烫伤后大鼠肝糖皮质激素受体减少的机理——肾上腺部分切除动物的研究

部分去除肾上腺的大鼠烫伤后,血清糖皮质激素无明显升高,而肝胞液糖皮质激素受体(GR)却有升高的趋势,其中烫伤后6 h组和对照组的相差有非常显著的意义。进一步用打点杂交进行的研究表明,部分去腺大鼠烫伤后6h肝GR mRNA也有增加。这些结果提示,完整动物烫伤时肝胞液GR的减少依赖于血中糖皮质激素的升高。     

未去肾上腺大鼠肝胞液糖皮质激素受体的测定及烫伤后的变化

在内源性激素存在的条件下,用交换法测定了大鼠肝胞液的糖皮质激素受体。交换条件为Oc4.5～5小时。在这条件下,[~3H]皮质酮—受体复合物的解离将近90％。向去肾上腺大鼠肝胞内预先加入不同量的皮质酮保温后,再测定[~3H]地塞米松特异结合,其结果表明,胞液皮质酮浓度在6nm以内时,[~3H]地塞米松特异结合容量(Ro)的测定值的减少不超过10％,表面上的解离常数(Kd)的测定值基本上不变。用这方法测定烫伤大鼠肝胞液的Ro和Kd,结果Ro减少。烫伤后1、12、24、36小时和5天的Ro分别为对照组的67.4±15.2、70.6±22.0、79.7±5.8、113.0±24.8和102.9±17.6％(均值±标准差)。烫后1小时Kd增大。对其可能的机理作了讨论。     

烫伤大鼠肝组织糖皮质激素受体的变化及α-MSH、合成肽KPV的调节作用

目的　探讨烫伤所致的病理性应激时大鼠肝胞液糖皮质激素受体的变化及可能的调节机制。方法　以 [3H]地塞米松为配体 ,用放射性配基 受体结合分析法测定了正常对照组、轻度烫伤组和重度烫伤组大鼠肝胞液糖皮质激素受体的结合容量(Receptorcapacity ,R0 )和表观解离常数 (Kd)。采用体内注射TNFα、IL 1 β中和抗体和α 促黑色素细胞刺激激素 (α melanocyte stimulatinghormone ,α MSH)和合成肽KPV(Ac D Lys L Pro D Val)等措施调节其改变。结果　与正常组的R0 [Massactionrobust:(30 7.86± 2 4.2 2 )fmol/mg ;Scatchard :(30 6 .71± 2 7.96 )fmol/mg]相比 ,烫伤后 1 2h ,轻烫组的R0 [MassActionRobust:(2 85 .1 9± 1 6 .6 2 )fmol/mg ;Scatchard :(2 96 .6 4± 1 6 .0 6 )fmol/mg]差异不显著 ;重烫组的R0 [Massactionrobust:(2 0 5 .5 2± 30 .41 )fmol/mg ;Scatchard :(2 0 8.45± 30 .78)fmol/mg]则显著下降 (P <0 .0 5 )。用体内注射TNFα、IL 1 β中和抗体和α MSH和KPV均能明显改善重烫组R0 的降低。结论　TNFα、IL 1 β中和抗体、α MSH及KPV均可在一定程度上防止重度烫伤所致的病理性应激时糖皮质激素受体的减少     

甲亢类阴虚大鼠肝胞液糖皮质激素受体的实验研究

本研究以甲亢大鼠模拟阴虚证模型,测定其肝胞液糖皮质激素受体(GCR)等。实验结果表明,甲亢类阴虚大鼠肝胞液GCR最大结合容量(R。)较正常对照组减少(P<0.01),而GCR平衡解离常数(Kd)则无明显变化(P>0.05),经养阴益气中药治疗后的甲亢大鼠肝胞液GCR的R。回升,接近正常对照组水平(与阴虚组比P<0.05,与对照组比P>0.05)。提示阴虚证病理实质可能与肝胞液GCR数量减少有关,而养阴药则可以提高肝胞液GCR的数量。该实验结果为进一步研究阴虚证与GCR关系提供了有价值的实验依据。实验结果还表明,甲亢类阴虚大鼠的肾上腺皮质分泌功能亢进;养阴益气中药可降低甲亢大鼠的T_8水平及对该动物模型的多种病理改变有不同程度的纠正作用。     

