首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 171 毫秒
1.
钳夹损伤兔右坐骨神经,于损伤处注射蛇毒NGF400Bu/kg/日,损伤术后1,3,7天和2,3,4,6,8周动态观察脊髓腰段伤侧第Ⅸ板层外侧群的大型运动神经元的AChE活性改变。结果表明术后1,3天实验组(指损伤给药组)和对照组(指损伤对照组)AChE活性均下降(P>005);术后1,2,3周对照组AChE活性明显下降,而实验组AChE活性逐渐趋于恢复(P<001);术后6周实验组AChE活性恢复至正常水平(P<001)。本研究显示蛇毒NGF对坐骨神经损伤后脊髓前角运动神经元AChE活性恢复有促进作用,从而对运动神经元可起一定的保护作用和促进恢复的作用  相似文献   

2.
为探讨八肽胆囊收缩素(CCK-8)和阿片肽相互作用的分子机理,利用抗体免疫沉淀技术研究了CCK-8与NDAP(κ阿片受体激动剂)对大鼠脑(去皮层和小脑)和脊髓背柱组织Fos蛋白的影响。结果表明,0.1μmol/L CCK-8可显著刺激脑和脊髓组织中Fos蛋白增加(分别是对照组的3.8倍和3.6倍)。相同浓度的NDAP对Fos蛋白的生成亦有一定的诱导作用,分别是对照组的2.7倍和2.6倍。CCK-8  相似文献   

3.
以改良Alen氏法造成Wistar大鼠不完全性脊髓损伤,采用神经学功能评分法评定大鼠运动功能,应用定量酶细胞化学方法观察脊髓前角运动神经元内乙酰胆碱酯酶(AChE)和酸性磷酸酶(AcP)活性变化。结果显示:1.脊髓损伤后大鼠运动功能障碍,随后逐渐恢复。2.前角运动神经元内AChE活性减弱、AcP活性增强;随后酶活性呈逐渐恢复,四周时AChE活性基本恢复正常。结果说明:大鼠脊髓不完全性损伤后运动功能变化与前角运动神经元的功能状态具有较强的相关性;前角运动神经元在不完全性脊髓损伤运动功能恢复中起重要作用。  相似文献   

4.
通过计算机模拟比较十种理论上柔性较好的接头在 5′ I L6 T N FΔ融合蛋白中对 I L 6 和 T N FΔ空间结构的影响情况,从中选择了 S A P G T P接头.以 S A P G T P 作为接头的 5′ I L6 S A P G T P T N FΔ和以 P G 为接头的5′ I L6 P G T N FΔ空间结构预测结果相似. D N A 序列分析两种蛋白的接头序列均与设计的一致.5′ I L6 S A P G T P T N FΔ和 5′ I L6 P G T N FΔ蛋白的大肠杆菌表达产物经初步分离、纯化及鉴定后,生物学活性及对高表达 I L 6 受体肿瘤细胞的杀伤作用比较结果显示:在 L929细胞上,前者的生物学活性是后者的 27 倍;在 U937 细胞上,前者对肿瘤细胞的抑制率是后者的13 倍.它们对高表达 I L 6 受体的 U937 细胞杀伤作用分别是同样突变位点的人 T N Fα衍生物的37 和 29 倍.实验表明, S A P G T P作为接头构建的 5′ I L6 S A P G T P T N FΔ融合蛋白优于以 P G 作为接头构建的 5′ I L6 P G T N FΔ融合蛋白.  相似文献   

5.
为探讨八肽胆囊收缩素(CCk-8)和阿片肽相互作用的分子机理,利用抗体免疫沉淀技术研究了CCK-8与NDAP(k阿片受体激动剂)对大鼠脑(去皮层和小脑)和脊髓背柱组织Fos蛋白的影响。结果表明,0.1μmol/LCCK-8可显著刺激脑和脊髓组织中Fos蛋白增加(分别是对照组的3.8倍和3.6倍)。相同浓度的NDAP对Fos蛋白的生成亦有一定的诱导作用,分别是对照组的2.7倍和2.6倍。CCK-8和NDAP共同处理组织,Fos蛋白生成水平相似(脑)或高于(脊髓)CCK~-8单独诱导的水平。结果表明,CCK-8和NDAP均可直接诱导大鼠脑和脊髓组织c-fos的表达,它们对c-fos表达的相互作用在脑和脊髓中呈现不同的模式。  相似文献   

