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Toosendanin increases free-Ca2. concentration in NG108-15 cells υia L-type Ca2. channels

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Acta Pharmacologica Sinica 2004年第5期 55-59页

作者： tong-hui xu, jun ding,{2., shanghai institutes for biological sciences,chinese Academy of sciences, shanghai 2.0031, china

AIM: To examine if toosendanin (TSN) affects intracellular free-Ca2. concentration ([Ca2.]i) in neuroblastomagliomahybrid cells (NG108-15 cells). METHODS: The [Ca2.]i was determined by laser-scanning confocal microscopicimaging technique in which Fluo-3 was used as Ca2. indicator. RESULTS: TSN induced an increase in resting[Ca2.]i and in high K+-evoked Ca2. transient in differentiated NG108-15 cells. The TSN-induced increase in [Ca2.]iwas dose-dependent and disappeared in CdCl2., nifedipine-containing or Ca2.-free solution, and appeared afterwashing out the Ca2. channel blockers or adding Ca2.. CONCLUSION: TSN increased [Ca2.]i in differentiatedNG108-15 cells. The [Ca2.]i enhancement was due to the influx of extracellular Ca2. and related to L-type Ca2.channels.

关键词： toosendanin intracellular calcium concentration confocal microscopy calcium indicator neuroblastomaglioma hybrid cells L-type Ca2. channels

A patch-clamp study on human sperm Cl^- channel reassembled into giant liposome

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Asian Journal of Andrology 2001年第3期3卷 185-191页

作者： jun-Ping BAI yu-{2. shi Key laboratory of neurobiology,Intitute of Physiology,shanghai institutes for biological sciences,chinese Academy of Science,{12.,china Key laboratory of {2. Institute of Physiology Chinese Academy of Sciences Shanghai.

Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp ***: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 2.0 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2., 2.-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2.01 Sep; 3: 185 - 191)

关键词： Cl^-channel giant liposome patch clamp human sperm membrane ion channel reassembly

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Opioid receptor antagonists increase [Ca2.]i in rat arterial smooth muscle cells in hemorrhagic shock

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Acta Pharmacologica Sinica 2004年第3期25卷 141-146页

作者： Li KAI,Zhong-feng WANG2.yu-liang {2.2.3,liang-ming LIU,De-yao HU Institute of Surgery,Department of Pharmacology,Third Military Medical University,Chongqing 400042.Key laboratory of 2.neurobiology,shanghai institutes for biological sciences,chinese Academy of sciences,shanghai 2.0031,china Institute of Surgery Department of Pharmacology Third Military Medical University 重庆 400042 中科院上海生命科学院 Key Laboratory of Neurobiology CAS 上海 200031

AIM: To examine the effects of opioid receptor antagonists and norepinephrine on intracellular free Ca2. concentra- tion ([Ca2.]i) in mesenteric arterial (MA) smooth muscle cells (SMC) isolated from normal and hemorrhagic shocked rats in the vascular hyporesponse stage. METHODS: The rat model of hemorrhagic shock was made by withdrawing blood to decrease the artery mean blood pressure to 3.73-4.2. kPa and keeping at the level for 3 h. [Ca2.]i of vascular smooth muscle cells (VSMC) were detected by the laser scan confocal microscopy. RESULTS: In the hyporesponse VMSC of rats in hemorrhagic shock, selective δ-, κ-, and μ-opioid receptor antagonists (naltrindole, nor-binaltorphimine, and β-funaltrexamine, 100 nmol/L) as well as norepinephrine 5 μmol/L significantly increased [Ca2.]i by 47 %±13 %, 37 %±14 %, 33 %±10 %, and 54 %±17 %, respectively, although their effects were lower than those in the normal rat cells (the increased values were 148 %±54 %, 130 %±44 %, 63 %±17 % and 110 %±38 %, respectively); and the norepinephrine-induced increase in [Ca2.]i was further augmented by three δ-, κ-, and μ-opioid receptor antagonists (50 nmol/L, respectively) application (from 52.%±16 % to 99 %±2. %, 146 %±54 % and 137 %±47 %, respectively). CONCLUSION: The disorder of [Ca2.]i regulation induced by hemorrhagic shock was mediated by opioid receptor and α-adrenoceptor, which may be partly responsible for the vascular hyporesponse, and the opioid receptor antagonists improved the response of resistance arteries to vascu- lar stimulants in decompensatory stage of hemorrhagic shock.

