Metabolic profile analysis of free amino acids in experimental autoimmune uveoretinitis rat plasma

AIM: To determine the differences of amino acid(AA) levels in experimental autoimmune uveoretinitis(EAU). METHODS: AA analysis of the plasma samples in EAU rats induced by interphotoreceptor retinoid-binding protein emulsion were performed with high performance liquid chromatography(HPLC) and phenylisothiocyanate(PITC) pre-column derivation methods were performed. Using partial least squares discriminant analysis(PLS-DA), the potential biomarkers were identified in EAU rat plasma, and the metabolic pathways related to EAU were further analyzed. RESULTS: The method results showed that linear(r≥0.9957), intra-day reproducible [relative standard deviation(RSD)=0.04%-1.33%], inter-day reproducible(RSD=0.06%-2.07%), repeatability(RSD=0.03%-0.89%), stability(RSD=0.05%-2.48%) and recovery(RSD=1.98%-4.39%), with detection limits of 0.853-11.4 ng/mL. The metabolic profile in EAU rats was different from that in the control groups five AAs concentrations were increased and nine AAs were reduced. Moreover, five metabolic pathways were related to the development of EAU. CONCLUSION: The developed method is a simple, rapid and convenient for determination of AAs in EAU rat plasma, and these findings will provide a comprehensiveinsight on the metabolic profiling of the pathological changes in EAU.

Effect of nitric oxide synthase inhibitor N^G-nitro-L-arginine on the content of amino acid in ischemic brain tissues of rats

BACKGROUND： Nitric oxide synthase （NOS） inhibrtors have been widely used to investigate the role of NO on cerebral ischemic injury, but the results are controversial. Moreover, it has been considered to aggravate the ischemic neuronal damage with the release of excessively excitatory amino acids （EAA） during cerebral ischemia. On the other hand, some inhibitory amino acid is suggested to be important for the neuronal protection against ischemic brain damage. Our study has recently showed that treatment with the NOS inhibitor NG-nitro-L-arginine （L-NA） reduced focal cerebral ischemic damage. The effect of L-NA on the contents of excitatory and inhibitory amino acid in the rat brain following cerebral ischemia is still unclear. OBJECTIVE： By evaluating the effect of NOS inhibitor, L-NA on the contents of aspartate, glutamate, glycine and γ-aminobutyric acid （GABA） in striatum, hippocampus and cortex in the rat brain following cerebral ischemia respectively, to investigate the beneficial effect of L-NA on cerebral ischemic injury and the possible mechanism. DESIGN： A randomized and controlled experiment SETTING ： Department of Pharmacology, Hebei Academy of Medical Sciences MATERIALS： A total of 42 male healthy SD rats （grade Ⅱ, weighting 250-300 g） were provided by the Experimental Animal Center of Hebei Province （Certification： 04036）. Aspartate, glutamate, glycine, GABA, L-NA and 2,3,5-triphenyltetrazolium chloride （TTC） were obtained from Sigma Chemicals Co, St Louis, MO, USA. HPLC-ultraviolet detector system consisted of Agilent 1100 HPLC. METHODS： The experiment was carried out in Department of Pharmrcology, Hebei Academy of Medical Sciences