6.
吗啡对福尔马林引起大鼠海马内IL-2RβmRNA表达的影响   总被引:1,自引:0,他引:1  
Wu X  Li HD  Li XC  Ruan HZ  Wang J 《生理学报》1998,50(5):591-594
本实验采用原位杂交法观察足底注射福尔马林(For)痛敏对海马内白细胞介素2受体βmRNA(IL-2RβmRNA)生成的影响及其与吗啡、促肾上腺皮质激素(ACTH)的关系。结果表明:正常大鼠海马有IL-2RβmRNA表达,集中分布于CA1-CA4区神经元、齿状回颗粒细胞。足底注射For后6h双侧海马IL-2RβmRNA表达均增加(P〈0.05),12h达高峰,24h仍高于正常。在6h时,腹腔注射吗啡  相似文献   

7.
高粱细胞质雄性不育系3197A(3A)在常温条件下是不育的(Figs.11&2),经热激(45℃)诱导不同程度地恢复了育性(Figs.13&4),为研究其不育机理提供了线索。热激2h后,3A中即可产生一类线粒体热激蛋白(HSPs)。其中,分子量为70kD的HSP70含量最高,也最为稳定。不过,3A中HSPs的稳定性弱于保持系3197B(3B)(Fig.2,Panels1~4)。放线菌素D抑制HSPs的合成,而氯霉素无此作用(Fig.2,Panels5&6),表明:HSPs是由核基因编码、在细胞质中合成、再跨膜转运到线粒体中的。3A幼穗经热激后,线粒体的总蛋白量猛增了2.7倍(Fig.3),达到3B的水平,育性亦变为可育的。Fig.4表明:HSP70反义链cDNA(R1)能进入到3B花药细胞中,并与靶RNA(HSC70mRNA)结合,而对照、正义链cDNA(D)链无此反应。由此、再增加一个通用保守序列的反义链cDNA(R2)、共两个探针(R1、R2),可以检测到:3A在常温下没有能力合成HSC70mRNA(Fig.5),而在热激条件下,转变为有能力(Fig.6)。启示:3A在热激条件下由不育转变为可育  相似文献   

8.
小麦叶绿体中的CTK结合蛋白属热敏感蛋白,它与6-BA结合的最适温度约为25℃;最适pH为8左右;在其蛋白分子上存在两类CTK结合位点,高亲和力位点与6-BA结合的Kα值为1.1×10-9mol/L,低亲和力位点的Kα值为9×10-7mol/L;激动素、玉米素对6-BA的结合只有部分抑制作用,而ABA、GA3、IAA及腺嘌呤则无竞争作用。CTK结合蛋白分子中的中性氨基酸含量很高,在中性介质中带弱负电。  相似文献   

9.
人为调节荔枝果皮有机自由基及其与果皮变褐的可能关系   总被引:5,自引:0,他引:5  
荔枝果实以10ppm的6-BA、NAA、GA3+NAA或PVP(1%)+NAA处理2min,1℃贮藏8天后再于室温放2天,EPR分析表明果皮的有机自由基比对照低9%-22%,花色素苷含量高5%-46%。离体果皮小圆片同法浸泡1h或16h,有机自由基反而比对照高,O2的捕获剂Tiron和抗氧化剂PG+CA处理皮小圆片1h,有机自由基降低8%-11%,处理16h则自由基高于对照。邻苯二酚和H2O2加速  相似文献   

10.
用荧光标记的受体底物(Gnβ1-2Mα1-6(Gnβ1-2Mα1-3)Mβ1-4Gnβ1-4(Fucα1-6)Gn-PA),结合高效液相层析(HPLC)建立了细胞β-1,4-半乳糖基转移酶的活性检测方法.研究了HL60细胞在体外低血清培养后不同的时间其酶活性的变化,发现12至24h酶活性达到一个高峰,为50.14pmol/min(106cel),此时细胞处在分裂间期,其它各测定时间变化不大.用PMA,RA等细胞诱导分化剂处理HL60细胞株时,发现其活性发生了较明显的变化,PMA诱导的细胞其酶活性在24h变化最大,升高到对照的1.32倍;而RA处理的细胞其酶活性在72h变化最大,升高到对照的2.15倍.  相似文献   

