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ONO-1078 reduces NMDA-induced brain injury and vascular cell adhesion molecule-1 expression in rats

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Acta Pharmacologica Sinica 2005年第4期26卷 435-440页

作者： Li-hui ZHANG~(2,3) Er-qing WEI~(2,4) 2 department of pharmacology,school of Medicine,zhejiang university,hangzhou 310031 3 medical school,hangzhou Normal university,hangzhou 310018,china department of pharmacology School of Medicine Zhejiang University Hangzhou China medical school Hangzhou Normal University Hangzhou China

Aim:To determine whether ONO-1078 （pranlukast）,a potent cysteinyl leukotrienereceptor 1 （CysLTI） antagonist,has an effect on N-methyl-D-aspartate （NMDA）-induced brain injury and vascular cell adhesion molecule-1 （VCAM-1） expres-sion in ***:Brain injury was induced by direct microinjection of NMDA（0.3 μmol in 1 μL of sterile 0.1 mol/L PBS,pH 7.4） into the cerebral *** volume （area）,brain edema and neuron density were assessed by an imageanalyzer and the expression of VCAM-1 in the cortex was detected by Westernblot 24 h after NMDA ***-1078 （0.03,0.1,or 0.3 mg/kg） andedaravone （MCI-186,10 mg/kg）,a neuroprotective agent,were ip injected 30rain before and after NMDA ***:NMDA microinjection producedwell-defined focal lesions （Figure 1） dose- and ***-1078（0.1,0.3 mg/kg） and edaravone （10 mg/kg） decreased the total lesion volume,lesion area and brain edema induced by ***,ONO-1078 （0.1,0.3mg/kg） significantly inhibited the enhanced expression of VCAM-1 in the injuredcortices,but edaravone did not have this ***:CysLT1receptorantagonist ONO-1078 at tenufftes NMDA-induced brain damage in rats,and thismight relate to the attenuation of NMDA receptor-dependent neurotoxicity andthe inhibition of the upregulation of VCAM-1 expression.

关键词： leukotriene antagonists pranlukast N-methylaspartate vascular cell adhesion molecule-1 neurotoxicity brain injuries

Endogenous histamine inhibits the development of morphine-induced conditioned place preference

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Acta Pharmacologica Sinica 2007年第1期28卷 10-18页

作者： Ying-xia GONG~2 Min LV~(2,3) Yong-ping ZHU~2 Yuan-yuan ZHU~2 Er-qing WEI~2 Hong SHI~3 Qun-li ZENG~2 Zhong CHEN~(2,4)2 department of pharmacology and Neurobiology,school of Medicine,zhejiang university,hangzhou 310031,china 3 department of pharmacology,zhejiang Academy of medical Sciences,hangzhou 310031,china department of pharmacology and Neurobiology School of Medicine Zhejiang University Hangzhou China department of pharmacology Zhejiang Academy of Medical Sciences Hangzhou China

Aim:The conditioned place preference(CPP)paradigm was used to investigatethe effects of endogenous histamine on the processes leading to morphine-induced reward-seeking behavior in Sprague-Dawley ***:The modelof CPP was used to assess the rewarding effect of *** levels of histamine,glutamate,γ-aminobutyric acid(GABA),dopamine(DA)and 3,4-dihydroxyphenyl-acetic acid(DOPAC)in rat brains were measured with high-performance *** technique was used to observe themorphological changes of ***:Intraperitoneal injection of mor-phine(2,5 or 10 mg/kg)induced the development of CPP in a *** addition,morphine administrations(10 mg/kg)decreased the hista-mine content and reduced the number and size of histaminergic neurons in thetuberomammillary nucleus(TM),as well as markedly increasing the DOPAC/DAratios in the ventral tegmental area(VTA)and nucleus accumbens(NAc).Intra-peritoneal injection of histidine(50,100 or 200 mg/kg)dose-dependently inhibitedthe development of morphine-induced *** lesions of the TM,whichdecreased the histamine levels in the VTA and NAc,potentiated the developmentof CPP induced by morphine(1 mg/kg,a dose that produced no appreciable effectwhen given alone)and increased the DOPAC/DA ratios in the VTA and NAc,butdid not change the glutamate or GABA levels in these *** reversedthe effects of the TM ***:These results indicate that endog-enous histamine plays a role in inhibiting the development of morphine-inducedreward-seeking behavior,and the inhibition may involve the modulation of dopa-minergic activity.