甲状腺激素对大鼠肝糖原及肝胞液糖皮质激素受体的影响

用实验性甲状腺功能亢进大鼠,同时观察甲状腺激素对肝糖原含量的影响和肝胞液糖皮质激素受体(GCR)的变化。结果表明,甲状腺片(75 mg/kg·d,ig×10 d和100 mg/kg·d,ig×10d)所致甲状腺功能亢进,可同时引起与甲状腺片剂量有依赖关系的肝糖原减低和与之平行的肝胞液GCR数目升高。提示,甲状腺激素减低大鼠肝糖原含量的作用与肝胞液GCR的增高有直接关系。     

严重烫伤大鼠肝脏热休克蛋白90α的表达变化及意义

目的 研究严重烫伤早期大鼠肝脏组织中糖皮质激素受体分子伴侣热休克蛋白90α(HSP90α)的表达变化及意义。方法 用免疫印迹技术分析严重烫伤后不同时相肝脏组织中HSP90α的表达量改变，通过免疫组化进一步观察HSP90α在肝细胞胞核中的表达变化。结果 大鼠严重烫伤后72h内肝脏HSP90α的表达明显升高，尤其以12～48h更为显著，在48h左右肝细胞核内还出现HSP90α的异位高表达。结论 严重烫伤可以引起肝脏HSP90α异常高表达，这可能是严重创伤早期糖皮质激素受体表达和功能发生改变的原因之一。     

参附汤对失血性休克大鼠糖皮质激素及其受体的影响

深入探讨参附汤的药理作用机理。以急性失血性休克大鼠为实验对象，同步观察参附汤对血浆皮质酮及肝胞液，胸腺细胞糖皮质激素受体的影响。参附汤组大鼠肝胞液及胸腺细胞ＧＲ的结合位点都明显高于单纯失血组，参附汤组血浆皮质酮略高于失血组。参附汤可纠正失血性休克模型的ＧＲ的减少，以发挥其救治休克，回阳固脱的作用。     

大鼠肝、肺胞液糖皮质激素受体的超离心分析

本文用蔗糖密度梯度超离心技术分析了大鼠肝、肺细胞液糖皮质激素受体复合物,结果为在低离子强度介质中,该受体复合物的沉降行为表现为7～8S;在10mM钼酸钠介质中,肝胞液糖皮质激素受体复合物的沉降行为表现为9～10S。本文还对影响该受体复合物稳定性的一些因素作了讨论。     

严重烧伤后胃黏膜细胞胞液中糖皮质激素受体与胃黏膜电位差的相关性研究

目的了解严重烧伤后胃黏膜细胞胞液中糖皮质激素受体变化与胃黏膜电位差之间的关系.方法观察了大鼠严重烧伤后不同时期胃黏膜细胞胞液糖皮质激素受体的动态变化及胃黏膜电位差变化情况.结果严重烧伤后3 h胃黏膜组织皮质醇含量明显升高,12 h达高峰,而胃黏膜细胞胞浆内糖皮质激素逐受体的最大结合容量(R0)显著下降,平衡解离常数(KD值)明显增大,伤后12,24h胃黏膜电位差亦显著下降.结论糖皮质激素受体水平降低是严重烧伤后急性胃黏膜屏障功能受损的重要原因之一.     

