缺氧缺血性脑损伤新生大鼠脑干听觉诱发电位的动态变化

目的：探讨缺氧缺血性脑损伤新生大鼠脑干听觉诱发电位（BAEP）的动态变化。方法：30只出生13天的新生SD大鼠随机分为对照组和缺氧缺血组,每组15只。将出生后第13天新生大鼠结扎左侧颈总动脉后,置于含氧8％的低氧环境中3h建立缺氧缺血性脑损伤动物模型。出生后第14、20、27、37、47、57、67和77天检测大鼠的BAEP。结果：缺氧缺血组新生大鼠3h缺氧复氧1h后,表现为向左侧旋转,爬行时右侧后肢呈现拖步。出生后第14～47天缺氧缺血组大鼠BAEP波峰潜伏期（PL）和波峰间潜伏期（IPL）较对照组显著延长（P〈0．05或P〈0．01）,这些鼠的BAEP的PL及IPL均随年龄的增加而缩短,并且鼠可发育成熟。结论：缺氧缺血性脑损伤新生大鼠BAEP有显著的可逆性异常变化,其PL和IPL是动态监测与客观评价新生期缺氧缺血性脑损伤和听觉障碍的灵敏指标。     

黄芪对缺氧缺血性脑损伤新生鼠脑组织Nogo-A蛋白表达的作用及意义

目的探讨Nogo-A蛋白在缺氧缺血性脑损伤新生鼠脑组织中的表达特点及黄芪对其表达的影响方法 7日龄Wistar新生大鼠72只,清洁级,雌雄不限,平均体重10-16g,随机分为假手术组,HIBD组,黄芪治疗组,每组24只。分离左侧颈总动脉并结扎,吸入92%N2+8%O2氮氧混合气体行缺氧处理2h,制成HIBD模型。假手术组只作分离左侧颈总动脉而不结扎,不作缺氧处理,不给与药物处理。黄芪治疗组于建模成功后立即及3h腹腔注射0.5ml/只,假手术组及HIBD组大鼠腹腔注射等量生理盐水,上述各组均于术后6h、12h、24h、72h随机抽取6只,于麻醉下处死取脑组织,HE染色观察脑组织病例变化,免疫组化法检测Nogo-A蛋白表达情况。结果与假手术组相比,HIBD组术后脑组织Nogo-A蛋白表达增高,但Nogo-A蛋白随各时间点有先上升后下降的趋势,差异有统计学意义(P0.05),与HIBD组相比,黄芪治疗组各时间点Nogo-A蛋白,差异有统计学意义(P0.05)。结论提示黄芪可下调缺氧缺血性损伤大鼠脑组织Nogo-A蛋白的表达,对神经元和神经纤维具有保护作用。     

高压氧对缺氧缺血性脑损伤新生大鼠TLR-4信号通路的影响

目的探讨高压氧对缺氧缺血性脑损伤新生大鼠TLR-4信号通路的影响。方法选取36只7日龄SD新生大鼠,随机分为3组:假手术组(n=12),缺氧缺血性脑损伤组(n=12),高压氧治疗组(n=12)。采用Rice法制作新生大鼠缺氧缺血性脑损伤模型,假手术组大鼠分离左颈总动脉后不结扎直接缝合皮肤,高压氧治疗组大鼠在模型制作成功后,给予高压氧治疗,1 h/d,连续7 d。测定各组大鼠体质量增长率与左/右脑重量比值;应用TUNEL法检测脑组织神经细胞凋亡情况,Western Blot方法与real time PCR方法分别检测各组新生大鼠脑组织TLR-4、MyD88与NF-κB蛋白与mRNA的表达。结果高压氧治疗可使缺氧缺血性脑损伤新生大鼠的体质量增长率和左/右脑重量比值增加(P0.05)。与假手术组相比,缺氧缺血性脑损伤组新生大鼠脑组织的神经细胞凋亡数目、Toll样受体-4(TLR-4)、髓样分化因子(MyD88)与核转录因子-κB (NF-κB)蛋白与mRNA的表达均显著升高(P0.01);与缺氧缺血性脑损伤组相比,高压氧治疗组新生大鼠脑组织的神经细胞凋亡数目、TLR-4、MyD88与NF-κB蛋白与mRNA的表达均显著降低(P0.01)。结论高压氧减轻缺氧缺血性脑损伤新生大鼠的脑组织损伤可能与抑制TLR-4信号通路相关。     

