameter in salivary gland ACC.     

D-半乳糖导致小鼠海马基因表达谱变化

本实验采用cDNA芯片研究D-半乳糖皮下注射小鼠海马的基因表达变化。小鼠皮下注射D-半乳糖[50m∥(kg．d)]60d造模，Morris水迷宫测试小鼠行为，分别提取模型组和对照组海马组织总RNA，掺入荧光分子Cy3和Cy5，经逆转录合成cDNA荧光探针与4000点小鼠cDNA基因芯片杂交。结果显示，在4000条待研究的基因中，两组小鼠海马间存在2倍表达差异的有76条，其中上调26条，下调50条。经分析，模型组基因表达谱与老年性痴呆(AD)相关基因表达谱在氧应激、能量代谢相关基因等方面有相似之处。提示D-半乳糖小鼠模型有可能作为拟AD病理模型。     

氨茶碱对哮喘豚鼠嗜酸细胞凋亡及bcl—2 mRNA表达的影响

目的 观察氨茶碱对体外不同密度嗜酸细胞（Eos）凋亡及bcl-2 mRNA表达的影响,探讨其促进哮喘Eos凋亡的机制。方法 卵蛋白激发哮喘豚鼠动物模型48h后行支气管肺泡灌洗,分离不同密度Eos。氨茶碱（AM）（10^-6-10^-3mol/L)干预24h,原位杂交检测Eos的bcl-2 mRNA表达,TUNL法检测细胞凋亡。结果 AM干预24h,可见Eos凋亡增加,以低密度Eos(HEos)为明显,bcl-2 mRNA表达,呈剂量依赖性。bcl-2 mRNA的表达与Eos凋亡呈负相关。结论 AM可抑制bcl-2 mRNA表达,促进Eos凋亡。bcl-2参与了AM对Eos凋亡的调节。     

The extent of apoptosis is inversely associated with bcl-2 expression in premalignant and malignant breast lesions

 

Aims:

In this study we investigated the extent of apoptosis in benign, premalignant and malignant breast lesions and its association with the immunohistochemical expression of bcl-2 oncoprotein.  

Methods and results:

In order to detect apoptotic cells and bodies in tissue sections, the 3'-end DNA labelling method was used. Immunohistochemical staining was performed by using the avidin–biotin–peroxidase complex technique. A monoclonal antibody agains bcl-2 oncoprotein was used and the specificity of the antibody was confirmed by immunoblot analysis. According to the results the extent of apoptosis, as determined by the apoptotic index, was lowest in benign ductal hyperplasias and sclerosing adenoses (0.15% and 0.07%, respectively). It was moderately elevated in atypical hyperplasias and in-situ carcinomas (0.20% and 0.40%, respectively) and highest in invasive carcinomas (0.76%). In ductal invasive carcinomas, grade I lesions showed a lower apoptotic index (0.52%) than grade II (0.72%) and grade III (1.17%) carcinomas. The apoptotic index was not significantly lower in lobular (0.82%) than in ductal invasive carcinomas (0.85%). bcl-2 immunohistochemistry was inversely related to the apoptotic index. In all cases studied the inverse association was very strong ( P  = 0.0004) but it was also present when only carcinomas were analysed ( P  = 0.01). In benign and atypical hyperplasias, bcl-2 positivity was observed in all cases, but such cases were less frequent in in-situ lesions and in invasive carcinomas.  

Conclusions:

The results show that there is an inverse relationship between the extent of apoptosis and bcl-2 expression in breast lesions suggesting that its expression affects the regulation of apoptosis in them.