微波辐射对小鼠脑内乙酰胆碱含量及胆碱乙酸转移酶活性的影响

用频率为2450MHz功率密度为10mW/Cm~2(WBASAR约11.4W/kg)的微波(连续波)对置于微波暗室内的昆明种雄性小鼠急性全身照射1小时后,立即按常规方法断头,取脑,制成样品,然后用放射免疫测定法测量小鼠脑内乙酰胆碱(ACh)含量及胆碱乙酰转移酶(ChAT)活性。结果表明:照射组的ACh含量为11.6±1.4pmol/mg(脑鲜重),ChAT活性为45.4±8.7pmolACh/min.mg(脑鲜重);而对照组的分别为16.0±2.1pmol/mg和61.0±13.8pmolACh/min.mg。证明微波照射后可引起动物脑内ACh水平和ChAT活性下降,提示微波辐射对中枢胆碱能系统确有不利影响。     

针刺对去卵巢大鼠脑内胆碱乙酰转移酶基因表达的影响

本工作旨在探讨雌激素对脑内乙酰胆碱生成的影响和电针刺激“足三里”穴对去卵巢大鼠脑内乙酰胆碱生成的调整作用。实验选用成年Wistar雌性大鼠,将动物分为正常对照组(INT)、去卵巢组(OVX)和去卵巢针刺组(OVX AC)。用放射免疫分析方法测定血中雌二醇含量,采用RT-PCR方法获得大鼠脑内胆碱乙酰转移酶(ChAT)mRNA的逆转录表达产物——cDNA,用琼脂糖凝胶电泳方法检测,并通过原位杂交方法观察海马ChAT mRNA阳性神经元的表达,然后用计算机图像分析系统进行统计分析。实验结果显示：去卵巢组大鼠体内雌激素水平明显降低,脑内ChAT mRNA的RT-PCR产物和海马ChAT mRNA阳性表达产物的平均面积、平均积分光度值均明显减少,与对照组和针刺组比较有显著性差异;去卵巢针刺“足三里”穴组与去卵巢组相比,大鼠血中雌激素水平明显升高,脑内ChAT mRNA RT-PCR产物明显增多,海马的ChAT mRNA表达阳性神经元增多。以上结果提示：脑内ChAT基因表达与体内雌激素水平有密切关系,去卵巢后针刺“足三里”穴对ChAT的调节作用可能是针刺增强脑内乙酰胆碱含量的机制之一。     

花生油对2型糖尿病大鼠海马CA1区神经生长因子及胆碱乙酰转移酶表达的影响

目的通过观察2型糖尿病大鼠海马CA1区神经生长因子（NGF）和胆碱乙酰转移酶（ChAT）表达的改变,研究花生油对2型糖尿病大鼠海马神经元NGF及ChAT表达的影响,探讨花生油在防治糖尿病脑病中的作用。方法 60只健康雄性SD大鼠随机分为4组：正常对照组（C组）、2型糖尿病组（T2DM组）、2型糖尿病给予2 mL花生油组（T2DM＋2 mL组）及2型糖尿病给予5 mL花生油组（T2DM＋5 mL组）。其中C组给予正常饮食,糖尿病组大鼠给予高脂饮食喂养,2个月后,按25 mg/kg体质量腹腔注射链脲佐菌素（STZ）制成2型糖尿病模型,T2DM组、T2DM＋2 mL组及T2DM＋5 mL组大鼠继续给予高脂饮食。糖尿病造模1个月后处死全部大鼠,行脑冰冻切片,用免疫组织化学方法检测各组大鼠海马CA1区NGF和ChAT的表达。结果 （1）T2DM组大鼠海马CA1区NGF表达比C组明显降低（P〈0.05）,T2DM＋2 mL组及T2DM＋5 mL组大鼠海马CA1区NGF表达均明显高于未给予花生油的T2DM组（P〈0.05）。（2）T2DM组大鼠海马CA1区ChAT表达显著低于C组（P〈0.05）,T2DM＋2 mL组和T2DM＋5 mL组大鼠海马CA1区ChAT表达均明显高于未给予花生油的T2DM组（P〈0.05）。结论 2型糖尿病大鼠海马CA1区神经生长因子表达降低,胆碱能神经元数量减少,这可能是2型糖尿病脑病发生的原因之一。花生油能增加2型糖尿病大鼠海马区内神经生长因子表达,促进胆碱能神经元存活,表明花生油具有一定的保护大鼠糖尿病脑病的作用。     

人红细胞内乙酰胆碱/胆碱活度的微电极测量及临床意义

本文采用新发展的乙酰胆碱/胆碱(ACh/Ch)离子选择微电极对人体单个红细胞内的ACh/Ch活度进行了测量。实验对象是25位正常人及8位躁狂症病人的红细胞。正常人红细胞内ACh/Ch的活度值为24.O±8.3×10^-6mol/L,而躁狂症病人的红细胞内ACh/Ch的活度值为98.3±41.4×10^-6mol/L,是正常人的4倍。本文对ACh/Ch离子选择微电极的结构和特性参数亦作了描述。     