糖皮质激素对兔脑,肺和肝细胞糖皮质激素受体的调节

采用放射配基结合分析法，研究地塞米松对兔脑、肺、肝的糖皮质激素受体的调节作用。结果发现，静脉注射Ｄｅｘ５ｍｇ／ｋｇ后兔肺、肝胞液ＧＲ的结合量明显下降，与对照组比较Ｐ＜０．００１，脑胞液ＧＲ与对照组比较无显著差异。提示糖皮质激素对兔不同靶器官ＧＲ的调节存在不同特点。     

Determination of the Molecular Weight of Photoaffinity Labeled Glucocorticoid Receptor in Scalded Rat Liver Cytosol

Previous work in our laboratory showed that,after scalding,there wasnot only a decrease of rat liver cytosol [~3H]-Dexamethasone specific binding sitesbut also an increase of apparent dissociation constant of [~3H]-Dex and change ofsedimentation coefficient of glucocorticoid receptor.The present study was carriedout using photoaffinity labeling of glucocorticoid receptor in rat liver cytosol.Itwas found that there was no significant difference between molecular weights ofglucocorticoid receptor of normal and scalded rats determined by SDS-polyacrylamide gel electrophoresis.Our result suggests that chnage ofsedimentation coefficient is not due to alteration of molecular weight ofglucocorticoid receptor binding subunit.     

糖皮质激素受体水平在白血病、肾病治疗中临床意义的探讨

目的:探讨糖皮质激素受体(GCR)水平高低对急性淋巴性白血病(ALL)、肾病综合征(NS)疗效的影响。方法:采用受体放射配体结合技术测定ALL、NS和对照组外周血淋巴细胞糖皮质激素高亲和力受体GCRH和低亲和力受体GCRL位点数,分析GCR与疗效之间的关系;并通过动物实验观察中药小柴胡汤对应用糖皮质激素所产生的GCR“降调节”效应的影响。结果:疗前GCRH>4000位点/细胞的ALL患者对激素联合化疗疗效好,化疗后GCRH明显降调,但GCRL治疗前后无明显变化。NS患者疗前GCRH>6000位点/细胞,则对GC疗效好,当GCRH<3000位点/细胞,则GC治疗无效。SD大白鼠实验表明,小柴胡汤具有显著的调控GCR水平作用。结论:对ALL患者和NS患者治疗前检测GCR水平,有助于制定个体化的治疗措施。     

Patterns of adrenoceptor change during liver regeneration of the Wistar Kyoto rat: a binding study.

BACKGROUND: Adrenoceptors have been involved in the regulation of hepatocyte proliferation after partial hepatectomy, as well as in primary culture. This report characterizes alpha 1- and beta-adrenoceptor change during the time-course of liver regeneration in adult Wistar Kyoto rats. METHODS: Saturation binding assays with [3H]prazosin or [3H]dihydroalprenolol (for alpha 1- and beta-adrenoceptors, respectively) were done in liver plasma membranes from 6-month-old rats subjected to 70% hepatectomy followed by hepatic regeneration. RESULTS: [3H]Prazosin and [3H]dihydroalprenolol binding gave control Bmax values of 101 +/- 10 and 12 +/- 1 fmol/mg protein and Kd of 0.50 +/- 0.10 and 4.1 +/- 0.4 nM for alpha 1- and beta-adrenoceptors, respectively. alpha 1-Adrenoceptor number and Kd increased at 24 and 48 h and returned to control values at 72 and 96 h after surgery, whereas beta-adrenoceptors augmented at 48 and 72 h, with a Kd change at 24 and 48 h posthepatectomy. CONCLUSIONS: These results suggest that dual control of alpha 1- and beta-adrenoceptor membrane expression could be involved in different steps during hepatocyte proliferation, and that Wistar Kyoto rats have a different adrenoceptor pattern expression from other rat strains.     

Alteration of Glucocorticoid Receptor Following Forebrain Ischemia in Gerbil

Althoughglucocorticoid(GC)levelsincreaseinbrainischemia,massivedosesofGCarestillusedinbrainischemia,especiallyinbrainedema[1,2].ManyhypothesisconcerningGCagainstbrainischemiahavebeenproposed.Forexample:GCcouldincreasebloodflowinbrainlesionarea,inhibitlipidperoxidereaction,stableionchannelofcellmembrane,improveCa2 outflowfromcell,decreaseCa2 overloadincellandprotectneuronfromsecondarylesionanditisanadaptivereactionofbodythatGCreleaseincreaseinischemia[3].Buttheprofoundmechanismisnotclear.T…     