关键词： conditioned place preference histamine morphine rat tuberomammillary nucleus

Distribution of cysteinyl leukotriene receptor 2 in human traumatic brain injury and brain tumors

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Acta Pharmacologica Sinica 2005年第6期26卷 685-690页

作者： Hua HU~(2,3), Gao CHEN~2, Jian-min ZHANG~2, Wei-ping ZHANG~3, Lei ZHANG~3, Qiu-fu GE~3, Hong-tian YAO~4, WeiDING~4, Zhong CHEN~3, Er-qing WEI~(3,5) ~2 department of Neurosurgery, the Second Affiliated Hospital, school of Medicine, zhejiang university, hangzhou 310009 ~3 department of pharmacology school of Medicine, zhejiang university, hangzhou 310031 ~4 department of Pathology, the First Affiliated Hospital, school of Medicine, zhejiang university, hangzhou 310003, china department of Neurosurgery the Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China department of pharmacology School of Medicine Zhejiang University Hangzhou China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim: To determine the distribution of cysteinyl leukotriene receptor 2 （CysLT2）, one of the cysteinyl leukotriene receptors, in human brains with traumatic injury and tumors. Methods: Brain specimens were obtained from patients who underwent brain surgery. CysLT2in brain tissues was examined using immunohis-tochemical analysis. Results: CysLT2was expressed in the smooth muscle cells （not in the endothelial cells） of arteries and veins. CysLT2was also expressed in the granulocytes in both vessels and in the brain parenchyma. In addition, CysLT2was detected in neuron- and glial-appearing cells in either the late stages of traumatic injury or in the area surrounding the tumors. Microvessels regenerated 8 d after trauma and CysLT2expression was recorded in their endothelial cells. Conclusion: CysLT2is distributed in vascular smooth muscle cells and granulocytes, and brain trauma and tumor can induce its expression in vascular endothelial cells and in a number of other cells.

关键词： cysteinyl leukotriene receptor 2 brain injuries brain neoplasms vascular smooth muscle cell

Cloning of cytochrome P-450 2C9 cDNA from human liver and its expression in CHL cells

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World Journal of Gastroenterology 2002年第2期8卷 318-322页

作者： Ge-Jian Zhu Ying-Nian Yu,department of Pathophysiology and Laboratory of medical Molecular Biology,zhejiang university school of Medicine,hangzhou 310031,zhejiang Province,china Xin Li,department of pharmaceutical analysis & drug metabolism,College of pharmacology Science,zhejiang university,hangzhou 310031,zhejiang Province,china Yu-Li Qian, Present address:Center of laboratory,Women’s hospital,school of Medicine,zhejiang university,hangzhou 310031,zhejiang Province,china department of Pathophysiology and Laboratory of medical Molecular Biology Zhejiang University School of Medicine Hangzhou 310031 Zhejiang Province China.

AIM: Using bacterial, yeast, or mammalian cell expressing a human drug metabolism enzyme would seem good way to study drug metabolism-related problems. Human cytochrome P-450 2C9(CYP2C9) is a polymorphic enzyme responsible for the metabolism of a large number of clinically important drugs. It ranks among the most important drug metabolizing enzymes in humans. In order to provide a sufficient amount of the enzyme for drug metabolic research, the CYP2C9 cDNA was cloned and expressed stably in CHL cells. METHODS: After extraction of total RNA from human liver tissue, the human CYP2C9 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR), and cloned into cloning vector pGEM-T. The cDNA fragment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A transgenic cell line was established by transfecting the recombinant vector of pREP9-CYP2C9 into CHL cells. The enzyme activity of CYP2C9 catalyzing oxidation of tolbutamide to hydroxy tolbutamide in S9 fraction of the cell was determined by high performance liquid chromatography(HPLC). RESULTS: The amino acid sequence predicted from the cDNA segment was identical to that of CYP2C9*1, the wild type CYP2C9. However, there were two base differences, i.e. 21T>C, 1146C>T, but the encoding amino acid sequence was the same, L7, P382. The S9 fraction of the established cell line metabolizes tolbutamide to hydroxy tolbutamide; tolbutamide hydroxylase activity was found to be 0.465 +/- 0.109 ***(-1).g(-1) S9 protein or 8.62 +/- ***(-1).mol(-1) CYP, but was undetectable in parental CHL cell. CONCLUSION: The cDNA of human CYP2C9 was successfully cloned and a cell line of CHL- CYP2C9, efficiently expressing the protein of CYP2C9, was established.