关键词： hemorrhagic shock vascular smooth muscle opioid receptors calcium

Serine racemase expression in mouse cerebral cortex after permanent focal cerebral ischemia

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Acta Pharmacologica Sinica 2004年第4期25卷 38-43页

作者： Li-zhenWANG, Xing-zu ZHU2.Department of Pharmacology, shanghai Institute of Materia Medica, shanghai institutes for biological sciences, {2. Academy of sciences, shanghai 2.12.3, china Department of Pharmacology Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 上海 201203

AIM: To study the alterations of the expressions of serine racemase in C57BL/6 mouse brain after permanent focal cerebral ischemia. METHODS: The mRNA level and the protein level of serine racemase were assayed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. The amount of D-serine and L-serine were measured by HPLC. RESULTS: High levels of serine racemase were constitutively expressed in the normal cortex of mouse. At early stage after middle cerebral artery occlusion (MCAO), no significant change in expression of serine racemase was observed in temporoparietal cortex in ipsila- teral hemisphere. However, delayed transient decreases of serine racemase in both mRNA and protein levels were detected from d 6 to d 10 after ischemia. Correspondingly, D-serine concentration also declined in the ipsilateral cortex during this period when compared with the D-serine level in the contralateral cortex. CONCLUSION: Delayed decreases in serine racemase expression and D-serine level occurred in the temporoparietal cortex at the late stage after focal cerebral ischemia.

关键词： D-serine L-serine serine racemase inbred C57BL mice brain ischemia

Rational redesign of inhibitors of furin/kexin processing proteases by electrostatic mutations

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Acta Pharmacologica Sinica 2004年第12期25卷 146-152页

作者： Xiao-hui CAI, Qing ZHANG, Da-fu ding2.K{2. laboratory of Proteomics, Institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese Academy of sciences, Graduate School of the chinese Academy of sciences, shanghai 2.0031, china 中科院上海生命科学院 Key Laboratory of Proteomics CAS 上海 200031

AIM: To model the three-dimensional structure and investigatethe interaction mechanism ofthe proprotein convertase furin/kexin and their inhibitors (eglin c mutants). METHODS: The three-dimensional complex structures of furin/ kexin with its inhibitors, eglin c mutants, were generated by modeller program using the newly published X-ray crystallographical structures of mouse furin and yeast kexin as templates. The electrostatic interaction energy of each complex was calculated and the results were compared with the experimentally determined inhibition con- stants to find the correlation between them. RESULTS: High quality models of furin/kexin-eglin c mutants were obtained and used for calculation of the electrostatic interaction energies between the proteases and their inhibitors. The calculated electrostaticenergies of interaction showeda linear correlation to the experimental inhibition constants. CONCLUSION: The modeled structures give good explanations of the specificity of eglin c mutants to furin/ kexin. The electrostatic interactions play important roles in inhibitory activity of eglin c mutants to furin/kexin. The results presented here provided quantitative structural and functional information concerning the role of the charge- charge interactions in the binding of furin/kexin and their inhibitors.

关键词： furin kexin eglin c drug design electrostatic mutation

Inducible nitric oxide synthase contributes to intermittent hypoxia against ischemia/reperfusion injury

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Acta Pharmacologica Sinica 2005年第3期26卷 315-322页

作者： Hai-lei ding~2.Hai-feng ZHU~2.Jian-wen DONG~2.Wei-zhong ZHU~2.Wei-wei YANG~3 Huang-tian YANG~3 Zhao-nian ZHOU~(2.4)2.Physiological laboratory of Hypoxia,{2. institutes for biological sciences,chinese Academy of sciences,{2. 2.0031,china 3 laboratory of Molecular Cardiology,Health Science Center,{2. institutes for biological sciences,chinese Academy of sciences and {2. Second Medical University,{2. 2.002.,china Physiological laboratory of Hypoxia Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China laboratory of Molecular Cardiology Health Science Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Shanghai Second Medical University Shanghai China