from June 2005 to June 2006. Rats were randomly divided into three groups： sham-operated group （n = 6）, ischemic group （n = 18）, L-NA group （n = 18）. The model of focal cerebral ischemia in rat was prepared with intraluminal line occlusion methods. In sham-operated rats, the external carotid artery was surgically prepared, but the filament was not inserted. Each group was further divided into 3 subgroups （n = 6 for each）： drugs were administrated at 2, 6 and 12 hours after the middle cerebral artery occlusion （MCAO） respectively. L-NA （20 mg/kg, ip） was administrated, twice a day, for 3 consecutive days. Same volume of normal saline was administrated in ischemic and sham operation groups. The changes of infarcted volume and the contents of amino acids were respectively assayed. Image analysis software was used for the measurement of the infarcted area. The results were expressed as a percentage of the infarcted volume of cerebral/volume of whole brain （IV%） in order to control for edema formation. The contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in the rat brain following cerebral ischemia were respectively measured by HPLC method. All data were analyzed with one-way ANOVA and Dunnett＇s test. MAIN OUTCOME MEASURES： （1） The volume of cerebral infarction; （2） The contents of aspartate, glutamate glycine and GABA in brain tissue after cerebral ischemia. RESULTS ： All 42 rats were involved in the final analysis. （1） Infarcted volume： Volume was 0 in sham-operated group. When L-NA was administrated at 2 and 6 hours after MCAO, the infarcted volume was （20.13±3.59）% and （23.12±5.84）% in L-NA group, which was not similar to that in ischemic group [（22.10±3.98）%, （25.38± 5.37）%, P〉 0.05]. However, the infarcted volume was markedly decreased compared with that of ischemic group when L-NA was administrated at 12 hours after MCAO [（26.11±3.55）% and （37.15±3.58）%, P 〈 0.01]. Changes of amino acid content： At 2 and 6 hours after ischemia, the contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in ischemic group were significantly increased compared with those in sham-operated group （ P〈 0.05-0.01）. However, contents in L-NA group were similar to those in ischemic group （P 〉 0.05）. At 12 hours after ischemia, the contents of aspartate [（0.21 ±0.06）, （0.36±0.05）, （0.29±0.12） mg/g] and glutamate [（0.55±0.06）, （0.78±0.10）, （0.52±0.10） mg/g] in striatum, hippocampus and cortex in L-NA group were significantly decreased compared with those in ischemic group [（0.49±0.17）, （0.63± 0.03）, （0.51±0.15） mg/g; （0.98±0.30）, （1.15±0.15）, （0.93±0.15） mg/g, P〈 0.05-0.01]. Glycine in hippocampus was （0.40±0.07） mg/g, which was higher than that in ischemic group [（0.21±0.07） mg/g, P 〈 0.05]. GABA in striatum, hippocampus and cortex was （0.93±0.10）, （0.62±0.12） and （0.81 ±0.10） mg/g, respectively, which was higher than that in ischemic group [（0.60±0.08）, （0.37±0.17）, （0.59±0.10） mg/g, P 〈 0.05-0.01]. CONCLUSION ： It may be concluded that L-NA have beneficial effect on ischemic cerebral injury in ischemic later stage in rats. The possible mechanism is that L-NA can decrease the contents of aspartate and glutamate, increase the contents of glycine and GABA.