关键词： Cloning, Molecular Animals Aryl Hydrocarbon Hydroxylases Cell Fractionation Cell Line china Gene Expression Humans Hypoglycemic Agents Liver Protein Isoforms Recombinant Proteins Research Support, Non-U.S. Gov't Tolbutamide

Inducible effects of icariin,icaritin,and desmethylicaritin on directional differentiation of embryonic stem cells into cardiomyocytes in vitro

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Acta Pharmacologica Sinica 2005年第4期26卷 477-485页

作者： Dan-yah ZHU Yi-jia LOU~2 department of pharmacology and Toxicology,College of Pharmaceutical Scienees,zhejiang university,hangzhou 310031,china department of pharmacology and Toxicology College of Pharmaceutical Sciences Zhejiang University Hangzhou China

Aim:To investigate the possible inducible effects of icariin,icaritin,anddesmethylicaritin on the directional differentiation of embryonic stem(ES)cellsinto cardiomyocytes in ***:ES cells were cultivated as embryoidbodies(EBs)in hanging drops with icariin,icaritin,or *** cellstreated with retinoic acid and with solvent were used as positive and negativecontrols,*** cardiomyocytes derived from the ES cells were veri-fied using *** expression of cardiac developmental-dependent genes was detected using the reverse transcription-polymerase chainreaction(RT-PCR)*** cycle distribution and apoptosis were analyzedusing flow cytometry to determine the partly inducible effect mechanisms ***:The total percentage of beating EBs treated with 1×10-7mol/L icariin,icaritin,or desmethylicaritin was 87%(P<0.01),59%(P<0.01),and 49%,*** the beating cardiomyocytes derived from the ES cells expressed cardiac-specific proteins for α-actinin and troponin *** them,1×10-7mol/L icariintreatment resulted in a significantly advanced and increased mRNA level ofα-cardiac myosin heavy chain(MHC)and myosin light chain 2v(MLC-2v)in EBsin the early cardiac developmental *** shifting to the cardiomyocytephenotype,icariin could evoke the accumulation of ES cells in G0/G1and acceler-ate apoptosis of the cell population(P<0.05).Conclusion:Icariin facilitated thedirectional differentiation of ES cells into cardiomyocytes at a concentration of1×10-7mol/*** promoting effect of icariin on cardiac differentiation was relatedto increasing and accelerating gene expression of a-cardiac MHC and MLC-2v,aswell as regulating the cell cycles and inducing apoptosis.

关键词： embryonic stein cells directional differentiation cardiomyocytes icariin icaritin desmethylicaritin myosin heavy chain myosin light chain 2 cell cycle apoptosis

Identification of differentially expressed genes in normal mucosa,adenoma and adenocarcinoma of colon by SSH

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World Journal of Gastroenterology 2001年第5期7卷 726-731页

作者： Min-Jie Luo Mao-De Lai department of Pathology,school of Medicine,zhejiang university,hangzhou 310031,P.R.china department of Pathology School of Medicine Zhejiang University Hangzhou 310031 China.

AIM: To construct subtracted cDNA libraries and further identify differentially expressed genes that are related to the development of colorectal carcinoma(CRC). METHODS: Suppression subtractive hybridization(SSH) was done on cDNAs of normal mucosa, adenoma and adenocarcinoma tissues from the same patient. Three subtracted cDNA libraries were constructed and then hybridized with forward and backward subtracted probes for differential screening. Positive clones from each subtracted cDNA library were selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. RESULTS: By this way, there were about 3-4 X 10(2) clones identified in each subtracted cDNA library, in which about 85% positive clones were differentially screened. Sequencing and BLAST homology search revealed some clones containing sequences of known gene fragments and several possibly novel genes showing few or no sequence homologies with any known sequences in the database. CONCLUSION: All results confirmed the effectiveness and sensitivity of SSH. The differentially expressed genes during the development of CRC can be used to shed light on the pathogenesis of CRC and be useful genetic markers for early diagnosis and therapy.