缺氧缺血性脑损伤对新生大鼠发育期间额叶皮质突触质量的影响

用 7日龄 Wistar大鼠制成缺氧缺血脑损伤模型 ,从生后 7d到成年分 8个时间段对突触的质量进行了长期跟踪研究。结果显示 :脑损伤后鼠的额叶皮质神经元及神经纤维的密度明显降低 ( P<0 .0 5 ,P<0 .0 1) ;突触的体视学参数表面积密度和面数密度降低 ;突触后膜致密层变薄 ( P<0 .0 5 ,P<0 .0 1)。实验结果提示 :新生鼠脑缺氧缺血损伤后额叶皮质内突触质和量的改变是影响智力发育的重要因素之一 ;同时还表明神经纤维及突触有代偿性。这些是实行早期干预改善脑功能的形态学基础     

大鼠缺氧缺血性脑损伤脑组织中脑红蛋白基因表达的变化

20 0 0年 ,德国Ｂｕｒｍｅｓｔｅｒ等[1] 首次报道了人和小鼠脑内存在特异的功能上类似于肌红蛋白的携氧球蛋白———脑红蛋白 (ｎｅｕｒｏｇｌｏｂｉｎ) ,可特异性的为脑供氧 ,从而对脑缺氧的研究提供了全新思路[2 ] ,并已在国际范围内掀起重新认识和研究脑缺氧的热潮。我们实验室一直在关注脑缺氧损伤的分子机制研究 ,并获得了人脑红蛋白全长ｃＤＮＡ序列和大鼠脑红蛋白基因编码区的ｃＤＮＡ序列[3 ,4] ,随后根据此序列设计引物 ,采用逆转录 聚合酶链反应 (ＲＴ ＰＣＲ)技术对大鼠缺氧缺血性脑损伤时脑组织中脑红蛋白ｍＲＮＡ水平的变化进…     

红景天苷对缺氧缺血性脑损伤大鼠海马神经元自噬的影响

目的:探讨红景天苷(Sal)对缺氧-缺血(H/I)大鼠海马神经元自噬的影响。方法:36只成年雄性SD大鼠分为假手术组(Sham)、H/I组(H/I)、H/I联合Sal治疗组(H/I+Sal),利用双侧颈总动脉结扎及低氧处理制备H/I模型,通过腹腔注射给予大鼠Sal治疗3 d,Morris水迷宫实验检测大鼠学习记忆能力,TUNEL染色检测海马神经元凋亡,Western Blot检测微管相关蛋白1轻链3(LC3)的表达。结果:与sham组相比,H/I大鼠学习记忆能力受损明显(P 0. 05),海马细胞凋亡增加明显(P 0. 05),LC3-II/LC3-I比值增加(P 0. 05);经过Sal治疗后,与H/I大鼠相比,Sal治疗组大鼠学习记忆功能得到明显改善(P 0. 05),凋亡细胞减少(P 0. 05),LC3-II/LC3-I比值降低(P 0. 05)。结论:Sal能够通过抑制H/I模型大鼠海马神经元自噬减轻凋亡,发挥神经保护作用。     

围产期缺氧缺血性脑损伤大鼠神经细胞凋亡及巴曲酶神经保护作用的研究

围产期窒息所致缺氧缺血性脑病(HIE)是新生儿期危害最大的常见病,常导致新生儿死亡,存活者可遗留脑性瘫痪、癫痫、智力低下等严重的神经系统后遗症.本文通过夹闭孕足月大鼠双侧子宫动脉复制宫内缺氧缺血性脑损伤(HIBD)动物模型,检测不同缺氧缺血再灌注时间点宫内HIBD仓鼠神经细胞凋亡数量,应用工酶治疗HIBD仔鼠,研究其神经保护作用.     

