微波、噪声、振动条件对脑单胺类含量的影响

本文观察了急性或慢性微波、噪声、振动条件下小鼠、家兔脑单胺类递质含量的变化。结果表明,急性噪声条件下小鼠间脑和脑干5-HT含量明显升高,DA含量有升高趋势;急性振动时,家兔下丘脑NE含量明显升高。慢性微波照射条件下,小鼠全脑5-HT含量明显下降,慢性噪声作用下间脑和脑干DA、NE呈下降趋势,我们还初步分析了上述结果与临床微波作业人员的职业病症状间的关系。     

HPLC—ECD法分析中枢单胺类递质及其在免疫应答过程中的动态变化

建立快速、简便反相离子对高效液相色谱—电化学检测分析法,一次同时测定生物样品中单胺类递质(NE.E.DA、5—HT)及其前体氨基酸(Tyr、Trp)和主要代谢产物(HVA、5-HIAA共8种化合物。应用该法检测了SRBC免疫小鼠大脑皮质、间脑、脑干8种单胺类化合物的变化规律。免疫后第3天,皮质DA/HVA、间脑NE含量显著降低(P<0.05);第5天,间脑5-HT和脑干5-HIAA含量明显升高(P<0.05)。支持免疫应答可以影响中枢单胺类递质代谢的推论。     

全脑照射后对小鼠血脑屏障通透性和室管膜下区细胞增殖的影响

目的:探讨60Co-γ射线(Gy)全脑照射后对小鼠血脑屏障通透性和室管膜下区神经干细胞增殖的影响。方法:112只健康小鼠随机分成0 Gy组(对照组),5 Gy照射组(小剂量照射组),15 Gy照射组(中等剂量照射组)和30 Gy照射组(大剂量照射组),每组28只。各组随机取出7只分别于照射后1周和4周测定各组小鼠脑组织伊文思蓝的含量,7只观察室管膜下区的BrdU+细胞形态和数量。结果:(1)照射后1周,15 Gy组和30Gy组脑组织伊文思蓝含量明显升高(P<0.05),室管膜下区的BrdU+细胞数明显降低(P<0.05);(2)照射后4周,15 Gy剂量照射组脑组织伊文思蓝含量和室管膜下区的BrdU+细胞数均恢复到对照组水平(P>0.05),而30Gy剂量照射组脑组织伊文思蓝含量仍明显升高(P<0.05),BrdU+细胞数也未见恢复(P<0.05)。结论:全脑照射后血脑屏障破坏可能对室管膜下区的细胞增殖有进一步抑制作用。     

比较热休克蛋白A5和3-甲基腺嘌呤和介导小鼠脑缺血再灌注损伤的保护作用

目的:比较研究热休克蛋白A5(HSPA5)和3-甲基腺嘌呤(3-MA)介导小鼠脑缺血再灌注(I/R)损伤性自噬的保护作用。方法:成年雄性BALB/c小鼠30只,分为6组:sham组、缺血再灌注组(I/R组)、vehicle+I/R组、3-甲基腺嘌呤(3-MA)+I/R组、scramble siRNA+I/R组和HSPA5 siRNA+I/R组,每组5只。采用大脑中动脉阻塞(MCAO)60 min后再灌注24 h制作小鼠I/R模型。Vehicle+I/R组和3-MA+I/R将5μl 0.9%Na Cl或3-MA(30 mg/ml)在MCAO前30 min侧脑室注射;scramble siRNA+I/R group组和HSPA5 siRNA+I/R组将5μl scramble siRNA或HSPA5 siRNA(2μg/μl)在MCAO前24 h侧脑室注射。通过小鼠I/R后神经功能评分确定运动功能障碍程度。小鼠神经细胞内自噬体形成用透射电子显微镜测定,再灌注24 h缺血大脑皮层(LC3)-II/LC3-I表达用Western Blot方法检测。小鼠I/R后缺血大脑神经细胞损伤用Nissl染色进行观察。结果:HSPA5和3-MA可以影响I/R小鼠行为学改变并使脑缺血性损伤和神经症状加重(P0.05)。透射电子显微镜检测可见,sham组小鼠大脑皮层神经细胞形态正常,I/R组小鼠缺血大脑皮层神经元胞质中细胞器减少,形成自噬体。与sham组小鼠比较,I/R组小鼠缺血大脑皮层LC3-II蛋白表达水平显著增高(P0.05);3-MA+I/R或HSPA5 siRNA+I/R小鼠缺血大脑皮层LC3-II蛋白表达水平明显低于I/R小鼠(P0.05)。结论:HSPA5和3-MA在介导小鼠脑缺血再灌注损伤导致的自噬可发挥相同的保护作用,并且促进神经细胞凋亡。     