微波辐射对药用植物生长发育的影响

微波辐射育种取得初步成果：适量微波辐射能提高药用植物种子的发芽率,缩短发芽历期,提高芽的的生长速度。     

微波辐射对家兔血清蛋白的影响

本文报道微波辐射对家兔血清蛋白及血沉的影响。使用微波理疗机对家兔睾丸局部照射１５分钟，温度４２℃。在照射前４天和照射后７天、１４天、５７天从心脏取血，进行血沉、总蛋白、白蛋白、球蛋白的测定。结果表明，照后７天，总蛋白、白蛋白、球蛋白均有显著下降，１４天后有所回升，５７天后基本恢复照前水平。照后７天的血沉只有少许改变，可见微波辐射的组织和器官扔直接作用外，还对机体血浆蛋白质代谢产生一定影响。     

微波辐射对药用植物生长发育的影响

微波辐射育种取得初步成果:适量微波辐射能提高药用植物种子的发芽率,缩短发芽历期,提高芽的的生长速度。     

脑源性神经生长因子对老龄鼠智能影响的探讨

目的研究脑源性神经营养因子（bDNF）改善痴呆老龄鼠学习记忆的作用机制;胆碱乙酰转移酶活性（ChAT-IR）、胆碱酯酶（AchE）活性的影响。方法利用海马内注射bDNF及免疫组织化学技术,观察给药前后各组迷宫试验,胆碱乙酰转移酶（ChAT）、胆碱酯酶（AchE）活性的变化。结果治疗组学习记忆能力有明显改变（P〈0.05）。与对照组比较ChAT-IR表达明显增加（P〈0.01）,AchE活性明显增强（P〈0.01）。结论bDNF使胆碱乙酰转移酶活性（ChAT）、胆碱酯酶（AchE）活性增强;对老龄鼠的智能有改善作用。     

17β-雌二醇对肺组织磷酸胆碱胞苷酰转移酶的影响

目的 :在离体肺组织培养模型上观察 1 7β 雌二醇 (1 7β estradiol,E2 )对肺表面活性物质主要成分磷脂酰胆碱合成的限速酶—CTP :磷酸胆碱胞苷酰转移酶 (CCT)活性的影响并探讨其作用机制。方法 :采用无血清成年大鼠肺组织培养 ,提取标记有1 4C的反应产物CDP 胆碱 ,液闪测定1 4C放射性强度 ,反映CCT活性。结果 :① 1× 1 0 - 7～ 3×1 0 - 6 mol/L的E2 以剂量依赖方式提高CCT活性 ;②用 3× 1 0 - 6 mol/L的E2 处理肺组织 2h、4h、8h和 1 6h ,在 8h时微粒体CCT活性显著高于对照组 (P <0 .0 1 ) ,1 6h酶活性进一步升高 ;③H 7和W 7可分别抑制 3× 1 0 - 6 mol/L的E2 促CCT活性的效应。结论 :E2 可促进成年大鼠CCT活性 ,其细胞内信号转导途径与蛋白激酶C和钙调蛋白有关。     

乙酰胆碱对自然杀伤细胞活性的影响

目的:观察乙酰胆碱(ACh)对自然杀伤(NK)细胞活性的影响,并初步探讨其作用的受体机制.方法:根据不同的实验目的,选择ACh、胆碱能受体激动剂和拮抗剂分别作用于NK细胞,以乳酸脱氢酶(lactate dehydrogenase,LDH)自然释放法检测不同实验条件下NK细胞杀伤肿瘤靶细胞(Yac)的活性.结果:ACh、M受体激动剂毛果芸香碱和N受体激动剂烟碱在10-10～10-6mol/L浓度范围内都能显著抑制NK细胞杀伤肿瘤细胞的活性.M受体拮抗剂阿托品(10-8和10-7mol/L)能完全阻断同浓度ACh抑制NK细胞活性的作用;但N受体拮抗剂筒箭毒碱(10-8和10-7mol/L)不能阻断同浓度ACh抑制NK细胞活性的作用.结论:ACh可抑制NK细胞对肿瘤细胞的杀伤作用,此作用主要由淋巴细胞上的M受体和N1受体介导.     