关键词： Gene Expression Regulation, Neoplastic Adenocarcinoma Adenoma Blotting, Northern Colorectal Neoplasms Gene Library Genetic Markers Humans Intestinal Mucosa Polymerase Chain Reaction RNA, Messenger Research Support, Non-U.S. Gov't

Icariin-mediated expression of cardiac genes and modulation of nitric oxide signaling pathway during differentiation of mouse embryonic stem cells into cardiomyocytes in vitro

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Acta Pharmacologica Sinica 2006年第3期27卷 311-320页

作者： Dan-yan ZHU Yi-jia LOU~2 department of pharmacology and Toxicology.College of Pharmaceutical Sciences.zhejiang university,hangzhou 310031.china department of pharmacology and Toxicology College of Pharmaceutical Sciences Zhejiang University Hangzhou China

Aim:To investigate effects of icariin on cardiac gene expression and the modula-tion of nitric oxide(NO)signal transduction during the differentiation of embry-onic stem(ES)cells into cardiomyocytes in ***:The expression levelsof cardiac developmental-dependent genes were measured using reverse tran-scription-polymerase chain reaction(RT-PCR).The chronotropic responses ofcardiomyocytes to β-adrenoceptor stimulation were *** levels ofcAMP and cGMP in ES cells were measured using ***-enous NO levels were measured by using the Griess ***(AG)was used to confirm the influence of icariin on the endogenous NO ***:Icariin significantly elevated mRNA levels of cardiac tran-scription factors GATA4 and Nkx2.5,and cardiac-specific α-MHC,MLC-2v and β-AR genes in a concentration-and time-dependent manner(P35% on d 5+11,an effect that could berescued by *** cells were cocultured with icariin and AG,the percentageof beating EB reached a peak level of 73% on d 5+11(P<0.05),Conclusion:Theinducible effects of icariin were partly related to increase in the expression ofcardiac developmental-dependent genes,and elevation of the cAMP/cGMP ratioin ES cells,as well as upregulation of endogenous NO generation during the earlystages of cardiac development.

关键词： embryonic stem cells cardiomyocytes icariin cardiac-specific genes cAMP cGMP nitric oxide

Inhibitory effect of leflunomide on hepatic fibrosis induced by CCl4 in rats

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Acta Pharmacologica Sinica 2004年第7期25卷 69-74页

作者： Hong-wei YAO2,3, Jun LI3,4, Ji-qiang CHEN2, Shu-yun XU3 2 zhejiang Respiratory Drugs Research Laboratory of State Drugs Administration of china, school of Medicine, zhejiang university, hangzhou 310031 3Institute of Clinical pharmacology, Anhui medical university, Hefei 230032, china 浙江大学国家食品药品监督管理局呼吸药物研究实验室 Hangzhou 310031 Institute of Clinical pharmacology Anhui Medical University Hefei

AIM: To study the effect of leflunomide on CCl4-induced hepatic fibrosis in rats. METHODS: Hepatic fibrosis was induced by subcutaneous injection with 50 % CCl4 in Sprague-Dawley rats. The amount of CCl4 administered was 1 mg/kg. The alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) levels in plasma and hydroxyproline (Hyp) contents in liver tissue were assayed by spectrophotometry. The hyaluronic acid (HA) and procollagen III (PC III) were assessed by radioimmunoassay. The transforming growth factor-β1 (TGF-β1) in serum was determined by ELISA. The nuclear factor-kappa B (NF-κB) in liver tissue was examined by immunohistochemistry. Liver samples collected after 12 weeks of CCl4 treatment were stained with hematoxy- lin and eosin. RESULTS: Leflunomide (1, 3, and 9 mg/kg) significantly decreased indices of liver and spleen, the serum transaminase (AST, ALT) activities, HA and PC III levels, and Hyp contents in liver tissue in rats of hepatic fibrosis. Histopathological examination showed leflunomide had inhibitory effect on fibrogenesis and formation of pseudolobulus. Furthermore, leflunomide significantly inhibited NF-κB expression in liver tissue, and reduced elevated serum TGF-β1 and NO levels in rats of hepatic fibrosis. CONCLUSION: Leflunomide showed inhibitory action on hepatic fibrosis induced by CCl4 in rats.

关键词： leflunomide hepatic fibrosis nuclear factor-kappa B transforming growth factor-beta nitric oxide

The Cauchy problem about Dirac-wave map from the 2-dimension Minkowski space to a complete Riemannian manifold

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Science china Mathematics 2007年第6期50卷 859-874页

作者： Yan ZHANG school of Statistics and Mathematics , zhejiang Gongshang university, hangzhou 310018, china department of Physics, zhejiang university, hangzhou 310027, china school of Statistics and Mathematics Zhejiang Gongshang University Hangzhou China department of Physics Zhejiang University Hangzhou China

When a target manifold is complete with a bounded curvature, we prove that there exists a unique global solution which satisfies the Euler-lagrange equation of for the given Cauchy data.

关键词： Dirac-wave map nonlinear σ-model spin bundle harmonic map 53C43

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