Effect of topiramate on partial excitatory amino acids in hippocampal dentate gyrus of rats after alcohol withdrawal

BACKGROUND： Many researches have indicated that the imbalances of various amino acid transmitters and neurotransmitters in brain are involved in the formation of alcohol withdrawal, especially that glutamic acid is one of the important transmitters for alcohol tolerance in central nervous system. OBJECTIVE： To observe the changes of excitatory amino acids in hippocampal dentate gyrus in rats with long-term alcohol drinking after withdrawal under consciousness, and investigate the therapeutic effect of topiramate on alcohol withdrawal. DESIGN ： A randomized control animal experiment SETTING ： Department of Neurology, Affiliated Hospital of Yanbian University MATERIALS： Thirty male Wistar rats of 4 months old, weighing 300-350 g, were purchased from the Experimental Animal Department, Medical College of Yanbian University. Topiramate was produced by Swish Cilag Company, and the batch number was 02CS063. METHODS： The experiments were carried out in the Department of Physiology, Medical College of Yanbian University from August 2005 to February 2006. ① The rats were divided randomly into three groups： control group （n=10）, alcohol group （n=10） and topiramate-treated group （n=10）. Rats in the alcohol group and topiramate-treated group were given intragastric perfusion of 500 g/L alcohol （10 mL/kg）, once a day for 4 weeks successively, and then those in the topiramate-treated group were treated with 80 mg/kg topiramate at 24 hours after the last perfusion of alcohol, once a day for 3 days successively. Rats in the control group were intragastricly given isovolume saline. ② The withdrawal symptoms were assessed at 6, 30, 48 and 72 hours after the last perfusion of alcohol by using the withdrawal rating scale set by Erden et al, which had four observational indexes of stereotyped behaviors, agitation, tail stiffness and abnormal posture, each index was scored by 5 points, the higher the score, the more obvious the symptoms. ③ The contents of aspartic acid and glutamic acid in hippocampal dentate gyrus were detected with microdialysis technique and high-performance liquid chromatograpy （HPLC） respectively at 6, 30, 48 and 72 hours after the last perfusion of alcohol in the three groups. MAIN OUTCOME MEASURES ： ① Scoring results of alcohol withdrawal symptoms; ② Changes of the contents of aspartic acid and glutamic acid in hippocampal dentate gyrus at the alcohol withdrawal symptoms, and the effects of topiramate. RESULTS： Seven rats were excluded due to inaccurate localization and natural death, and 23 rats were involved in the analysis of results. ①In the alcohol group, the scores of alcohol withdrawal symptoms at 30 and 48 hours after the last perfusion of alcohol were obviously higher than those in the control group （10.50±0.96, 14.17±1.25; 3.50±0.92, 3.16±0,31; P 〈 0.01）. In the topiramate-treated group, the scores at 30 hours after the last perfusion of alcohol （6.06±0.82, 3.50±0.92, P 〈 0.05）, and the withdrawal scores at 48 and 72 hours were close to those in the control group （4.57±0.58, 3.30±0.71; 3.16±0.31, 3.66±0.67; P 〉 0.05）.② Changes of the contents of glutamic acid in hippocampal dentate gyrus： In the alcohol group, the content of glutamic acid at 48 hours after the last perfusion of alcohol was significantly increased as compared with that at 6 hours [（143.32±11.42）%, （99.12±0.69）%; P 〈 0.05], and that at 72 hours was close to that at 6 hours [（78.50±16.40）%, （99.12±0.69）%; P 〉 0.05]. The contents of glutamic acid had no obvious differences at 6, 30, 48 and 72 hours after the last perfusion of alcohol in the topiramate-treated group [（100.30±0.37）%, （118.91±10.40）%, （99.55±12.81）%, （99.08±11.42）%; P 〉 0.05], The content of glutamic acid at 48 hours after the last perfusion of alcohol in the topiramate-treated group was obviously lower than that in the alcohol group （P 〈 0.05）, and those at 30 and 72 hours were close （P 〉 0.05）. ③ Changes of the contents of aspartic acid in hippocampal dentate gyrus： In the alcohol group, the contents of aspartic acid at 30 and 48 hours after the last perfusion of alcohol were significantly increased as compared with that at 6 hours [（126.60±8.67）%, （129.17±10.40）%, （99.25±0.87）%; P 〈 0.05], and that at 72 hours was close to that at 6 hours [（89.87±9.93）%, （99.25±0.87）%; P 〉 0.05]. The contents of aspartic acid had no obvious differences at 6, 30, 48 and 72 hours after the last perfusion of alcohol in the topiramate-treated group [（100.27±0.32）%, （120.81 ±12.63）%, （98.91±7.83）%, （85.92±8.07）%; P 〉 0.05]. The content of aspartic acid at 48 hours after the last perfusion of alcohol in the topiramate-treated group was obviously lower than that in the alcohol group （P 〈 0.05）, and those at 30 and 72 hours were close （P 〉 0.05）. CONCLUSION： ① The occurrences of alcohol withdrawal symptoms are correlated with the increased contents of excitatory amino acids in hippocampal dentate gyrus in rats. ② Topiramate can alleviate the alcohol withdrawal symptoms, which may be correlated with the decreased contents of excitatory amino acids in hippocampal dentate gyrus in rats.

