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41.
42.
SH2A 基因对细胞信号转导的影响及其亚细胞定位 总被引:3,自引:0,他引:3
目的 研究Src同源域2(src homology 2,SH2)A基因在细胞信号转导中的作用并进行亚细胞定位。方法 通过RT—PCR方法扩增SH2A cDNA编码序列,构建真核重组表达载体pcDNA3.1-SH2A,利用脂质体转染肝癌Bel7402细胞、COS7细胞,检测蛋白酶C(protein kinaseC,PKC)、酪氨酸蛋白激酶(tyrosine protein kinase,TPK)、丝裂原激活蛋白激酶(mitogen activated protein kinase,MAPK)活性的改变;另构建pEGEP—SH2A,转染同前,荧光显微镜观察荧光定位。结果 测序结果显示真核重组表达载体pcDNA3.1-SH2A及pEGFP—SH2A中均含有SH2AcDNA编码序列;肝癌Bel7402细胞、COS7细胞转染pcDNA3.1-SH2A后,胞浆PKC的活性下降了40%左右,MAPK和TPK活性未见明显改变。荧光显微镜观察发现SH2A基因在细胞质中表达。结论 SH2A基因编码蛋白在PKC信号转导通路中起抑制作用;SH2A基因编码蛋白定位于细胞质。 相似文献
43.
A Parametric Approach to Measuring Cerebral Blood Flow Autoregulation from Spontaneous Variations in Blood Pressure 总被引:2,自引:0,他引:2
Autoregulation maintains cerebral blood flow (CBF) almost constant in the face of changes in arterial blood pressure (ABP). Tests for impairment of this process using only spontaneous fluctuations in ABP, without provoking large variations, are of great clinical interest, and a range of different approaches have previously been applied. Extending earlier work based on linear filters, we propose a simple parametric method using a first order finite impulse response filter. We evaluate the method on ABP and CBF velocity [(CBFV), from trancranial Doppler ultrasound] signals collected in 60 patients with stenosis or occlusion of the carotid arteries. Data were collected during the inspiration of ambient air, a 5% CO2/air mixture, and finally the return to ambient air. Equivalent data were collected in 15 normal subjects. The filters estimated from the data segments with constant inspiratory pCO2 showed the expected high-pass characteristic, which was reduced during hypercapnia and also in patients. Highly significant correlation between the filter parameters and cerebrovascular reactivity (percent increase in CBFV per unit change in end-tidal pCO2) gives further evidence that the filters reflect autoregulation. The method allows simple parametrization of the dynamic autoregulatory responses in CBFV, and the analysis of short (1 min) data segments. © 2001 Biomedical Engineering Society.
PAC01: 8719Uv, 4762+q, 4380Qf 相似文献
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45.
上皮间质转化(EMT)及其分子机制 总被引:7,自引:0,他引:7
上皮细胞受到细胞外因子刺激后向间质细胞转化的现象与肿瘤的浸润转移密切相关。在此过程中上皮细胞的极性丧失,迁移和运动能力增强,同时上皮表型丢失而逐渐获得间质表型。参与这一过程的细胞内信号转导途径有受体酪氨酸激酶Ras-MAPK途径,Src激酶,Rho家族激酶,PI3K/AKT途径,Wnt信号通路,转录因子等。 相似文献
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47.
The productive activation of CD4(+) T lymphocytes, leading to proliferation and cytokine secretion, requires precise temporal regulation of intracellular cyclic AMP concentrations. The major effector molecule activated by cyclic AMP in mammalian cells is the cyclic AMP-dependent protein kinase A (PKA). The type I PKA isozyme mediates the inhibitory effects of cyclic AMP on T-cell activation. Using laser scanning confocal microscopy, we demonstrated that the regulation of PKA type I activity involves spatial redistribution of PKA type I molecules following T-cell receptor (TCR) stimulation. In resting T cells, PKA type I was located in membrane proximal regions and distributed equally across the cell. Shortly after antigen engagement, T cells and antigen-presenting cells formed an area of intense contact, known as the immunological synapse. TCR concentrated at the synapse, whereas PKA type I molecules redistributed to the opposite cell pole within 10 min after T-cell stimulation. Type I PKA redistribution was solely dependent on TCR signalling, because we observed the same temporal and spatial distribution after antibody-mediated cross-linking of the TCR-associated CD3 complex. Segregation of TCR and PKA type I molecules was maintained for at least 20 min. Thirty minutes after stimulation, PKA type I partially colocalized with the TCR. After 60 min, PKA type I distribution again approached the resting state. Considering that initial TCR signals lead to increases in intracellular cyclic AMP, PKA type I molecules may be targeted towards localized cyclic AMP accumulations or transported away from these areas, depending on the requirements of the cellular response. 相似文献
48.