11.
12.
The characteristics of receptors for platelet-activating factor (PAF) on rabbit neutrophils are investigated in this report. The presence of PAF-specific binding to rabbit neutrophils was confirmed using radiolabeled ligand binding assays and a rabbit peritoneal neutrophil membrane preparation. Binding of PAF to the neutrophil membranes was reversible and reached equilibrium within 30 min. Scatchard analysis of PAF-specific binding to the rabbit neutrophil membranes revealed a dissociation constant (Kd) for PAF of 0.41 +/- 0.045 nM and a Bmax of 0.32 +/- 0.11 pmol of PAF receptor/mg of protein. The order of potencies of PAF receptor antagonists to inhibit the binding of 3H-PAF to rabbit peritoneal neutrophil membranes was determined. For the competition assays, 100 micrograms of neutrophil or platelet membrane protein, 0.18 nM 3H-PAF, and varying amounts of PAF antagonist were incubated at room temperature for 1 hr. PAF receptor antagonists tested were ONO-6240, brotizolam, kadsurenone, WEB-2086, L-652-731, BN-52021, CV-3988, triazolam, alprazolam, and verapamil. The orders of potencies of these PAF receptor antagonists were similar for inhibition of 3H-PAF binding to rabbit peritoneal neutrophil and platelet membranes (correlation coefficient, r = 0.97). PAF had a significantly higher affinity for rabbit neutrophil membranes (Kd = 0.41 +/- 0.045 nM), as compared with its affinity for rabbit platelet membranes (Kd = 0.87 +/- 0.092 nM). In addition, sodium was found to inhibit 3H-PAF specific binding to rabbit platelet membranes and not to affect 3H-PAF binding to neutrophil membranes. These data indicate that, although PAF receptors on rabbit platelets and neutrophils exhibit similar orders of potencies of PAF receptor antagonists to inhibit the binding of 3H-PAF, the disparity in Kd of PAF for the receptors and the effect of NaCl on the binding of 3H-PAF reveal subtle differences between the cell types.  相似文献   

13.
Steroid binding sites with high affinity for progesterone (Kd=40+/-14 nM determined by binding, and Kd=71+/-22 nM determined by displacement studies) and lower affinity for 21-hydroxyprogesterone and for testosterone, but no affinity for estradiol-17beta, onapristone and alpha-naphthoflavone were detected in the enriched plasma membrane fraction of the fungus Rhizopus nigricans. The amount of steroid binding sites is in accordance with the value of B(max)=744+/-151 fmol (mg protein)(-1). In the membrane fraction, progesterone induced about 30% activation of G proteins over basal level, as determined by GTPase activity (EC50=32+/-8 nM) and by the guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding rate (EC50=61+/-21 nM). The affinity of receptors for progesterone was substantially decreased in the presence of GTPgammaS and of cholera toxin. Our results suggest the existence of progesterone receptors in the membrane of Rhizopus nigricans and their coupling to G proteins.  相似文献   

14.
Previously reported methods for quantifying platelet-activating factor (PAF) binding to rabbit platelet membranes were modified for studies of PAF binding to human platelet membranes. The membranes were prepared by the "glycerol lysis" method and PAF binding was quantified by using polyethylene glycol precipitation to recover membrane-bound PAF. Optimal PAF binding required buffers containing 3 to 10 mm KCl and either 5 to 10 mM MgCl2 or 5 to 10 mM CaCl2. NaCl was not as effective as KCl and concentrations of NaCl greater than 3 mM strongly inhibited PAF binding. Maximal binding occurred after incubation for 60 min at 0 degree C and was reversed by the addition of excess unlabeled PAF. PAF binding was saturable. Scatchard analysis of PAF binding to 50 micrograms of membrane protein revealed 10.3 +/- 1.7 x 10(11) receptors per milligram of membrane protein and the receptors had a Kd of 7.6 +/- 1.9 nM. The calculated receptor number, binding affinity, and specificity of binding are similar to those previously calculated for PAF binding to intact human platelets, suggesting that the membrane binding site for PAF is the PAF receptor.  相似文献   