几种维生素对噪声暴露小鼠肝脏糖原含量和GPT活性变化的影响

小鼠噪声暴露(3.5KH_z,103dB)60min时,肝糖原含量.饱食鼠噪声组显著低于对照组(P<0.01),事先腹腔注入Vit B_(12)、C各组高于噪声组(P<0.05),接近对照组(P>0.05);饥饿鼠噪声组.Vit B_(12)和C组均高于对照组(P<0.05和P<0.01)。肝GPT活性:饱食鼠噪卢组显著高于对照组(P<0.01),而Vit B_(12) C和E各组与对照组无差异。饥饿鼠各组间比均无差导。说明Vit B_(12)C和E均有对抗噪声引起饱食鼠肝糖原含量下降和阻碍噪声所致饱食鼠肝GPT活性上升等作用。     

Neuroplastin65敲除通过调控中枢5-HT影响小鼠的焦虑样行为

目的:研究neuroplastin65(NP65)敲除对小鼠焦虑样行为的影响并初步探讨其机制。方法:成年NP65敲除(NP65~(-/-))小鼠和同基因型背景的野生型(wild type,WT)小鼠,通过Western Blot鉴定NP65~(-/-)小鼠;用旷场实验、探洞实验和埋珠实验检测小鼠的焦虑水平;ELISA检测小鼠海马、前额叶皮质和纹状体谷氨酸和5-羟色胺(5-hydroxytryptamine,5-HT)的含量;5-HT合成酶色氨酸羟化酶2(tryptophan hydroxylase 2,TPH2)免疫荧光染色观察小鼠脑干中缝核5-HT能神经元;Western Blot检测小鼠脑干TPH2的蛋白表达。结果:与WT小鼠相比,在旷场实验中,NP65~(-/-)小鼠进入中心区的次数显著减少(P0.001),中心区停留时间显著降低(P0.001),运动总距离和平均运动速度无显著变化。探洞实验中,NP65~(-/-)小鼠探洞次数显著增加(P0.001)、探洞时间增多(P0.05)、后肢竖立次数减少(P0.01)和排便次数增多(P0.001)。埋珠实验中,NP65~(-/-)小鼠掩埋潜伏期延长(P0.05),埋珠个数显著减少(P0.05)。神经递质检测结果显示:NP65~(-/-)小鼠海马、前额叶皮质和纹状体谷氨酸的含量不变,海马5-HT的含量显著减少(P0.05),但前额叶皮质和纹状体5-HT的含量没有显著性差异。免疫荧光染色观察到NP65~(-/-)小鼠脑干中缝核TPH2阳性细胞数目显著减少(P0.05)。Western Blot分析显示:NP65~(-/-)小鼠脑干TPH2的蛋白表达量显著降低(P0.05)。结论:NP65敲除导致小鼠表现出显著的焦虑样行为,可能与脑干中缝核TPH2表达减少而导致海马5-HT含量降低有关。     

血管内皮生长因子B在牦牛间脑和脑干相关组织的表达与定位

目的 了解牦牛间脑和脑干不同区域血管内皮生长因子B(VEGF-B)的表达及分布特征，探讨其与低氧适应性之间的关系。方法 选取5头健康牦牛，按照脑大体解剖结构进行分区并采集间脑和脑干相关组织，具体包括垂体、丘脑、下丘脑、延髓和脑桥5个部位。利用Real-time PCR、Western blotting及免疫组织化学技术对VEGF-B在牦牛间脑和脑干相关区域的表达与定位进行研究。结果 牦牛间脑和脑干相关组织中VEGF-B mRNA表达水平最高的部位为垂体，显著高于其他部位(P<0.05),其余表达水平依次为下丘脑、丘脑和延髓，脑桥表达量最低；牦牛间脑和脑干相关组织VEGF-B蛋白表达水平除丘脑高于下丘脑外，其余部位表达水平与mRNA表达趋势一致。免疫组织化学结果显示，与阴性对照组相比，VEGF-B蛋白阳性物质主要分布于上述各类细胞的胞质。其中，垂体中VEGF-B阳性着色主要表达于嗜酸性细胞中；丘脑和下丘脑中VEGF-B阳性着色主要集中于多形细胞；延髓和脑桥中VEGF-B阳性反应主要集中在神经元胞体中。结论 VEGF-B蛋白在牦牛间脑和脑干中均有表达，可能与其所担负的抗细胞凋亡、“生...     