缺氧缺血性脑损伤新生大鼠海马突触体素的表达及当归调控作用

目的探讨脑缺氧缺血对新生大鼠海马齿状回突触体素表达的影响及当归注射对其表达的调控作用。方法取7日龄健康SD新生大鼠33只,随机分为对照组、缺氧组和当归组各11只。缺氧组和当归组新生大鼠在无菌环境下结扎左侧颈总动脉,术后护理2 h后置于三气培养箱持续缺氧2 h,制作新生鼠缺氧缺血性脑损伤（HIBD）模型,对照组仅行假手术,不结扎左侧颈总动脉、不缺氧。术后第8 d开始,缺氧组和对照组大鼠经腹腔注射生理盐水（8ml/kg）,连续7 d;当归组用等量当归注射液（250 g/L）代替生理盐水。于生后第22 d取大鼠脑组织,常规石蜡包埋、经海马切片,行突触体素（SY）免疫组化染色,图像分析海马齿状回SY阳性细胞的积分光密度（IOD）值。结果缺氧组大鼠海马齿状回SY阳性细胞的IOD值较对照组降低,而当归组SY阳性细胞的IOD值较缺氧组增高。结论脑缺氧缺血可降低新生大鼠海马齿状回SY的表达,而当归注射液对缺氧缺血性脑损伤新生大鼠可能具有保护作用。     

高压氧降低缺氧缺血性脑损伤大鼠脑组织内caspase-3表达

目的 探讨高压氧(HB0)对缺氧缺血新生大鼠神经细胞凋亡及easpase-3表达的影响.方法 建立缺氧缺血性脑损伤(HIBD)大鼠模型,将其分为HIBD组及HBO处理组,并设立假手术组(n=24).观察3组大鼠不同时间脑组织病理学改变,同时用免疫组化的方法检测3组大鼠脑组织内ca8pase-3蛋白表达的动态变化.结果 HIBD组18、24、48和96 h时间点海马及皮层区ca8pase-3蛋白的表达均明显高于假手术组(P相似文献   

新生大鼠缺氧缺血性脑损伤对脾bcl-2/bax/fas/fasL mRNA的影响

目的 通过检测脾淋巴细胞凋亡基因bcl-2/bax/fas/fasL mRNA表达水平,探讨缺氧缺血性肭损伤(hypoxicischemic brain damage,HIBD)脾细胞凋亡基因的变化与缺氧时间的关系,以此提供HIBD对于免疫功能影响的理论基础.方法 建立新生大鼠HIBD模型测定脾bcl-2/bax/fas/fasL mRNA 的相对表达量.结果 bcl-2处理组相对表达量低于对照组(P＜0.01).bax处理组相对表达量高于对照组(P＜0.01).bcl-2/bax处理组比值低于相应对照组(P＜0.05).fas处理纽相对表达量高于对照组(P＜0.01).fasL 1 h和6 h处理组相对表达量高于对照组,12 h之后则对照组远高于处理组.fas/fasL对照组较分散,除6 h组外其余组均小于1,处理组较集中,除24 h外均大于1.结论 HIBD可促进脾细胞促凋亡基因的表达,减少抑凋亡基因表达;HIBD发生6～12 h是促凋亡基因和抑凋亡基因高峰表达期;HIBD加强了fas/fasL的作用.     

Effects of ultra-marathon on circulating DNA and mRNA expression of pro- and anti-apoptotic genes in mononuclear cells

We investigated the effects of an ultra-marathon on cell-free plasma DNA as well as on mRNA expression of pro-apoptotic (Bax, Bad), anti-apoptotic (Bcl-2) and cell-protective (Hsp70, Hsp27 and Hsp32) genes in mononuclear blood cells (MNCs). Blood samples were drawn from 14 athletes before and immediately after 6-h run. In addition, blood samples were also collected and analyzed 2 and 24 h after the end of the run. Levels of plasma DNA were significantly increased immediately after the marathon (P < 0.001) and were still higher 2 h later (P < 0.005), but significantly lower than those immediately after the race (P < 0.05). Cell-free plasma DNA returned to pre-race levels 24 h after the run. mRNA expressions of Hsp70, Hsp32 and Bax significantly increased in MNCs after the race, whereas Hsp27 and Bad mRNA expression levels showed no significant changes. Bcl-2 expressions decreased immediately after the race (P < 0.001), but increased in the 24 h later (P < 0.05). We conclude that apoptotic ladders of cell-free DNA following exhaustive exercise originate from apoptotic cells and that not only skeletal muscle cells but also leukocytes contribute to this phenomenon. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