15.
The protective effect exerted by BN 52021 a specific PAF-receptor antagonist in experimentally induced ocular inflammatory disorders led us to investigate the possible presence of specific receptors for PAF in rabbit iris and ciliary body. Two classes of PAF binding sites were found in isolated iris and ciliary process of pigmented rabbit eyes: a high affinity site Kd1 congruent to 4.9 +/- 0.47 nM, Bmax1 congruent to 3.17 +/- 0.50 pmoles/mg protein, a low affinity sites Kd2 congruent to 11.6 +/- 0.33 nM, Bmax2 congruent to 12.46 +/- 2.3 pmoles/mg protein for iris. The specific binding was not affected by lyso-PAF the biologically inactive precursor and metabolite of PAF, up to 10(-6) M; inhibition by unlabelled PAF demonstrated a biphasic curve partially antagonized by BN 52021. The present results demonstrate the presence of specific binding sites for PAF in rabbit eyes which could mediate the action of this mediator in eye inflammatory processes and explain the protective effect observed with BN 52021.  相似文献   

16.
We have used three experimental protocols to determine binding parameters for type I and type II glucocorticoid receptors in the spinal cord and hippocampus (HIPPO) from adrenalectomized rats. In protocol A, 0.5-20 nM [3H]dexamethasone (DEX) was incubated plus or minus a 1000-fold excess of unlabeled DEX, assuming binding to a two-site model. In protocol B, [3H]DEX competed with a single concentration of RU 28362 (500 nM), whereas in protocol C, we used a concentration of RU 28362 which varied in parallel to that of [3H]DEX, such as 500 x. Results of protocols A and C were qualitatively similar, in that: (1) Bmax for type I receptors favored the HIPPO, while the content of type II sites was comparable in the two tissues; (2) Kd was consistently lower for type I than for type II sites in both tissues; and (3) type II receptors from the spinal cord showed lower affinity than their homologous sites from HIPPO. This last result was also obtained when using protocol B. In contrast, protocol B yielded binding data indicating that type II sites were of similar or higher affinity than type I sites. Computer simulation of the binding protocols demonstrated that protocols A and C were the most theoretically reliable for estimating the Kd and Bmax of type I sites, and the predicted error was smaller for protocol C, in comparison with protocol B. We suggest that the noted differences in the Kd of type II receptors between the spinal cord and HIPPO could account for a difference in sensitivity of the two systems in the physiological adrenal hormone range.  相似文献   

17.
人脊髓创伤后神经元病变的神经丝免疫组织化学研究   总被引:5,自引:0,他引:5  
本文用神经丝(NF)免疫组织化学方法在15例人体尸检材料中研究了脊髓创伤后生存2h~9W的脊髓神经元胞体和轴突的病理学改变。结果表明:脊髓创伤后2h,神经丝免疫组织化学反应即可显示NF阳性反应产物在轴突内聚集。创伤后第4天,病变的前角运动神经元胞体内神经丝反应异常地增强。以上结果表明:神经丝免疫组织化学方法比常规显示轴突的染色方法能更早更清晰地显示脊髓内轴突的病变,并进一步证实了创伤后细胞骨架紊乱在神经元的病理发病机理中起重要作用。  相似文献   

18.
1. Glucocorticoid hormones affect several functions of the spinal cord, such as synaptic transmission, biogenic amine content, lipid metabolism, and the activity of some enzymes (ornithine decarboxylase, glycerolphosphate dehydrogenase), indicating that this tissue is a target of adrenal hormones. 2. Corticosterone, the main glucocorticoid of the rat, is detected at all regional levels of the spinal cord, and cold stress increases this steroid, predominantly in the cervical regions. 3. Intracellular glucocorticoid receptors have been found in the spinal cord, with higher concentrations in the cervical and lumbar enlargements. Prima facie, these receptors presented biochemical, stereospecifical, and physicochemical properties similar to those of receptors found in other regions of the nervous system. The prevalent form in the spinal cord is the type II receptor, although type I is also present in small amounts. 4. The type II glucocorticoid receptor of the spinal cord shows an affinity lower (Kd 3.5 nM) than that of the hippocampal type II site (Kd 0.7 nM) when incubated with [3H]dexamethasone. This condition may impair the nuclear translocation of the spinal cord receptor. 5. Another peculiar property of spinal cord type II site is a greater affinity for DNA-cellulose binding than the hippocampal receptor during heat-induced transformation. Also, the spinal cord receptor shows resistance to the action of RNAse A, an enzyme which increases DNA-cellulose binding of the hippocampal receptor, indicating that both receptors may be structurally different. 6. Therefore, it is possible that a different subclass of type II, or "classical glucocorticoid receptor," is present in the spinal cord. This possibility makes the cord a useful system for studying diversity of glucocorticoid receptors of the nervous system, especially the relationship between receptor structure and function.  相似文献   

