钙及钙调素研究进展(Ⅰ)

解决细胞识别、细胞通讯、细胞代谢的调控、细胞增殖与细胞周期、细胞与抗病以及细胞对外界信号进行的持续反应维持等重大理论与实际问题。文中着重介绍了作为胞内信使的钙离子及其循环的重要意义、作为Ca~(2+)受体的胞内蛋白CaM的广泛性。作者认为CaM在细胞代谢调控中处于中心位置,对Ca~(2+)及其循环、第二信使系统CAMP和肌醇磷脂,甚致CGMP均有调节、协调作用;是调控细胞代谢的综合剂。     

Protein kinase C regulation of GABAA receptors

Pharmacological studies with drugs that activate or inhibit several protein kinase C (PKC) isozymes have identified the PKC family of serine-threonine kinases as important in the regulation of -aminobutyric acid type A (GABA_A) receptor function. PKC modulates GABA_A receptor surface density, chloride conductance and receptor sensitivity to positive allosteric modulators such as neurosteroids, ethanol, benzodiazepines and barbiturates. Recent studies using PKC isozyme-selective reagents and gene-targeted mice have begun to identify critical roles for three isozymes, PKCII, PKC and PKC, in various aspects of GABA_A receptor regulation. Progress in this field touches upon therapeutic areas that are of great clinical importance such as anxiety and addiction. Increased understanding of how PKC regulates GABA_A receptors and which PKC isozymes are involved holds promise for development of new treatments for diverse neuropsychiatric disorders.Received 2 August 2004; received after revision 17 August 2004; accepted 21 August 2004     

RACK1依赖的miRNA途径参与调节植物叶绿素的生物合成

为深入探索活化C激酶受体1(receptor of activated C kinase 1,RACK 1)在调节植物microRNA(miRNA)的生物发生,以及其靶基因中的关键作用,对在美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)网站上共享的基因表达综合数据库(Gene Expression Omnibus,GEO)中有关rack1突变体的小RNA序列数据重新进行了系统分析和挖掘,找到了19个新miRNA.通过解析差异miRNA表达,鉴定到了特异调控叶绿素合成相关基因HEMA和HEMC表达的miRNA,为今后深入系统地研究RACK1在调节叶绿体发育和光合作用机理提供了关键的理论依据,也为利用公共数据库挖掘潜在的数据信息提供了基本的研究设想和思路.     

牛磺酸抗心肌成纤维细胞增殖作用的实验研究

目的研究牛磺酸(Taurine,Tau)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导新生大乳鼠心肌成纤维细胞(myofibroblasts,myo Fbs)增殖的抑制作用,并探讨其作用机制.方法用AngⅡ诱导新生大乳鼠myo Fbs增殖,建立心肌纤维化(myofibrosis,MF)模型.采用四甲基偶氮唑盐(MTT)法检测细胞增殖;羟脯氨酸试剂盒检测胶原含量;免疫细胞化学染色检测p-PKCα的膜转位及表达;Western blot检测p-PKCα和p-ERK1/2蛋白表达及含量.结果 Tau(30,60,120)mmol/L可明显抑制AngⅡ诱导的Myo Fbs增殖及胶原合成,同AngⅡ组比较差异具有统计学意义(P0.05或P0.01),并且呈现出剂量依赖性.应用免疫细胞染色Western blot并且结合图像分析,结果表明:Tau可抑制p-PKCα膜转位和表达,与AngⅡ组相比差异具有统计学意义(P0.01).结论 Tau可能通过减少p-PKCα的表达及膜转位,从而抑制MyoFbs增殖和胶原含量的增加,抑制MF,逆转心肌重塑.     

Expression, distribution and function of the focal adhesion kinase (pp125FAK) during murine ectoplacental cone outgrowthin vitro

Mouse embryo implantation is a complex process that includes trophoblast cells derived from ectoplacental cone (EPC) adhesion to and migration through the extracellular matrix (ECM) of uterine endometrium and invasion into the decidua. At the time of implantation, fibronectin (FN) is abundant in the decidua and is distributed pericellularly around each individual stromal cell, and its receptor (integrin α-5β-1) expression on trophoblast populations is up-regulated. The focal adhesion kinase, a 125 ku protein tyrosine kinase (pp125 FAK), is tyrosine phosphorylated upon integrin engagement with its ECM ligand, and its tyrosine phosphorylation sites then serve as the binding sites which couple it with cellular proteins that contain Src SH2 or SH3 domains. Through these linkages, pp125 FAK may integrate multiple signals triggered by integrins. The model of EPC culture %in vitro% was used to study the expression, distribution and function of pp125 FAK during EPC outgrowth on FN. Results indicated that, pp125 FAK primarily expressed and distributed in cellular focal adhesions of the front edge of trophoblast outgrowth from EPC, and was localized in the peripheral region of the individual migrating trophblast cell; antibody or antisense oligodeoxynucleotide to pp125 FAK inhibited EPC attachment and outgrowth, as well as trophoblast cells spreading and migration. This experiment demonstrated that pp125 FAK as an integrin-mediated signaling molecule was involved in EPC outgrowth %in vitro%, and played an important role during trophoblast cells interaction with FN.     

酵母(Saccharomyces cerevisiae)Sch9蛋白激酶PDK1位点体内磷酸化的研究

根据Sch9氨基酸序列中激活区570位苏氨酸（T570）位点，也被称为PDK1位点附近的氨基酸序列设计了一段磷酸化多肽，并获得了570位磷酸化苏氨酸特异性抗体. 实验表明该抗体可有效区别Sch9 PDK1位点的磷酸化和非磷酸化. 使用该抗体检测生理条件下表达的Sch9蛋白，发现Sch9的PDK1位点在生理条件下发生了明显的磷酸化.