缺氧缺血性脑损伤与凋亡的进展

Apoptosis is one of the most important causes, which results in the central neuronal system complication in hypoxic- ischemic brain damage (HIBD). Apoptosis occurs in the developing brain more than in the developed brain. Apoptosis can last several weeks and may be inverted its pathology by appropriate therapy. Caspase inhibitor, neurotrophic factors, anti-apoptosis gene Bcl-2, mild hypothermia, and early intervention play important roles in promoting neuronal cell survival and preventing from apoptosis through different mechanisms. It may be a new way for rehabilitation of HIBD.     

瑞舒伐他汀预处理对局灶性脑缺血/再灌注损伤大鼠Bcl-2和Bax表达的影响

目的:探讨瑞舒伐他汀预处理对大鼠脑缺血/再灌注损伤脑组织Bcl-2和Bax表达的影响。方法:48只SD大鼠随机分为假手术组(Sham组)、脑缺血/再灌注组(I/R组)、溶剂对照组(Vehicle组)和瑞舒伐他汀预处理组(Ros组)。Ros组在模型制备前用瑞舒伐他汀5 mg/kg/d连续灌胃10 d,1次/d;Vehicle组用相同体积的生理盐水连续灌胃10 d。线栓法制作大鼠大脑中动脉脑缺血/再灌注模型,于缺血2 h后再灌注24 h后行神经功能评分,断头取脑,免疫组织化学法和RT-PCR法观察脑组织Bcl-2和Bax表达水平的变化。结果:大鼠脑缺血/再灌注损伤后,Bcl-2和Bax显著增多;与I/R组和Vehicle组相比,Ros组神经功能改善,Bcl-2的表达显著上调,Bax的表达显著下调,Bcl-2/Bax比值显著升高,差异均有统计学意义(P<0.05)。结论:瑞舒伐他汀预处理对大鼠脑缺血/再灌注损伤有神经保护作用,其机制可能与上调脑组织中Bcl-2的表达,下调Bax的表达,Bcl-2/Bax比值增加,从而抑制胞凋亡有关。     

Changes in c-fos mRNA in the neonatal rat brain following hypoxic ischemia

We used quantitative in situ hybridization to study changes in the expression of c-fos following hypoxic-ischemia (H-I) in the neonatal rat brain. 7-day-old rat pups were subjected to a unilateral ligation of the common carotid artery followed by a 2 h 15 min hypoxic period (7.7% O₂ in N₂). This resulted in the expected ipsilateral infarction of cortex, lateral hippocampus, lateral-superior aspects of the striatum and the white matter of the corpus callosum. Brain damage was not seen in the contralateral hemisphere subjected only to hypoxia. c-fos mRNA levels increased in the contralateral hemisphere immediately after the hypoxia and had returned towards normal levels 2 h thereafter. In the ipsilateral hemisphere, the expression of c-fos was delayed but very marked at 2 h. Animals subjected only to hypoxia showed little or no increase in c-fos mRNA. Thus the earliest recorded increase in c-fos after hypoxic ischemia, which occurred on the non-ischemic, contralateral side, may represent a generalized response to a more localized insult.     