Dysfunction of branded-chain amino acids catabolism in rat cardiac allograft

Objective Allograft vasculopathy ( AV) ,feature of chronic rejection,is a major serious long - term post - operation complication in organ transplantation. The accurate mechanisms for AV have not been definitively established,but extensive basic and clinical studies dem-

Alteration of Excitatory Amino Acid in Experimen-tal Spinal Cord Injury in Rats

Objective To detect the effect of excitatory ammo add (EAA) in the secondary damage following spinal cord injury (SCI). Methods Glutamate (Glu) and Aspartate (Asp) on the injury site (T8) were studied using a rat SCI model induced by Allen's weight drop method (10g×2. 5cm). The result suggested that Asp and Glu were significantly increased in 10 mm. Results Glu was significantly decreased from 2 h to 24 h,while Asp was a tittle reduced in 2 h,and slightly rose in 4 h as compared with Control Group. Though elevated in 8 h,it dropped again in 24h as compared with Control Group. Conclusion The result indicates that the rise of EAA following SCI could be the cause of the secondary spinal cord damage.

Dietary <i>γ</i>-Aminobutyric Acid Shortens the Life Span of Stroke-Prone Spontaneously Hypertensive Rats

Dietary γ-amino butyric acid (GABA) has been suggested to decrease systolic blood pressure. This study aimed to ex-amine the effects of dietary GABA on the life span of stroke-prone spontaneously hypertensive rats (SHRSPs). In this study, life span was determined for SHRSPs provided 1% NaCl solution or 0.01% GABA in 1% NaCl solution as drinking water. The life span of the GABA-fed group (76.3 ± 1.65 days) was significantly shorter than that of the control group (81.6 ± 0.88 days). The results of this study may not be applicable to humans. Future studies will be necessary to elucidate the mechanism underlying this phenomenon.

Synergistic renoprotective effect of a compiled branched-chain amino acids and Cymbopogon schoenanthus extract against experimentally induced oxido-nitrosative renal insult

Objective: To better investigate the protective role of branched-chain amino acids(BCAAs)and Cymbopogon schoenanthus(CS) extract against the potassium dichromate(PDC)-induced oxido-nitrosative nephrotoxic insult in the experimental rat model. Methods: Thirty male rats were randomly divided into five equal groups: The 1 st group served as control; the 2^(nd)was injected with a single dose of PDC(15 mg/kg b.w i.p.);the 3^(rd), 4^(th), and 5^(th) groups were respectively treated with BCAAs, CS, and their combination for 15 d prior to induction of renal insult via PDC single dose(15 mg/kg b.w s.c.). The experimental period was terminated in all groups 2 d after induction of renal insult. The harvested kdney samples were divided for biochemical assays and histological examination. Results: The PDC-induced nephrotoxic effect caused a depletion of renal oxidative scavengers glutathione, superoxide dismutase with consequent lipo-oxidative cellular membrane deterioration manifested by a rise in malonaldehyde, oxidized glutathione, myeloperoxidase and the concomitant increase in inflammatory response elements tumor necrosis factor α, nitric oxide, and interleukin 1 β.Moreover, the comet assay and increased 8-hydroxy-2-deoxyguanosine proved an accelerated apoptotic DNA fragmentation. These local renal changes were met with global altered blood biochemistry. The BCAAs and CS or their compiled administration showed an ameliorative effect against PDC-induced nephrotoxic in a synergistic pattern. Conclusions: Both BCAAs and CS or their combined administration afford potential competitors against renal insult induced by polyvalent anion pollutants in experimentally studied animals model. As a route for novel drug discovery, further investigation should be attempted to optimize their augmenting reno-protecting potential.

安宫牛黄丸对大鼠自发性脑出血模型血肿周围脑组织氨基酸表达及超微结构的影响研究

目的:自发性脑内出血发病率高,现代医学不论是外科治疗还是内科治疗效果均不理想。由于血肿形成后的原发性组织损害已无法恢复,继发性的脑损害预防才是治疗的关键。近期对继发性脑损害的研究重点集中在:血肿周围脑组织血流及代谢改变及其调控因素,继发性脑水肿的发生机制,早期血肿扩大的原理和预防,铁离子超载理论的临床证实及治疗,兴奋性氨基酸理论等方面,并取得了很大的进展。但关于脑保护的实验室工作和临床实践不多,应用于临床的成熟技术、方法更少。