手机电磁辐射对小鼠自由基影响的研究

目的 :研究经手机电磁辐射后的小鼠血清中SOD、MDA含量的变化 ,推断手机电磁辐射可能对人体造成的早期损害。方法 :应用分光光度比色法检测手机辐射后血清SOD、MDA含量 ,并与对照组进行统计学比较 ,同时电镜下观察大脑皮层、海马、小脑超微结构的变化。结果 :实验组小鼠血清SOD低于对照组 ,有显著性差异 (P <0 .0 5 ) ,而实验组MDA含量明显高于对照组 (P <0 .0 1) ;电镜下 ,实验组大脑皮层、海马、小脑超微结构与对照组相比较没有明显改变。结论 :手机辐射使小鼠体内自由基生成增多 ,对机体细胞造成早期损伤。     

OAZI-1 蛋白复合物在小鼠体内诱导特异性抗肿瘤效应的研究

目的:在实验动物的水平上分析来源于肿瘤细胞的鸟氨酸脱羧酶抗酶抑制因子-1(Ornithine decarboxylaseantizyme inhibitor-1,OAZI-1) 蛋白复合物能否在小鼠体内诱导特异性抗肿瘤效应。方法:用包被有OAZI-1 抗体的免疫磁珠从B16-F1 小鼠黑色素瘤细胞中分离OAZI-1 蛋白复合物,用此复合物免疫小鼠后, 再在小鼠皮下接种B16-F1 活细胞,然后观察接种瘤在小鼠体内的成瘤及生长状况。ELISA 法用于检测免疫小鼠血清中IFN-γ含量。乳酸脱氢酶释放实验(LDH)检测免疫小鼠脾脏淋巴细胞对B16-F1 细胞的杀伤效应。上述动物实验用原核表达纯化的OAZI-1 蛋白和PBS 免疫的小鼠作为对照。结果:与对照小鼠相比,接种OAZI-1 蛋白复合物的小鼠脾淋巴细胞(效应细胞)对B16-F1 黑素瘤细胞(靶细胞)具有更强的杀伤能力。在三种不同的效：靶比下(10 ：1、50 ：1、100 ：1),该组小鼠脾淋巴细胞对靶细胞的杀伤活性分别为46.2%、59.5% 和92.5%,显著性高于接种纯化OAZI-1 蛋白组(36.1%、26.8% 和45.9%)和接种PBS 组(24.6%、24.0% 和27.2%)小鼠脾淋巴细胞。此外,接种OAZI-1 蛋白复合物的小鼠血清中抗肿瘤细胞因子IFN-γ含量(538.3 pg/ ml)也显著性高于接种纯化OAZI-1 蛋白组(256.2 pg/ ml)和接种PBS 组(131.0 pg/ ml)小鼠。上述方法免疫的小鼠在皮下再接种B16-F1 活细胞后,免疫OAZI-1 蛋白复合物组小鼠成瘤率为40%,而PBS 组和纯化OAZI-1 蛋白组小鼠成瘤率为100%,且接种瘤在OAZI-1 蛋白复合物免疫小鼠体内生长更为缓慢。结论:从B16-F1 肿瘤细胞中分离的OAZI-1 蛋白复合物中可能含有肿瘤抗原,用此复合物接种小鼠能在实验动物体内诱导抗肿瘤免疫杀伤活性。     