Putney JW 《Pflügers Archiv : European journal of physiology》2005,451(1):29-34
TRPC (canonical transient receptor potential) channels are vertebrate homologs of the Drosophila photoreceptor channel, TRP. Considerable research has been brought to bear on the seven members of this family, especially with regard to their possible role in calcium entry. Unfortunately, the current literature presents a confusing picture, with different laboratories producing widely differing results and interpretations. It appears that ectopically expressed TRPC channels can be activated by phospholipase C products (generally, diacylglycerols), by stimulation of trafficking to the plasma membrane, or by depletion of intracellular Ca2+ stores. Here, I discuss the possibility that these diverse experimental findings arise because TRPC channels can, under both experimental as well as physiological conditions, be activated in three distinct ways, possibly depending on their subunit composition and/or signaling complex environment. The TRPCs may be unique among ion-channel subunit families in being able to participate in the assembly and function of multiple types of physiologically important ion channels. 相似文献
49.
目的:在钙化大鼠主动脉血管平滑肌细胞上观察血管紧张素-(1-7)[Angiotensin-(1-7)]对钙化的影响及其信号通道。方法:用β-磷酸甘油制备钙化的大鼠血管平滑肌细胞,再以血管紧张素-(1-7)、血管紧张素Ⅱ、血管紧张素Ⅱ 血管紧张素-(1-7)、选择性蛋白激酶A(PKA)或蛋白激酶C(PKC)抑制剂等干预,通过Von Kossa染色及检测钙含量、碱性磷酸酶活性、骨钙素浓度和Cbfa1 mRNA表达来探讨血管紧张素Ⅱ对钙化的影响及其信号通道。结果:血管紧张素-(1-7)抑制钙化大鼠血管平滑肌细胞的钙含量、碱性磷酸酶活性(P>0.05)、骨钙素浓度和Cbfa1 mRNA表达(P<0.05),也抑制血管紧张素Ⅱ对血管平滑肌细胞的钙含量、碱性磷酸酶活性、骨钙素浓度和Cbfa1 mRNA表达的促进作用(P<0.05);血管紧张素-(1-7)提高血管平滑肌细胞内cAMP浓度(P<0.05),PKA抑制剂可阻断血管紧张素-(1-7)对钙化血管平滑肌细胞的钙含量、碱性磷酸酶活性、骨钙素浓度和Cbfa1 mRNA表达的抑制作用(P<0.05)。结论:血管紧张素-(1-7)可抑制β-磷酸甘油诱导的血管平滑肌细胞钙化,并拮抗血管紧张素Ⅱ促进的血管平滑肌细胞钙化;这些效应与cAMP-PKA-Cbfa1信号途径有关。 相似文献
50.
Straube E Knöfel B Schneider T Schubert S Wetzker R 《International journal of medical microbiology : IJMM》2000,290(1):65-73
Adhesion to and internalization into host cells is an essential step in the pathogenesis of various bacterial infections. Here we investigated the effects of growth factors on the internalization of Escherichia coli O18 strains isolated from patients with urinary tract infection (UTI) by human epithelial cells. A dramatic increase in the uptake of Escherichia coli was observed after treatment of epithelial cells with epidermal growth factor (EGF) and to a lower extent with insulin. EGF-dependent internalization can be suppressed by tyrosine kinase inhibitors suggesting an involvement of the receptor tyrosine kinases in the regulation of the endocytotic process. Inhibitors of phospholipase A2, lipoxygenase, and cyclooxygenase significantly decreased internalization of bacteria induced by EGF. Finally, the specific inhibitor of PI 3-kinases Wortmannin was shown to suppress completely the EGF-independent internalization. The data of this analysis indicate the involvement of several signaling paths in bacterial internalization of uropathogenic Escherichia coli O18 strains and contribute to the comprehension of the pathogenesis of recurrent UTI. 相似文献