中药复方“体复康”对运动大鼠血清游离氨基酸影响的实验研究

本实验观察了经强化训练后的大鼠，在一次急性运动后的不同恢复时相血清中总氨基酸、支链氨基酸、丙氨酸、酪氨酸浓度及ＡＡＡ／ＢＣＡＡ比值的变化，以及“体复康软胶囊”的调节作用。结果表明，该药对运动大鼠的氨基酸代谢有积极的调整作用，对大鼠在运动过程中能量的动员和贮备，以及大鼠对运动刺激的适应和运动后的恢复起到重要的促进作用。

不同脑温状态地西泮对大鼠脑缺血组织氨基酸含量的影响

目的探讨不同脑温状态下,地西泮对大鼠脑缺血组织谷氨酸(Glu)、天门冬氨酸(Asp)、甘氨酸(Gly)及γ-氨基丁酸(GABA)含量的影响。方法建立大鼠大脑中动脉缺血再灌注模型,诱导目标脑温,用HPLC荧光法检测轻度高温、常温、亚低温各组脑缺血组织Glu、Asp、Gly及GABA含量。结果(1)与常温假手术组比较,常温脑缺血对照组及常温地西泮组Glu、Asp、Gly显著增高(P均<0.001),GABA在常温脑缺血对照组差异无统计学意义(P>0.05),在常温地西泮组则显著增高(P<0.01)。(2)与常温脑缺血对照组比较,常温地西泮组Glu、Asp、Gly稍降低,GABA稍增高,但差异均无统计学意义(P均>0.05)。(3)与常温地西泮组比较,轻度高温地西泮组Glu、Asp、Gly显著增高(P均<0.001),GABA差异无统计学意义(P>0.05);亚低温地西泮组Glu、Asp、Gly显著降低(P均<0.01),GABA显著增高(P<0.001)。结论亚低温状态下,地西泮显著上调GABA水平,从而有利于地西泮"抑制性保护"机制的建立,可能使地西泮的神经保护作用得到增强。

大鼠血浆中痕量氨基酸的柱前衍生-高效液相色谱串联质谱测定

以1,2-苯并-3,4-二氢咔唑-9-乙基氯甲酸酯(BCEOC)作为柱前荧光衍生试剂,在Hypersil BDS C18(4.6mm×200mm,5μm)反相色谱柱上,荧光检测波长为390nm(激发波长为333nm),采用梯度洗脱实现了20种氨基酸标准品衍生物的分离检测。20种组分的线性范围为51.6fmol^105.6pmol,线性回归系数均大于0.9995;检出限为6.3~177.6fmol(S/N=3∶1)。经柱后串联质谱电喷雾电离源(ESISource)实现了各组分的质谱鉴定,并以酪氨酸(Tyr)衍生物为例进行了质谱裂解机理解析。对A、B、C三组(A:安静对照组;B:运动力竭即刻组;C:力竭恢复12h组)24只大鼠血浆中氨基酸的定量测定结果表明,运动力竭即刻较安静状态大鼠血浆氨基酸含量明显增加;力竭恢复12h后血浆氨基酸水平基本恢复到运动前状态。方法的灵敏度高、重现性好,为大鼠血浆中氨基酸的测定提供了一种新方法。

RP-HPLC法检测大鼠海马中4种氨基酸类神经递质含量的研究

建立了RP-HPLC测定大鼠海马中4种氨基酸类神经递质含量的方法。以2,4-二硝基氟苯（DNFB）为衍生化试剂进行柱前衍生,色谱条件为Diamonsil C18（4.6mm×200mm,5μm）色谱柱,乙腈和醋酸钠缓冲液为流动相,采用二元梯度洗脱,UV350nm检测。结果显示：在18min内,4种氨基酸分离完全,在0.01-100μg/mL范围内线性关系良好,天冬氨酸（Asp）、谷氨酸（Glu）、甘氨酸（Gly）、γ-氨基丁酸（GABA）相关系数分别为0.9985、0.9976、0.9943、0.9924;日内变异系数分别为2.4%、3.8%、3.2%和3.5%;标准加入平均回收率分别为94.80%、93.93%、92.69%和96.74%。