Regeneration of functionally active rat brain muscarinic receptor in vitro after inhibition with methylmercury chloride]

The effect of methylmercury on muscarinic receptors and the regeneration of functionally active muscarinic receptor in vitro by antagonists were investigated. The result showed that methylmercury chloride (MMC) inhibited the binding of [3H] QNB to muscarinic receptor of rat brain-lysed synaptosomes, with IC50 values of 4.18 mmol/L. Regeneration of functionally active rat brain muscarinic receptors after inhibition with methylmercury was achieved by 2,3-dimercapto-1-propanesulfonc acid, Na salt (DMPS) dithiothreitol (DTT), glutathione (GSH) and Cysteine. Blocking the sulfhydryl groups is suggested to be the molecular mechanism of inhibition of brain muscarinic receptors by methylmercury. Our results provide evidence that thiols chelate out mercuric cations that tightly bound to sulfhydryl groups in muscarinic receptor binding sites and regenerate [3H] QNB binding activity.     

Effect of Amlodipine on the Growth of Vascular SmoothMuscle Cells of Spontaneously Hypertensive Rats

Abnormalvascularsmoothmusclecell(VSMC)proliferationisanimportantpathologicalcharacteristicofhypertension.IthasbeenapopularsubjectthathowtoinhibitVSMCproliferationintheresearchofhypertensioncontrol.Nowwhetheraantihypertensivedrugcanpreventandregresshypertrophyofyascularwallandleftventriclehasbeenbelievedtobeoneofimportantcriteriaintheevaluationofananti-hypertensivedrug.Amlodipineisoneofcalciumantagonistofdihydropyriodines.ItreducesbloodpressurebyblockingcalciumchannelsonVSMCmembrane.Butno…     

大鼠脑突触质膜甘氨酸受体的研究

本文用放射配体受体结合法对大鼠脑突触质膜上甘氨酸受体进行了测定，同时也研究了两型甘氨酸受体在脑内的不同分布。结果显示，甘氨酸受体（士的宁不敏感型）结合容量民为２６±４Ｐｍｏｌ／ｍｇｐｒｏｔｅｉｎ，平衡解离常数Ｋｄ值为１７３±１５ｎＭ，ＩＣ５０值为１０－６Ｍ。除脑干部甘氨酸受体为土的了敏感型外，余脑部主要为土的宁不敏感型。     

The purification of nucleoside diphosphate kinase from rat brain.

Nucleoside diphosphate kinase from the cytosol of rat brain was purified to electrophoretic homogeneity through a series of chromatography columns, including DE-52, AcA-34 Ultrogel, phenyl-Sepharose and Q-Sepharose. The molecular weight of the enzyme was 64,000 as estimated by gel filtration, and it consisted of 3 identical 19,000 subunits. [35S]-GTP gamma S binding and autophosphorylation of the enzyme, and inhibition of the GTPase activity of G-protein were used to screen for the enzyme. These methods were shown to be useful for purifying and assaying the enzyme.     

发育期单眼形觉剥夺弱视模型猫视皮层谷氨酸受体状况的研究

目的 观察单眼形觉剥夺性弱视动物模型谷氨酸受体(GluRs)的变化,探索形觉剥夺性弱视发生的物质基础.方法 11只初生家养猫随机分为两组,处理组在3周龄时缝合单侧眼睑塑造单眼形觉剥夺性弱视模型,3月后用受体的放射性配体结合分析实验观测视皮层GluRs的结合位点数和亲和力.对照组未予特殊处理.结果 处理组视皮层GluRs结合位点数较对照组下降(P＜0.001),而KD值高于对照组(P＜0.001).两组放射性配体-受体结合实验的Hill系数均接近于1.L-[3,4-~3H]-谷氨酸与各型GluRs结合位点的亲和力相同,无正负协同效应.结论 单眼形觉剥夺性弱视动物模型视皮层GluRs亲和力低,GluRs参数出现异常,说明形觉剥夺性弱视的发生可能有一定的物质基础.