大麻素受体激动剂花生四烯酰氯乙胺(ACEA)通过抑制炎症反应改善小鼠脓毒症相关性脑病

目的 探讨大麻素受体激动剂花生四烯酰氯乙胺(ACEA)对脓毒症相关性脑病(SAE)小鼠认知功能的影响。方法C57BL/6小鼠随机分成人工脑脊液(ACSF)组和脂多糖(LPS)组,通过采用LPS侧脑室注射构建SAE模型,小鼠脓毒症严重程度评分(MSS)和体质量变化判断小鼠的基本情况;行为学范式评估小鼠运动能力(旷场实验)、认知功能(情景前置性条件恐惧实验、 Y迷宫实验)。评估ACEA干预效果中,小鼠随机分为ACSF组、 ACEA干预的ACSF组、 LPS组和ACEA干预的LPS组,ACEA干预组给与1.5 mg/kg ACEA,实时定量PCR检测小鼠海马组织白细胞介素1β(IL-1β)、 IL-6、肿瘤坏死因子α(TNF-α)的mRNA表达水平;Western blot法检测小鼠海马组织IL-6、 TNF-α的蛋白表达水平;尼氏染色检测小鼠海马CA1区神经元损伤情况;行为学范式评估小鼠运动能力(旷场实验)、认知功能(情景前置性条件恐惧实验、 Y迷宫实验)。结果 侧脑室注射LPS 3 d后,小鼠存在显著认知功能障碍,证实SAE造模成功。与ACSF组相比,LPS组海马炎症因子IL-6、 TN...     

Autonomic regulation of insulin secretion is changed by pentobarbital in mice

It has been established that insulin secretion is regulated by autonomic nervous homeostasis. In the screen of plasma glucose level, anesthetized animals were widely used. However, effects of anesthetics on blood glucose remain unclear. In the present study, we compared the hypoglycemic action of ginseng that was induced by insulin secretion in mice between conscious and under anesthesia with pentobarbital. The hypoglycemic effect of ginseng was only produced in anesthetized BALB/c mice but not in the conscious mice. Similar results were also observed in C57BL/6 mice. However, the hypoglycemic action of ginseng failed to produce in anesthetized BALB/c mice received streptozotocin to induce type-1 like diabetes showing an insulin-dependent manner. The plasma insulin level in anesthetized BALB/c mice was markedly raised by ginseng but this effect was not observed in conscious mice. Blockade of muscarinic receptors by atropine inhibited ginseng-induced insulin secretion in anesthetized mice. Otherwise, the hypoglycemic action of ginseng was restored in conscious mice treated guanethidine at a sufficient dose to block sympathetic tone. In conclusion, the obtained results suggest that insulin secretion regulated by autonomic nervous homeostasis can be changed by pentobarbital through decrement in sympathetic tone to increase the insulin secretion induced by agent(s) via higher of parasympathetic tone. This finding is suitable to explain the critical hypoglycemia was not observed in subjects received ginseng.     

Post-exposure treatment with ginsenoside compound K ameliorates auditory functional injury associated with noise-induced hearing loss in mice

Noise-induced hearing loss (NIHL) is thought to primarily involve damage to the sensory hair cells of the cochlea via mechanical and metabolic mechanisms. Unfortunately, initial studies assessing the effectiveness of post-exposure treatment after hearing loss have yielded largely disappointing results. This study explored the effects of oral treatment with Korean red ginseng (RG) and with two bioavailable ginsenoside metabolites, ginsenoside Rh1 and ginsenoside compound K (GCK), in response to NIHL in a murine model. Pharmacological treatments began 24h after noise exposure and were continued once daily for 7 days. Central auditory function was evaluated using auditory middle latency responses, and cochlear function was determined based on transient evoked otoacoustic emissions. Additionally, cochlear hair cell morphology was investigated after noise exposure. Both Korean red ginseng and compound K reduced threshold shifts, central auditory function damage, and cochlear functional and morphological deficits. In contrast, treatment with ginsenoside Rh1 did not result in recovery of NIHL in mice. These results suggest that consumption of Korean red ginseng may facilitate recovery from noise-induced hearing loss. Furthermore, one of the active constituents in ginseng is likely ginsenoside compound K.     

Antidepressant-like effect of altered Korean red ginseng in mice

Altered Korean red ginseng has been used as a treatment for patients suffering from anxiety. We assessed whether red ginseng hydrolyzed by malted barley (HRG) and acetate-fermented red ginseng (ARG) would improve brain activity, by using forced swimming test (FST) in mice. The effect of the fluoxetine (a classical antidepressant), ginsenoside Rg3 (Rg3), red ginseng (RG), HRG, and the ARG groups for two weeks on the immobility time was significantly decreased in comparison with the control group (p<0.05). The immobility time of HRG and ARG in FST was lower than that of RG. The plasma level of glucose and total protein was significantly increased in the HRG and ARG group compared with the control group (p<0.05), whereas albumin, lactate dehydrogenase, creatine kinase, and blood urea nitrogen levels were not changed. In conclusion, altered Korean red ginsengs, HRG, and ARG therapy appeared to be effective in improving depression.     