丙酮酸肌酸对大鼠胴体组成及机体氨基酸谱的影响

以大鼠为实验对象,灌胃丙酮酸肌酸(CrPyr),通过观测其对机体胴体组成及其血清与组织中氨基酸谱的影响,探讨丙酮酸肌酸对机体中营养物质的代谢流向及其胴体品质的影响。实验选用40只雄性清洁级SD大鼠,随机分为对照组(C)、低剂量组(L)、中剂量组(M)和高剂量组(H)4个处理组,每个处理组按体质量分别灌胃0、750、1500、3000mg/(kgbw·d)的CrPyr。实验为期7周,第49天宰杀大鼠,借助反相高效液相色谱(RP-HPLC)方法测定血清、组织中氨基酸含量的变化。结果表明:与对照组相比,在不影响饲料转化率的情况下,灌胃CrPyr可显著(P<0.05)地提高其腿肌的相对质量;灌胃CrPyr可显著(P<0.05)或极显著(P<0.01)升高大鼠血清和骨骼肌中主要生糖氨基酸的含量,但对肝脏组织的氨基酸谱则没有明显影响。提示,灌胃CrPyr可能通过升高生糖氨基酸的含量,从而促进大鼠体蛋白的合成。

大鼠服用朱砂后脑内氨基酸类递质含量的变化

研究了朱砂对大鼠脑组织中氨基酸类神经递质含量的影响.将32只Wistar大鼠随机分为低、中、高剂量组和对照组,朱砂灌胃给药14天后,采用高效液相色谱法测定大鼠脑组织中谷氨酸(Glu)、天门冬氨酸(Asp)、甘氨酸(Gly)、γ-氨基丁酸(GABA)和牛磺酸(Tau)的含量,并计算兴奋毒性指数(EI)的变化.与对照组相比较,脑组织中氨基酸类神经递质含量均呈下降趋势,其中Asp和Gly的中、高剂量组差异有统计学意义(P<0.05),Tau和GABA的高剂量组有统计学差异(P<0.05),Glu和EI所有剂量组均有统计学差异(P<0.05).朱砂对氨基酸类神经递质具有一定的抑制作用.

泻肝安神法对睡眠剥夺大鼠氨基酸类神经递质影响的研究

目的探讨泻肝安神法对睡眠剥夺大鼠脑干、海马、下丘脑脑区氨基酸类神经递质含量的干预效应。方法将60只SD大鼠随机分为4组,分别为正常对照组、失眠模型组、中药组、西药组,每组各15只,除正常对照组外的其余三组大鼠一律采用水平台法反复剥夺睡眠7次,每次持续24小时。造模结束后,中药组大鼠灌胃给药泻肝安神汤,西药组灌胃给药地西泮溶解液,失眠模型组灌胃给予纯净水,干预7天后取材,采用高效液相色谱法检测各组大鼠脑干、海马、下丘脑脑区6种氨基酸类神经递质的含量。结果经方差分析发现大鼠剥夺睡眠后脑干氨基酸类神经递质门冬氨酸下降,γ-氨基丁酸升高,下丘脑γ-氨基丁酸下降(P<0.05),而在海马和下丘脑区域各氨基酸类神经递质含量变化不显著(P>0.05)。采取泻肝安神法治疗后大鼠脑干门冬氨酸含量显著升高,下丘脑γ-氨基丁酸含量升高(P<0.05);而采用地西泮片治疗后大鼠脑干区门冬氨酸升高,下丘脑门冬氨酸、γ-氨基丁酸、甘氨酸、牛磺酸升高(P<0.05)。结论泻肝安神法可调节睡眠剥夺大鼠的氨基酸类神经递质,在下丘脑的作用机制类似于地西泮片,但对于氨基酸类神经递质的影响影响靶点要少于西药。

衰老对大鼠脑区氨基酸水平的影响

本文测定了正常青龄组（３月龄）和老龄组（２０月龄）大鼠不同脑区（皮层、小脑海马、纹状体和下丘脑）谷氨酸、天门冬氨酸、甘氨酸、ｒ－氨基丁酸和牛磺酸的含量。结果表明：在衰老过程中大鼠某些脑区谷氨酸、天门冬氨酸、甘氨酸和牛磺酸水平显著降低；而纹状体γ－氨基丁酸含量则显著升高。