Aim:To investigate the role of inducible nitric oxide synthase (iNOS)-derivednitric oxide (NO) in the cardioprotection of intermittent hypoxia (IH) against is-chemia/reperfusion (I/R) ***:Langendorff-perfused isolated rathearts were used to measure variables of left ventricular function during baselineperfusion,ischemia,and reperfusion *** plus nitrite (NOx) content inmyocardium was measured using a biochemical method,iNOS mRNA and pro-tein expression in rat left ventricles were detected using reverse transcription poly-merase chain reaction (RT-PCR) and Western blot,***:Myo-cardial function recovered better in IH rat hearts than in normoxic control *** iNOS-selective inhibitor aminoguanidine (AG) (100μmol/L) significantlyinhibited the protective effects of IH,but had no influence on normoxic rat *** baseline content of NOx in IH hearts was higher than that in normoxic *** 30 min ischemia,the NOx level in normoxic hearts increased compared tothe corresponding baseline level,whereas there was no significant change in ***,the NOx level in IH hearts was still higher than that of normoxichearts during ischemia and reperfusion *** 100μmol/L significantly di-minished the NOx content in IH and normoxic hearts during ischemia and *** baseline levels of iNOS mRNA and protein in IH hearts were higherthan those of normoxic *** to the corresponding baseline level,iNOS mRNA and protein levels in normoxic rat hearts increased and those in IH rathearts decreased after *** addition of AG 100μmol/L significantlydecreased iNOS mRNA and protein expression in IH rat hearts after I/***:IH upregulated the baseline level of iNOS mRNA and protein expres-sion leading to an increase in NO production,which may play an important role inthe cardiac protection of IH against I/R injury.

关键词： intermittent hypoxia ischemia/reperfusion inducible nitric oxide synthase aminoguanidine: rats

Toosendanin interferes with pore formation of botulinum toxin type A in PC12.cell membrane

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Acta Pharmacologica Sinica 2006年第1期27卷 66-70页

作者： Mu-feng LI yu-liang {2. Institute of Physiology,shanghai institutes for biological sciences,chinese Academy of sciences,shanghai 2.0031,china Institute of Physiology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim:Botulinum neurotoxins (BoNT) abort the process of neurotransmitter re-lease at presynaptic motor nerve terminals,causing muscle *** abilityof botulinum toxin to produce its effect is dependent on the ability of the lightchain to cleave the SNARE proteins associated with transmitter ***-cation of the light chain protease through the heavy chain-formed channel is apivotal step in the intoxication *** (TSN),a triterpenoid de-rivative extracted from a chinese traditional medicine,has been demonstrated tobe an effective cure for experimental *** study was designed to explorethe antibotulismic mechanisms of ***:The inside-out single-channel recording patch-clamp technique was used to record the BoNT/A-in-duced currents in the presence and absence of ***:Channel formationwas delayed and the sizes of the channels were reduced in the TSN-treated PC12.ell ***:The antibotulismic effect of TSN might occur viainterference with toxin translocation.

关键词： toosendanin botulinum toxin type A ion channel antibotulismic mechanism single channel recording

The Role of Propeptide in the Refolding of Human Group IB Phospholipase A2

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 583-588页

作者： Hong-Qiang CHENG1 and Gen-jun xu1,2.Institute of Biochemistry and Cell Biology,{2., chinese Academy of sciences, shanghai 2.0031, china 2.ollege of Life Science, Zhejiang University of sciences, Hangzhou 310033, china Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China College of Life Science Zhejiang University of SciencesHangzhou 310033 China

Human group IB phospholipase A2.(IB-PLA2. and its zymogen (proIB-PLA2. were purified from E. coli. Refolding was carried out by diluting the denatured forms of both IB-PLA2.and proIB-PLA2. with renaturation buffer in which the disulfide bonds were completely reduced. The refolding yield of proIB-PLA2.was increased by about 50% over that of the mature enzyme. The refolding of IB-PLA2.usually produced aggregates under normal conditions, as determined by light scattering. In addition, the unfolding experiments showed that the mature enzyme was more stable than the proenzyme toward denaturants in the presence of DTT. Results suggested that the N-terminal sequence rather than its conformation of human proIB-PLA2.played an important role in the refolding process.

关键词： human group IB phospholipase A2.(IB-PLA2. refolding first disulfide bond propeptide

Computational neuroscience in china

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Science china(Life sciences) 2010年第3期53卷 385-397页

作者： WU Si1 & liang PeiJi2.1Institute of Neuroscience,shanghai {2. for biological sciences,State Key laboratory of neurobiology,chinese Academy of sciences,shanghai 2.0031,{12. 2.epartment of Biomedical Engineering,shanghai Jiaotong University,shanghai 2.02.0,{12. Institute of Neuroscience Shanghai Institutes for Biological Sciences State Key Laboratory of Neurobiology Chinese Academy of Sciences Shanghai China Department of Biomedical Engineering Shanghai Jiaotong University Shanghai China

The ultimate goal of Computational Neuroscience(CNS) is to use and develop mathematical models and approaches to elucidate brain *** is a young and highly multidisciplinary *** heavily interacts with experimental neuroscience and such other research areas as artificial intelligence,robotics,computer vision,information science and machine *** paper reviews the history of CNS in china,its current status and the prospects for its future *** of CNS research in china are also presented.

关键词： computational neuroscience experimental neuroscience artificial intelligence

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