ACETYLCHOLINE,CHOLINE AND CHOLINE ACETYLTRANSFERASE ACTIVITY IN THE DEVELOPING BRAIN OF NORMAL AND HYPOTHYROID RATS1

Abstract— Acetylcholine, choline and choline acetyltransferase activity were measured in the whole brains of normal and hypothyroid rats during development. At 1 day postpartum, brain acetylcholine was 73 per cent of adult levels. Propylthiouracil-induced hypothyroidism up to age 20 days did not alter brain acetylcholine concentrations, but at 30 days resulted in significantly decreased levels. At day 1, brain choline was 20 per cent higher than adult levels and decreased between days 8 and 10. In hypothyroid rats this phenomenon did not occur until days 15–20. At day 1 postnatally, choline acetyltransferase activity was only 7 per cent of adult levels, then between days 5 and 20 rose to 77 per cent of adult levels. Beginning at day 8, hypothyroidism resulted in significantly decreased enzyme levels. This effect could be reversed at day 17 by concurrent tri-iodothyronine substitution therapy. In hypothyroid rats, maximum brain choline acetyltransferase activity was 30 per cent less than normal adult levels.     

EFFECTS OF BOTULINUM AND TETANUS TOXINS ON CHOLINE ACETYLTRANSFERASE ACTIVITY IN SKELETAL MUSCLE IN THE MOUSE

Abstract— The effects of botulinum and tetanus toxins on the activity of choline acetyltransferase present in the motor nerve terminals of fast and slow skeletal muscle in the mouse were investigated. There was no change in the activities of choline acetyltransferase in either muscle after the injection of botulinum toxin but tetanus toxin caused a rise in the activity of the enzyme in fast muscle. Botulinum toxin is known to inhibit the release of acetylcholine and whilst neuromuscular transmission is blocked the motor nerves sprout and form new end-plates. Tetanus toxin has been shown to cause hyperactivity of motor neurons. The nerve growth caused by the botulinum toxin did not result in increased choline acetyltransferase levels in the muscles, whereas the synaptic hyperactivity caused by tetanus was associated with increased enzyme levels.     

CHOLINE ACETYLTRANSFERASE CONTENT IN DISCRETE REGIONS OF THE RAT BRAIN STEM

—Choline acetyltransferase (ChAc) content of 50 separate rat brain stem nuclei and cerebellum removed by microdissection was determined using a sensitive radiometric assay. The distribution of ChAc activity is uneven, with extremely high levels in the cranial motor nuclei and the nucleus salivatorius. Low ChAc concentrations were observed in the cranial sensory nuclei, the nuclei of the reticular formation, the raphe nuclei and the nuclei of the acoustic system. The lowest ChAc levels were measured in the cerebellum. Comparison of the distribution of ChAc with histochemical localization of acetylcholinesterase revealed generally good agreement, and notable exceptions are discussed.     

CHOLINE ACETYLTRANSFERASE ACTIVITY OF HUMAN BRAIN TISSUE DURING DEVELOPMENT AND AT MATURITY

Abstract— (1) On analysis of human brain tissue to determine its choline acetyltransferase (ChAc) content the recovery of enzyme from many regions is very poor when the tissue is acetone-dried and then extracted in the standard manner; for this reason the method is unsuitable when quantitative recoveries are required; it is preferable to prepare sucrose homogenates and activate these with ether before incubation.
(2) From measurements made on homogenates of one adult brain the highest concentration of ChAc was found in the putamen and the lowest in the corpus callosum. The caudate nucleus also had a high activity. As in other mammals, the concentration of enzyme in the cerebellum was found to be low. Analogous results were obtained on a nine-year-old brain but the level of ChAc activity was generally higher than in the older brain.
(3) During foetal development up to thirty-two weeks, ChAc is higher in the cerebellum than in the caudate, the thalamus, corpora quadrigemina, medulla and spinal cord. In all regions the concentration and total amount of enzyme rise fairly steadily up to this time; between 24 and 32 weeks, however, its concentration in the cerebellum and corpora quadrigemina falls slightly although the total increases considerably.
(4) Comparison of the results with the data of other authors indicates general agreement between the distribution of the enzyme in the human brain and its distribution in other mammals, especially the rhesus monkey. The corpus callosum may be an exception since in man it contains little ChAc while in lower mammals it seems to have relatively high concentrations of both ACh and ChAc.
(5) In comparing the values for ChAc reported here with the values for AChE reported by others, three tissues, the globus pallidus, substantia nigra and cerebellum are found to be exceptional in that relative to their concentration in the caudate the activity of ChAc is only about one-tenth that of AChE.     

THE EFFECTS OF BOTULINUM TOXIN ON ACETYLCHOLINE METABOLISM IN MOUSE BRAIN SLICES AND SYNAPTOSOMES

The effects of Type A botulinum toxin on acetylcholine metabolism were studied using mouse brain slice and synaptosome preparations. Brain slices that had been incubated with the toxin for 2h exhibited a decreased release of acetylcholine into high K⁺ media. Botulinum toxin did not affect acetylcholine efflux from slices in normal K⁺ media. When labeled choline was present during the release incubation, a‘newly-synthesized’pool of acetylcholine was formed in the tissue. In toxin-treated slices exposed to high K⁺, both the production and the release of this‘newly-synthesized’acetylcholine were depressed. A possible explanation for these actions of botulinum toxin would be via an inhibition of the high affinity uptake of choline. This hypothesis was tested by measuring the high affinity uptake of [³H]choline into synaptosomes prepared from brain slices. Previous exposure of slices to botulinum toxin caused a significant reduction in the accumulation of label by the synaptosomes. These data are discussed in terms of our current understanding of the mechanism of action of botulinum toxin and the toxin's interaction with the mechanisms regulating acetylcholine turnover.     