19.
High affinity receptors have been demonstrated for the potent phospholipid autacoid, platelet-activating factor (PAF C18:0; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine) in a variety of tissues, including the endometrium. Because of the relative instability of PAF and our previous demonstration that lyso-PAF (1-O-alkyl-2-lyso-sn-glycero-3-phosphorylcholine), the major metabolite of PAF, displaced [3H]PAF from endometrial PAF receptor sites, we have examined the ability of bovine serum albumin (BSA) to prevent degradation of PAF and have characterized PAF and lyso-PAF binding sites in purified rabbit endometrial membranes isolated on Day 6 of pregnancy. In buffer containing the phospholipase A2 inhibitors, quinacrine (10 microM) and dibromoacetophenone (2 microM), and 0.25% BSA, 87.4 +/- 3.2% of added [3H]PAF C18:0 remained intact after incubation at 25 degrees C for 150 min. The metabolic products, lyso-PAF and 1-O-alkyl-2-acyl-sn-glycero-3-phosphorylcholine (alkylacyl-GPC), only amounted to 5.2 +/- 3.2 and 3.3 +/- 1.1, respectively. At the same concentration, rabbit serum albumin (RSA) also significantly protected [3H]PAF C18:0 from metabolism, but bovine gamma globulin (BGG) was ineffective. The presence of 0.25% BSA, however, did not protect [3H]lyso-PAF C18:0 from extensive catabolism: the major product formed was [3H]alkylacyl-GPC. Insignificant amounts of [3H]PAF were formed. Under the same conditions (25 degrees C, 150 min) in the presence of 0.25% BSA, saturation analysis revealed the presence of two types of PAF C18:0 receptors in the endometrial membranes. Type 1 sites had a Kd of 0.42 +/- 0.03 nM (mean +/- SD; n = 3) and binding capacity of 0.11 +/- 0.01 pmol/mg protein. Type 2 receptor sites had a Kd of 5.96 +/- 0.35 nM and a binding capacity of 1.59 +/- 0.22 pmol/mg protein. Thus, in the presence of BSA, the binding capacities of the two classes of receptors were markedly reduced compared to values generated previously in its absence. The Kd of the Type 1 sites was not significantly changed by the presence of BSA. A single class of saturable high-affinity binding sites was demonstrable for lyso-PAF C18:0: Kds ranged from 0.76 +/- 0.58 to 11.1 +/- 0.62 nM, depending on which method of analysis was used (Eadie-Hofstee, Scatchard-Rosenthal, or the Lundon nonlinear method). The binding capacities were equally varied, ranging from 0.15 +/- 0.08 to 15.17 +/- 4.95 pmol/mg protein.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

20.
探讨丝裂原活化蛋白激酶(Mitogen activated protein kinase,MAPK)相关蛋白丝裂原活化蛋白激酶磷酸酶(Mitogen activated protein kinase phoshatase-1,MKP-1)和磷酸化细胞外信号调节激酶(Extracellular sigIlal-regulated kinases,ERK)在大鼠脊髓损伤后表达的变化及其意义.20只SD大鼠随,机分为实验组及假手术对照组.实验组采用改良Allen'S打击法制作脊髓损伤动文为实验组及假手术对照组同法暴露脊髓,但不损伤脊髓.2组大鼠术后12h取手术段脊髓,用苏木精--伊红染色观察损伤脊髓组织病理变化和检测脊髓标本损伤段的MKP-1和磷酸化ERK蛋白表达的差异.实验组脊髓HE染色显示存在大量出血坏死后形成的囊腔,组织和神经细胞水肿以及神经纤维溶解消失.免疫组化和Western Blot结果发现.术后第12h实验组MKP-1蛋白的表达减少,同时磷酸化ERK-1蛋白的表达量却明显增加,差别有显著性意义(P<0.01).脊髓组织受重物打击后可下调MKP-1蛋白的表达,同时显著增加磷酸化ERK蛋白,而这可能是脊髓损伤的机制之一.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号