复合花粉制剂改善CCl_4诱导肝硬化大鼠血浆氨基酸失衡

目的观察复合花粉制剂对CCl4诱导的肝硬化模型大鼠的血浆氨基酸浓度的影响。方法将45只SD大鼠采用抽签法随机分为模型组(35只)和对照组(10只)。对照组与模型组动物给予同样的基础饮食;模型组按照每100 g体质量0.2 mL的剂量,腹腔注射40%CCl4葵花籽油溶液,每周2次,持续12周,待病理证实肝硬化形成,停止注射CCl4。12周时32只大鼠造模成功,对照组大鼠全部存活,营养素干预前处死模型组大鼠4只,对照组大鼠3只,检测基线血浆氨基酸浓度;余7只对照大鼠分别于营养素干预2周、4周后处死;余28只模型组大鼠随机分为组1(模型对照组)、组2(复合花粉营养素)、组3(高剂量Noveliver营养素)、组4(低剂量Noveliver营养素),分别于营养素干预2、4周后处死,检测各组干预后2、4周血浆氨基酸浓度。结果①基线血浆氨基酸浓度:模型组血浆支链氨基酸及芳香族氨基酸浓度均低于对照组,但2组支链/芳香族氨基酸比值差异无统计学意义(分别为1.48±0.05和1.48±0.13)。②营养素干预2周后血浆支链氨基酸:组3高于组4和组2;芳香族氨基酸:组2和组3低于其余3组;支链/芳香族氨基酸比值:对照组、组1、2、3、4分别为1.54±0.11、1.59±0.43、1.82±0.11、1.98±0.55和1.58±0.04,组3和组2明显高于其余3组。③营养素干预4周后血浆支链氨基酸:3组营养干预组(组2、3、4)均高于模型对照组(组1),其中组3高于组4;芳香族氨基酸:组2和组4低于其余3组;支链/芳香族氨基酸比值:正常对照组、组1、2、3、4分别为1.44±0.14、1.16±0.35、1.40±0.24、1.51±0.08和1.40±0.15,3组营养干预组(组2、3、4)均高于模型对照组(组1),其中组3高于组4。结论 CCl4所致肝硬化大鼠血浆氨基酸整体水平低于正常对照组大鼠;复合花粉制剂营养素干预有提高血浆支链氨基酸水平和支链/芳香族氨基酸比值的趋势,且高剂量Noveliver效果优于低剂量Noveliver,干预4周时效果优于2周时的结果。本实验提示复合花粉制剂有一定改善肝硬化大鼠血浆氨基酸失衡的作用。本研究为临床肝硬化患者氨基酸失衡的治疗提供了参考。