Effect of curcumin on Bcl-2 and Bax expression in nude mice prostate cancer

Prostate cancer is a common malignant tumor in urinary system. Curcumin has curative effect on many kinds of cancers and can inhibit prostate cancer (PC)-3 cells proliferation. This study aimed to explore the curcumin induced prostate cancer cell apoptosis and apoptosis related proteins Bcl-2 and Bax expression. PC-3 cells were injected subcutaneously to the nude mice to establish the tumor model. The nude mice were randomly divided into group C (normal saline), group B (6% polyethylene glycol and 6% anhydrous ethanol), group H, M, L (100 mg/kg, 50 mg/kg, and 25 mg/kg curcumin). The tumor volume was measured every 6 days to draw the tumor growth curve. The mice were killed at the 30^th day after injection to weight the tumor. TUNEL assay was applied to determine cell apoptosis. Immunohistochemistry was used to detect Bcl-2 and Bax expression. The tumor volume and weight in group H, M, L were significantly lower than the control group (C, B) (P<0.05), and the inhibitory rate increased following the curcumin dose increase. Compared with the control group, Bcl-2 expression in group H, M, L gradually decreased, while Bax protein expression increased (P<0.05). The cell apoptosis rate showed no statistical difference between group B and C, while it increased in curcumin group H, M, and L (P<0.05). Curcumin could inhibit PC-3 growth, decrease tumor volume, reduce tumor weight, and induce cell apoptosis under the skin of nude mice by up-regulating Bax and down-regulating Bcl-2.     

Bcl-2、Bax在功能失调性子宫出血子宫内膜中的表达

目的 研究凋亡调节蛋白Bcl-2、Bax在功能失调性子宫出血(功血)的子宫内膜中的表达.方法 采用免疫组织化学(免疫组化)方法,利用链霉素抗生物素蛋白--过氧化物酶法(SP)检测40例功血患者子宫内膜及40例子宫肌瘤标本的子宫内膜组织中Bcl-2、Bax基因的表达情况.结果 (1)Bel-2基因在正常月经周期子宫内膜组织的表达呈明显周期性变化,差异具有统计学意义(P<0.05).(2)Bcl-2基因随子宫内膜增生程度增加而表达逐渐增强,差异具有统计学意义(P<0.05).(3)Bax在正常月经周期子宫内膜组织的表达呈阳性.(4)Box随子宫内膜增生程度增加而表达逐渐减弱,差异具有统计学意义(P<0.05).结论 Bcl-2的过度表达和Bax的低表达可抑制子宫内膜细胞凋亡,使子宫内膜增生甚或不典型增生.与功能失调性子宫出血有密切关系.     

罗布麻颗粒对化疗型卵巢早衰大鼠Bax和Bcl-2蛋白表达的影响

目的： 探讨罗布麻颗粒对化疗型卵巢早衰大鼠Bax 和Bcl-2 蛋白表达的影响。方法：性成熟SD雌鼠随机分 为对照组、造模组,腹腔注射（ 顺铂1.5 mg/kg）制备卵巢早衰模型,随机分为卵巢早衰模型组和罗布麻颗粒高、 中、低剂量治疗组。H-E 染色观察卵巢形态;ELISA 检测各组大鼠血清Bax、Bcl-2 和雌二醇、促黄体生成素（LH）、 促卵泡激素（FSH））的含量,免疫组织化学法检测卵巢Bax 与Bcl-2 蛋白表达。结果：卵巢早衰模型组大鼠中初 级卵泡减少,闭锁卵泡增多,罗布麻颗粒高剂量治疗组大鼠可见初级卵泡,闭锁卵泡较卵巢早衰模型组减少;卵 巢早衰模型组大鼠血清中雌二醇及Bcl-2 含量降低,FSH、LH、Bax 的含量升高; 各药物治疗组雌二醇、Bcl-2 含 量升高,FSH、LH、Bax 的含量降低;Bax 蛋白主要表达在颗粒层细胞及卵泡液中,卵巢早衰模型组大鼠较对照 组表达显著升高, 各药物治疗组在不同程度上能够降低其表达;Bcl-2 蛋白在各组主要表达在颗粒层细胞中,卵 巢早衰模型组大鼠较对照组表达显著下降, 各药物治疗组在不同程度上能够升高其表达。结论：顺铂能够导致雌 性大鼠血清中性激素水平的紊乱及卵巢组织形态的改变,可能与细胞凋亡因子Bax 与Bcl-2 在血清及卵巢组织中 表达水平的高低有关;罗布麻颗粒可能通过下调Bax 蛋白、上调Bcl-2 蛋白从而改善顺铂致卵巢早衰大鼠的卵巢 功能。     