微波对小白鼠超氧化物歧化酶和脂质过氧化物的影响

低剂量（９１５兆赫）的微波照射小白鼠后，用ＮＢＴ光化还原法测知脾脏的ＳＯＤ活性降低；聚丙烯酰胺凝胶电泳ＰＡＧＥ测知ＳＯＤ的电泳图谱发生了变化；脂质过氧化物增加。经恢复饲养，ＳＯＤ的活性持续下降，脂质过氧化物含量持续增高，但降低和增加的趋势于第１１天后趋于稳定，而ＳＯＤ的电泳酶谱却不能恢复到对照水平。本研究表明低剂量的微波照射对小白鼠的ＳＯＤ具有明显的损伤作用，并使有害于机体的脂质过氧化物增加。     

EFFECT OF PRETREATMENT UNDER VARIOUS CATIONIC CONDITIONS ON ACETYLCHOLINE CONTENT AND CHOLINE TRANSPORT IN RAT WHOLE BRAIN SYNAPTOSOMES

The effects of different ionic environments were measured on the concentration of acetyl-choline (ACh) from synaptosomes and their effect on subsequent high affinity choline (Ch) transport and ACh synthesis after resuspension of the synaptosomes in the normal Krebs medium. KCl (40 mM) was used to induce ACh release and reduce synaptosomal ACh content. The effects of Na⁺ omission, Ca²⁺ omission, and high Mg²⁺ on spontaneous (KC1: 4.75 mM) and potassium induced (KC1: 40 mM) ACh release and other cholinergic parameters are presented. The high affinity transport of Ch was more highly correlated with the reciprocal of the ACh level (r= 0.934, P= 9.7 × 10^-4) than with the ACh release rate during preincubation (r= 0.792, P= 3.4 × 10^-2). The results are consistent with the view that the consequences of the various ionic conditions on Ch transport and ACh synthesis are dependent on their effects on intrasynaptosomal ACh levels and only secondarily on synaptosomal ACh release.     

成年与老年大鼠耳蜗核胆碱乙酰化酶免疫组织化学研究

采用免疫组织化学Ｓ－Ｐ法,对成年（２～３月龄）和老年（２０～３６月龄）Ｗｉｓｔａｒ大鼠耳蜗核组织进行ＣｈＡＴ免疫组织化学显色,并利用计算机图像分析系统对免疫反应阳性细胞进行计数。观察老年大鼠耳蜗核组织胆碱乙酰化酶（ＣｈＡＴ）免疫反应的变化,研究其与老年性聋的发病关系。结果发现ＣｈＡＴ免疫反应阳性神经元主要散在分布于背侧核（ＤＣＮ）的梭型细胞层,老年大鼠耳蜗背侧核ＣｈＡＴ阳性细胞数目少于成年组（Ｐ＜０．００１）。实验结果提示,耳蜗核ＣｈＡＴ阳性细胞数目的减少可能与老年性聋的发病有关。     

在胚胎发育中第三脑室室管膜细胞表达乙酰胆碱转移酶

目的在研究胚胎14天（E14）大鼠端脑放射状胶质细胞与胆碱能前体细胞的关系时发现,第三脑室室管膜细胞和侧脑室脉络丛上皮细胞均表达胆碱能神经元标志性蛋白,即乙酰胆碱转移酶（Choline acetyltransferase,ChAT）。方法E14大鼠端脑冠状连续切片,采用免疫组织化学染色法,分别对放射状胶质细胞标志性蛋白,即波形蛋白（Vi men-tin）和乙酰胆碱转移酶进行免疫组织化学染色。结果E14大鼠端脑室管膜细胞呈单层衬于第三脑室,在背中侧的室管膜细胞由单层增殖为复层。上皮基底部与邻近组织分界不清,这些细胞的游离面胞质呈乙酰胆碱转移酶免疫反应阳性,在邻近的第三脑室周围局部形成乙酰胆碱转移酶阳性细胞群。在相邻切片的同一区域内复层室管膜细胞波形蛋白免疫反应阴性。结论第三脑室背中侧室管膜细胞作为神经干细胞分裂、增殖,产生胆碱能前体细胞,并向第三脑室周围组织迁移,形成基底前脑神经核团的Ch5,Ch6胆碱能神经细胞群。