穴位埋线对癫痫大鼠海马神经元细胞凋亡及氨基酸的影响

目的探讨穴位埋线对癫痫大鼠海马神经元氨基酸及细胞凋亡的影响。方法选取50只Wistar大鼠,每组10只,随机分为空白对照组、模型组、穴位埋线组、针刺组、丙戊酸钠组。除空白对照组外,其他4组均用青霉素钠腹腔注射造模,穴位埋线组第1、4、7天重复埋线。第7天模型组、穴位埋线组、针刺组及丙戊酸钠组再一次腹腔注射青霉素钠点燃模型大鼠,所有动物均在第7天治疗后断头取脑,剥离海马。标本切片观察各组大鼠海马神经元形态学改变(原位末端标记法)。并检测海马内谷氨酸(Glu)、?-氨基丁酸(GABA)的含量(高效液相色谱仪),计算GABA/Glu比值。结果空白对照组大鼠海马神经元细胞正常,阳性凋亡细胞稀疏,模型组、针刺组、穴位埋线组及丙戊酸钠组均可见不同程度海马细胞凋亡现象。与模型组相比,穴位埋线组、针刺组及丙戊酸钠组海马细胞凋亡现象明显降低(P<0.01),丙戊酸钠组低于穴位埋线组(P<0.05),穴位埋线组低于针刺组(P<0.05)。模型组海马区兴奋性氨基酸Glu的含量较空白对照组明显增加(P<0.01),穴位埋线组、针刺组及丙戊酸钠组与模型组对比海马内Glu的含量明显减少(P<0.01),且穴位埋线组及丙戊酸钠组Glu水平少于针刺组(P<0.05),穴位埋线组及丙戊酸钠组间无明显差异(P>0.05)。模型组海马区GABA的含量低于空白对照组(P<0.01)。穴位埋线组、针刺组及丙戊酸钠组GABA的含量均高于模型组(P<0.01),穴位埋线组、丙戊酸钠组及针刺组间GABA含量无明显差异(P>0.05)。模型组GABA/Glu比值低于空白对照组(P<0.01),穴位埋线组、针刺组、丙戊酸钠组GABA/Glu比值高于空白对照组(P<0.01),穴位埋线组、丙戊酸钠组及针刺组间GABA/Glu比值无明显差异(P>0.05)。结论重复穴位埋线可明显阻止癫痫大鼠海马神经元细胞凋亡的发生发展并可通过调节脑内兴奋性与抑制性氨基酸的水平而发挥抗癫痫作用。

佐剂关节炎大鼠行为、脑组织氨基酸的变化及新风胶囊对其的影响

目的观察佐剂关节炎(adjuvant arthritis,AA)大鼠行为、脑组织氨基酸的变化及新风胶囊(XFC)对其的影响。方法用弗氏完全佐剂(complete Freund’s adjuvant,CFA)诱发大鼠产生关节炎,设立正常对照组、模型对照组、XFC治疗组、甲氨喋呤(MTX)治疗组、雷公藤多苷片(TPT)治疗组。观察AA大鼠行为、脑组织氨基酸的变化和关节肿胀度、关节炎指数(arthritis index,AI)的变化以及XFC对其的影响,并进行相关性分析。结果与正常对照组比较,模型对照组大鼠自主活动次数明显减少、跳台错误次数均明显增多、大鼠跳台的时间(step down latency,SDL)缩短而反应时间(escape latency,EL)延长,脑组织中γ-氨基丁酸(GABA)含量明显升高,而谷氨酸(GLU)/GABA值明显降低(P<0·05)。与模型对照组比较,XFC组大鼠的自主活动次数显著增加、SDL延长、EL缩短、跳台错误次数减少,GABA明显下调、GLU/GABA上调(P<0·05),而MTX、TPT组改善不明显(P>0·05)。各治疗组大鼠的肿胀度、关节炎指数均明显降低(P<0·05);AA大鼠的自主活动次数、SDL与足跖肿胀度、关节炎指数、甘氨酸(GLY)、GABA呈负相关,与GLU/GABA值呈正相关(P<0·05);EL、跳台错误次数均与关节炎指数、GLY、GABA呈正相关,与GLU/GABA值呈负相关(P<0·05)。结论免疫应激可引起大鼠行为改变及脑组织氨基酸的变化,XFC可通过调节脑组织氨基酸递质改善AA大鼠行为。

雄黄染毒后大鼠脑组织中氨基酸类神经递质含量的变化

研究了雄黄对大鼠脑组织氨基酸类神经递质含量的影响.将32只Wistar大鼠随机分为对照组(0.5%CMC-Na)以及低剂量(0.3g/kg)、中剂量(0.9g/kg)、高剂量(2.7g/kg)雄黄染毒组,通过连续灌胃给予雄黄混悬液两周.采用高效液相色谱-柱前衍生化法测定了大鼠脑组织中氨基酸类神经递质含量的变化.结果表明,与对照组比较,低剂量组大鼠脑组织中丝氨酸、甘氨酸和γ-氨基丁酸含量明显增加.中、高剂量组大鼠脑组织中同型半胱氨酸、谷氨酰胺、丝氨酸和天冬氨酸含量明显比对照组的低.总体而言,雄黄可对大鼠脑组织氨基酸类神经递质产生影响,氨基酸类神经递质可能是雄黄毒性作用的靶点之一.