硼替佐米对神经母细胞瘤细胞Bax/Bcl-2蛋白表达的影响

目的:了解应用硼替佐米对人神经母细胞瘤的治疗效果及其可能机制。方法:取对数生长期人神经母细胞瘤SHSY5Y细胞株,采用四甲基偶氮唑盐比色法测定其药物对细胞增殖的影响并计算对细胞作用的IC_(50)值,Hoechst-33258染色观察细胞凋亡形态,免疫印迹检测Bax、Bcl-2表达。结果:20nmol/L硼替佐米可引起细胞发生肿胀、变圆等细胞凋亡等形态学变化;作用24 h后引起SH-SY5Y细胞中Bax蛋白表达水平增高,Bcl-2蛋白表达水平下降,Bax/Bcl-2比率较空白组增高;硼替佐米抑制神经母细胞瘤的增殖作用随着其浓度的升高而更加明显。结论:硼替佐米通过线粒体途径诱导细胞凋亡从而发挥抑制人神经母细胞瘤增殖的作用。     

高压氧对新生大鼠缺氧缺血性脑损伤后在体神经干细胞的影响

目的： 新生鼠的神经生发区-室管膜下区（SVZ） 和海马齿状回（DG）聚集了多种不同发育阶段神经干细胞（NSCs），它们可分化产生新的神经元和胶质细胞。本研究探讨缺氧缺血性脑损伤（HIBD）对脑生发区NSCs的损伤及高压氧（HBO）对此损伤的影响。从在体NSCs 探讨HBO对新生大鼠HIBD的保护作用机制。 方法： 新生7 d龄SD大鼠随机分为4组：① 正常对照组（CON，n=16）；② HIBD 模型组（HIBD，n=16）；③ 高压空气组（HBA，n=16）；④HBO治疗组（n=16）。Rice法复制HIBD模型，并予HBA或HBO治疗，每天1次共7 d。BrdU免疫组化显示在体NSCs。并取损伤侧脑SVZ区组织，体外NSCs培养并进行神经干细胞球计数。 结果： HIBD组生发区在体BrdU 阳性细胞和体外培养的神经干细胞球数目明显少于对照组。HBO组SVZ区的BrdU阳性细胞增多；体外培养的神经干细胞球增多。HBA组增加不明显。 结论： 新生大鼠HIBD后生发区NSCs减少，HBO治疗可以减轻HIBD后NSCs的死亡。HBA治疗无明显作用。     

Immunohistochemical expression of Bax and Bcl-2 in penile carcinoma

There is a complex interplay between the pro-apoptotic Bax and anti-apoptotic Bcl-2 family of proteins and the tumor suppressor gene p53. The pathogenic role of Bax and Bcl-2 protein expression in penile carcinomas has not previously been investigated. We examined Bax and Bcl-2 expression in verrucous (VC) and squamous cell carcinoma (SCC) of the penis. Herein we also present a concise review of p53, Bcl-2/Bax ratios, and their relationship to apoptosis. Fourteen cases of penile carcinoma, including 7 VC and 7 well-differentiated SCC, were analyzed for Bax and Bcl-2 expression by immunohistochemical analysis of paraffin embedded archived tissues. The number of positively staining tumor cells was enumerated per 100 tumor cells within non-overlapping high power fields. The Bax immunoreactivity was similar in VC (19+/-3%) and well-differentiated SCC (15+/-4%) (p = 0.69). The expression of Bcl-2 protein was significantly higher in well-differentiated SCC (69+/-12%) compared to VC (36+/-14%) (p = 0.04). The mean Bcl-2/Bax ratio was significantly lower in VC (1.89) compared to well-differentiated SCC (4.6) (p = 0.05). These findings indicate that penile VC and SCC are immunophenotypically distinct. Bax expression is comparable in verrucous and low-grade squamous cell carcinomas, but Bcl-2 expression of Bcl-2 is significantly higher in the squamous cell carcinomas.