大鼠下边缘皮质的皮质下端脑,间脑,脑干的传入投射

实验用ＨＲＰ导入大鼠下边缘皮质研究了皮质下端脑、间脑和脑干至下边缘皮质的传入投射。结果：皮质下端脑至下边缘皮质的传入主要来自同侧屏状核,背侧梨状内核,杏仁基底外侧核和杏仁基底内侧核,其次来自杏仁外侧核,杏仁皮质核等。     

Antidepressant-like Effect of Altered Korean Red Ginseng in Mice

Altered Korean red ginseng has been used as a treatment for patients suffering from anxiety. We assessed whether red ginseng hydrolyzed by malted barley (HRG) and acetate-fermented red ginseng (ARG) would improve brain activity, by using forced swimming test (FST) in mice. The effect of the fluoxetine (a classical antidepressant), ginsenoside Rg3 (Rg3), red ginseng (RG), HRG, and the ARG groups for two weeks on the immobility time was significantly decreased in comparison with the control group (p < 0.05). The immobility time of HRG and ARG in FST was lower than that of RG. The plasma level of glucose and total protein was significantly increased in the HRG and ARG group compared with the control group (p < 0.05), whereas albumin, lactate dehydrogenase, creatine kinase, and blood urea nitrogen levels were not changed. In conclusion, altered Korean red ginsengs, HRG, and ARG therapy appeared to be effective in improving depression.     

Increase of acetylcholine release by Panax ginseng root enhances insulin secretion in Wistar rats

The present study was designed to ascertain the effect of Panax ginseng root on plasma glucose and investigate the possible mechanisms for the effect. Ninety minutes after the oral administration of P. ginseng root to fasting Wistar rats, plasma glucose decreased in a dose-dependent manner. Simultaneous with the reduction in plasma glucose, an increase in the plasma level of insulin and C-peptide was also observed. Moreover, disruption of the available synaptic acetylcholine (ACh), using the inhibitor for choline uptake (hemicholinium-3), or the inhibitor for vesicular choline transport (vesamicol), abolished the metabolic actions of P. ginseng root. Conversely, physostigmine, at a concentration sufficient to inhibit acetylcholinesterase, enhanced the metabolic effect of P. ginseng root. It is possible that P. ginseng root mediates the release of ACh from nerve terminals to enhance insulin secretion. Blockade of the actions of P. ginseng root by 4-diphenylacetoxy-N-methylpiperdine methiodide (4-DAMP) suggested that the site of action is the muscarinic M(3) receptor. Taken together, the results suggest that P. ginseng root has the ability to increase the release of ACh from nerve terminals in rats so as to stimulate muscarinic M(3) receptors activity located in the pancreatic cells for the secretion of insulin, which in turn lower plasma glucose.     

The quaking mouse: regional variations in the content and protein composition of myelin isolated from the central nervous system

Myelin was isolated from six regions of the central nervous system of quaking and littermate control mice. The yield of myelin from the mutant animals ranged from 2% of the control level in the most rostral parts of the central nervous system to 16% in the spinal cord, with an intermediate value (10%) for the optic nerves; these results are in close agreement with the data of others obtained by quantitative electron microscopy. Electrophoretic analyses of myelin from the telencephalon, diencephalon, brain stem and spinal cord revealed a marked deficit of proteolipid and small basic proteins in all fractions from quaking mice; however, neither of these proteins exhibited clear rostro-caudal trends with respect to the ratio of protein contents between quaking and control mice.An hypothesis is proposed to account for the regional variations in the mutant's myelin deficit; it is suggested that the quaking mutation induces an arrest in myelinogenesis through a mechanism which is common to both oligodendroglial and Schwann cells.     

中药抗突变实验研究

本文从1984年起分别以C57纯种小鼠的骨髓细胞和人外周血淋巴细胞为实验材料，用公认的诱变剂环磷酰胺(CPP)诱发的姐妹染色单体互换(SCE)，微核(MN)升高作指标，对大量中草药进行实验，已筛选出部分补气药或相关物以及各种人参的有效成分或单体具有抗突变作用。